1.Effect of goat testis extract on nitric oxide synthase activity in injured Sertoli cells of mice
Hongyan TIAN ; Zhixin LI ; Ye XU ; Xinwei ZHU ; Yue ZHONG ; Moran CHEN ; Zhaohua DOU
Chinese Journal of Tissue Engineering Research 2006;10(47):215-217
BACKGROUND: Considerable studies demonstrate that nitric oxide synthase (NOS)/nitric oxide (NO)plays an important role in maintaining normal function of Sertoli cells, and influences spermatic generation and activation as well as fertilizability.OBJECTIVE: To observe the effect of goat testis extract on NOS activity in Sertoli cells of mice with testis injury caused by heavy mental Pb.DESIGN: Randomized controlled animal trial.SETTING: Department of Histology and Embryology, Jilin Medical College.MATERIALS: This trial was carried out in the laboratory of Histology and Embryology, Jilin Medical College (Key laboratory of the general logistics department of P.L.A) during March 2004 to August 2005. Thirty healthy Kunming male mice were involved and randomized into 3 groups,with 10 in each group: control group, testis injury model group (model group) and goat testis extract-treated group (treatment group).METHODS: The mice in the model group and experimental group were daily administrated with 100 g/L lead acetate, 0.2 mL/(mouse·d), 5 times/wk within 2 weeks, then withdrawal for 1 week. Simultaneously, the mice in the treatment group were subcutaneously injected with goat testis extract at 0.5 mL/(mouse ·d). The mice in the control group were given redistilled water of the same dose as that in the treatment group. After being poisoned fully, the mice were fasted for 12 hours and weighted, finally sacrificed by decapitation. Bilateral testis were dissected, immediately weighted, fixed with formalin and sliced. The NOS changes in Sertoli cells of mice in each group were observed with reduced nicotinamide-adenine dinucleotide phophate-diaphorase(NADPH-d) histochemical method combined with microscope image.MAIN OUTCOME MEASURES: Body mass, bilateral testis mass and NOS absorbance (A) in Sertoli cells of mice in each group after contamination expiration.RESULTS: All the 30 mice were involved in the result analysis, without deletion. ①After contamination expiration, the body mass, bilateral testis mass and NOS A value in treatment group were significantly than those in the model group [(22.47±3.49) g vs. (19.13±3.46) g;(0.113±0.021 ) g vs.(0.089±0.017) g; 0.236±0.020 vs. 0.146±0.023, t=2.151-3.314,P < 0.05-0.01]; ② In the model group, NOS positive Sertoli cells swelled and degenerated; The morphology of NOS positive Sertoli cells in the treatment group was close to that in the control group.CONCLUSION: Goat testis extract can boost the NOS activity in Sertoli cells of mice with testis injury caused by heavy mental pliumbum and has some repairing and protective effect on testis injury, which can provide new thinking for treatment of male sterility.
2.Analgesic effect of polygonum bistorta L. water extract
Zhaoyi ZENG ; Min WANG ; Heyang YE ; Li ZHOU ; Qing ZHOU ; Jing ZENG
Chinese Journal of Tissue Engineering Research 2006;10(47):199-201
BACKGROUND: Bistort rhizome is also named as caoheche, which is characterized by clearing heat, relieving convulsion, regulating damp and reducing swelling. Additionally, its water extract is characterized by antiarrhythmia and central inhibition; however, analgesia should be studied further.OBJECTIVE: To observe analgesic effect of polygonum bistorta L. Water extract, and compare with morphine and amidazofen.DESIGN: Completely randomized digital table and randomized controlled animal study.SETTING: Department of Pharmacology, Gannan Medical College.MATERIALS: The experiment was carried out in the Laboratory of Scientific Center of Gannan Medical College from March to May 2004. ① A total of 150 healthy adult Kunming mice were used in the 4 independent experiments. ② Medicines: Polygonum bistorta L. Water extract (Department of Phytochemistry, Shenyang Pharmaceutical University; batch number: 2003061001); morphine hydrochloride solution (Shenyang First Pharmaceutical Factory; batch number: 000305); naloxone hydrochloride solution (Yanqiao pharmaceutical Co. Ltd.; batch number: 20021109).METHODS: ① Effect of polygonum bistorta L. Water extract on twisting-body reaction of mice induced by acetic acid: Forty mice were randomly divided into 4 groups: saline group, 0.10 and 0.15 mg/g polygonum bistorta L. Water extract groups and amidazofen group with 10 in each group. All mice were intraperitoneally injected with 0.02 mL/g saline,0.10 and 0.15 mg/g polygonum bistorta L. Water extract solution and 0.10 mg/g amidazofen, respectively. Fifteen minutes later, mice were intraperitoneally injected with 6 g/L 0.01 mL/g acetic acid glacial to record times of twisting-body reaction within 15 minutes. ② Effect of polygonum bistorta L. Water extract on pain response of mice induced by hot-plate test: Forty female mice were randomly divided into 4 groups:saline group, 0.10 and 0.15 polygonum bistorta L. Water extract groups and morphine group with 10 in each group. All mice were intraperitoneally injected with 0.02 mL/g saline, 0.10 and 0.15 mg/g polygonum bistorta L. Water extract solution and 0.01 mg/g morphine solution, respectively. GJ-8402 hot-plate pain response threshold detector was used in this study; pain response temperature was (55.0±0.5) ℃; pain response after licking hindfoot was regarded as reactive marker; latency of pain response threshold was within 60 s. Pain response was measured at 15, 30 and 60 minutes after administration with hot-plate test. ③ Effect of morphine, naloxone and polygonum bistorta L. Water extract on pain response of mice induced by hot-plate test: Thirty female mice were randomly divided into 3 groups: saline group, naloxone+morphine group and naloxone+polygonum bistorta L. Water extract group with 10 in each group. All mice were intraperitoneally injected with 0.02 mL/g saline, 0.004 mg/g naloxone solution+0.01 mg/g morphine solution and 0.004 mg/g naloxone solution+0.15 mg/g polygonum bistorta L. Water extract solution, respectivelu. Pain response was measured at 15, 30 and 60 minutes after administration with hot-plate test. ④ Effect of polygonum bistorta L. Water extract on pain response of mice induced by electric stimulation: Forty mice were randomly divided into 4 groups with 10 in each group. All mice were intraperitoneally injected with 0.02 mL/g saline, 0.10 and 0.15 mg/g polygonum bistorta L. Water extract and 1 g/L morphine, respectively. Pain response was measured at 20, 35, 50 and 70 minutes after administration with electric stimulus method.MAIN OUTCOME MEASURES: ① Times of twisting-body reaction; ②duration of pain response induced by hot-plate test; ③ analgesic rate induced by electric stimulation.RESULTS: All 150 healthy adult Kunming mice were involved in the final analysis. ① Times of twisting-body reaction: At 15 inutes after administration, times of twisting-body reaction were less in 0.10 and 0.15 mg/g polygonum bistorta L. Water extract group and amidazofen group than those in saline group [(15.1±11.1), (8.0±6.5), (6.3±3.2), (54.1±20.2) times, t=3.532-3.681, P < 0.01]. ② Duration of pain response induced by hot-plate test:At 15, 30 and 60 minutes after administration, durations of pain response induced by hot-plate test were longer in 0.10 and 0.15 mg/g polygonum bistorta L. Water extract group and morphine group than those in saline group (t=2.676-3.683, P < 0.05-0.01). ③ Duration of pain response was longer in naloxone + polygonum bistorta L. Water extract group than that in saline group at each time point after administration (t=2.676-3.563, P< 0.05-0.01); however, duration in naloxone + morphine group was close to that in saline group (P > 0.05). ④ Analgesic rate induced by electric stimulation: At 20, 35, 50 and 70 minutes after administration, analgesic rate induced by electric stimulation was higher in 0.10 and 0.15 mg/g polygonum bistorta L. Water extract group and morphine group than that in saline group (t=3.455-3.634, P < 0.01).CONCLUSION: ① Polygonum bistorta L. Water extract has the obviously analgesic effect, whose intensity is close to that of amidazofen and morphine. ② Naloxone, an opiate receptor antagonist, can resist analgesic effect of morphine but not that of polygonum bistorta L. Water extract. This suggests that analgesic effect of polygonum bistorta L. Water extract dose not react through exciting opiate receptor.
3.Effect of Lujiao prescription on the degree of left ventricular hypertrophy in patients with congestive heart failure
Hui CAI ; Wanying HU ; Yanjun WANG ; Junhao GUO ; Bing HU ; Junhui ZHANG
Chinese Journal of Tissue Engineering Research 2006;10(43):222-224
BACKGROUND: Left ventricular hypertrophy (LVH) is an important ring-joint of congestive heart failure (CHF). It is also considered to be independent risk factor of cardiovascular disease, so how to reverse or relieve LVH is valuable to cure CHF.OBJECTIVE: To observe the effect of Lujiao prescription on degree of LVH in patients with CHF.DESIGN: A random-control observation on CHF patients.SETTING: Department of Cardiovasology, Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine.PARTICIPANTS: A total of 20 patients with CHF were selected from the Special Clinic and Department of Cardiovasology in Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from January 1996 to December 1998. All of them were consistent with the Framirham diagnostic criteria and following standardizations: ①above Ⅲ degree of cardiac functional grading by New York Heart Association (NYHA);②over 3 months case history;③accepted basic treatment of quantity sufficient diuretic and vasodilating agent excluding angiotensin converting enzyme inhibitor (ACEI);LVH was proved by echocardiogram or chest X-ray. They were informed of treatment items and consented.METHODS: According to random lot, the patients were divided into Lujiao prescription group and Digoxin group, each containing 10 patients.①Oral administrations of Lujiao prescription (35 mL, two times per day, produced by Shanghai Shuguang Hospital) and Digoxin (0.125-0.25 mg per tablet, batch number: 970757, once daily) were given in the corresponding groups, and the treatment duration was 3 months. After treatment, all the patients were detected in①NYHA cardiac functional grading; ②interventricular septal thickness (IVST) and posterior wall thickness (PWT) in diastolic phase through echocardiogram, myocardium weight of left ventricular by Devereux formula, left ventricular mass index (LVMI) corrected by bodysurface area, and degree of LVH;③the level of plasma angiotensin Ⅱ (Ang Ⅱ ) by radioactive immunoassay.MAIN OUTCOME MEASURES: NYHA cardiac functional grading, e-chocardiogram (IVST, PWT, LVMI, degree of LVH), the level of plasma Ang Ⅱ.RFSULTS: All 20 adopted patients entered the result analysis.①The classification of NYHA cardiac functional grading ( Ⅰ , Ⅱ ,Ⅲ,Ⅳ) were respectively 2, 5, 3, 0 case in Lujiao prescription group and 2, 4, 4, 0 case in Digoxin group after treatment, and difference was significant compared with those before treatment (0, 0, 7, 3, P < 0.05).②IVST, PWT and LVMI of Lujiao prescription group were obviously decreased after treatment [(11.20±0.42), (12.10±0.32) mm, P < 0.01; (10.60±0.84), (11.40±1.10) mm,P < 0.01; (139.4±12.4), (155.3±15.4) g/m2, P < 0.01], while those of the Digoxin group were untouched (P > 0.05). The degree of LVH of Lujiao prescription group after treatment was obviously decreased [normal, mild,moderate, sever: 2, 6, 2, 0; 0, 3, 4, 3, P < 0.01], while that of the Digoxin group was not obviously decreased (P > 0.05).③)The level of plasma Ang Ⅱin the Lujiao prescription group was obviously decreased [(97.7±19.5),(144.0±18.5) ng/L, P < 0.01], while that of the Digoxin group was not obviously increased (P > 0.05).CONCLUSION: Lujiao prescription can reverse or relieve the degree of LVH in patients with CHF.
4.Inhibitory effects of total flavones of buckwheat flower on the non-enzymatic glycation of proteins in vivo and in vitro
Fengling NIU ; Jinxiu CHU ; Shuying HAN
Chinese Journal of Tissue Engineering Research 2006;10(43):210-213
BACKGROUND: Non-enzymatic glycation of proteins is involved in the complications of diabetes mellitus. Previous experiments have demonstrated that total flavones of buckwheat flower (TFBF) could improve carbohydrate tolerance. However, it is little known whether TFBF inhibit the non-enzymatic glycation of proteins.OBJECTIVE: To investigate the influences of TFBF on the non-enzymatic advanced glycation end products (AGEs) of proteins in vivo and in vitro.DESIGN: Completely randomized controlled trial.SETTING: Department of Pharmacology, North China Coal Medical College.MATERIALS: Totally 75 adult SD rats , of clean grade, weighing (200±20) g, including 38 female rats and 37 male rats, were provided by the institute of experimental animals, Chinese Academy of Medical Sciences (Certification No. SCXK11-00-0006). TFBF was extracted by our laboratory from flowers of buckwheat. The blood glucose kit was purchased from Beijing Biosino Biotechnology Company Ltd. Penicillin (Batch No.031020, 8×105 U) and streptomycin (Batch No. 030920, 1×106 U) were purchased from North China Pharmaceutical Company. Streptozotocin and BSA were purchased from Sigma Company. Fructosamine kit was purchased from Nanjing Jiancheng Bioengineering Institute, and the other chemicals were analytical pure produced domestically.METHODS: This experiment was carried out in the Department of Pharmacology, North China Coal Medical College from March to October 2004.In the first experiment, in vivo macromolecular AGEs was measured: ①Modeling and grouping: Rats were divided into 3 groups according to body mass: Normal control group (n=15), the rats were treated with 8 mL/kg normal saline intraperitoneally. Streptozotocin-treated group (n=45), the rats were fasted for 16 hours and then treated with 80 mg/kg streptozotocin of 8 mL/kg intraperitoneally. Twenty-two hours later, the blood of all rats was harvested from vena caudalis to measure the level of blood sugar.Those with fasting blood glucose ≥ 15 mmol/L were acted as diabetic rats.Streptozotocin-treated group were divided into 3 subgroups, 15 rats in each subgroups. Each rat was given intragastric administration of 0.1, 0.2 and 0.4 g/kg TFBF. Model group (n=1S): Rats were only treated with 80 mg/kg streptozotocin of 8 mL/kg . The rats in normal control group and model group were given the same volume of salt water. The administration was once a day for 12 weeks successively. ②Measurement of fasting blood glucose: After the last administration, the rats of streptozotocin-treated group were fasted for 12 hours and the blood was harvested from vena caudalis. The fasting blood glucose was measured by glucose oxidase method. ③The levels of blood plasma and nephridial tissue fructosamine and macromolecular AGEs were measured: The rats of each group were anesthetized with ethyl ether on the second day following the last administration. Blood was chosen from carotid artery, and plasma was separated.Kidneys were taken at the same time, prepared into 100 g/L tissue homogenate and centrifuged at low temperature. The levels of fructosamine in plasma and the supernatant fluid of kidney homogenate were measured according to the instructions of the kit. AGEs in plasma and renal tissue were determined by fluorospectrophotometer. The products of macromolecular AGEs were calculated. In the second experiment, in vitro macromolecular AGEs were measured as below: 0.01, 0.05, 0.10 mg/L TFBF of 6 mLrespectively was prepared with solution A (0.2 mol/L glucose, 2×l06 U/Lpenicillin, 2×106 U/L streptomycin , PBS containing 20 g/L bovine serum albumin). Control groups were set: ① without TFBF, ② without TFBF and glucose, ③ without BSA, ④ without glucose. Five parallels of each sample were sterilized by filtration and incubated in the attemperator at 37 ℃. The fluorescence of AGEs (F) in the culture was determined at the 4th, 8th and 12th weeks. Inhibition ratio (IR) was calculated and the inhibition of TFBF on AGEs was observed.MAIN OUTCOME MEASURES: In the first experiment, the levels of fasting blood glucose, fructosamine in kidney and plasma, and AGEs were measured. In the second experiment, the inhibition of TFBF on AGEs in vitro was measured.RESULTS: In the first experiment, 75 rats were involved, and 56 successful rats entered the stage of result analysis. The levels of blood glucose,fructosamine in kidney and plasma of rats in the model group were significantly higher than those of rats in the normal control group (t=7.572,10.186, 5.794,P < 0.01 ). The level of blood glucose of rats in the 3 subgroups was significantly lower than that of rats in the model group (t=3.357,4.382,3.938,P < 0.05-0.01); The levels of fructosamine in kidney and plasma of rats in the 0.2 and 0.4 g/kg TFBF groups were significantly lower than those in the model group (t=5.109, 4.605, 3.731,3.097,P < 0.05-0.01 ). The levels of AGEs in plasma and kidney of rats in the model group were significantly higher than those in the normal control group (t=6.463, 12.704,P < 0.01 ), while the levels of AGEs in plasma of rats in the streptozotocin-treated group were similar to those in the model control group (P >. 0.05), and those in kidney of rats in the streptozotocintreated subgroups were significantly lower than those in the model group (t=9.845, 12.799, 12.899,P < 0.01 ). In the second experiment, the level of macromolecular AGEs of each group was gradually increased with ime.TFBFcould inhibit the formation of macromolecular AGEs in dose- and time-dependent manner.CONCLUSION: TFBF obviously inhibited the formation of AGEs of proteins in vivo and in vitro.
5.Expression of different genes in the lung tissue of model rats with chronic obstructive pulmonary disease: A comparison among normal, model control and Chinese herb groups
Wei ZHANG ; Jide JIN ; Xinyue ZHANG ; Yumeng SHAO
Chinese Journal of Tissue Engineering Research 2006;10(43):204-210
BACKGROUND: Many diseases accompany with changes of gene expression which can provide a valuable clue for pathogenesis and therapy of cognitive diseases.OBJECTIVE: To screen expression of different genes in lung tissue of model rats with chronic obstructive pulmonary disease (COPD) by gene expression profiling technique. DESIGN: Open study. SETTING: Affiliated Hospital of Shandong University of Traditional Chinese Medicine; Radiation Medical Institute of Academy of Military Medical Sciences of Chinese PLA; Shangdong University of Traditional Chinese Medicine.MATERIALS: The experiment was carried out in the Central Laboratory of Affiliated Hospital, Shandong University of Traditional Chinese Medicine from April to October 2005. ① Fifteen healthy male Wistar rats of SPF grade and aged 50 days were randomly divided into normal group, model control group and Chinese herb group with 5 in each group. ② Type of gene chip was BiostarR-40S and provided by Shanghai Boxing Gene Chip Company Limited (batch number: G050510010052).METHODS: ① COPD models in model control group and Chinese herb group were established with modified smoking-fumigated method plus adding lipopolysaccharide in windpipe. Pathological changes of airway were observed under optic microscope. Models were established successfully based on the following phenomenon: hyperemia and edema in mucous membrane of airway; degeneration and necrosis of epithelial cells; infiltration of bronchial cavity in lung tissue and surrounding chronic inflammatory cells; proliferation of smooth muscle and fibrocyte surrounding dissepiment; thin and broken interval; amplifying confluence of pulmonary alveolus; thickness of vascular wall of arteriole; decrease of vaso-cavity; infiltration of surrounding inflammatory cells. ② At 30 days after modeling,0.65 g/mL self-made Chinese herb (mahuang, xingren, huangqi, etc.) wasperfused into rats in Chinese herb group with 2 mL each time, once a day,for successive 14 days. Rats in model control group were perfused with 2 mLsaline; however, rats in normal group were untouched. All rats drank freely under the same internal environment. ③ After administration, rats in each group were sacrificed to extract total RNA, label and hybridizated with probe, wash pieces, analyze gray value of clip with clip imaging software,obtain original signal of each gene point (signal values of foreground and background) and determine differently expressed genes. If size of test was more than 2.0, genes were regarded as up-regulation genes; otherwise, if ratio was less than 0.5, they were regarded as down-regulation genes. In this study, based on reliability, if size of test was more than 2.5, they were regarded as up-relation genes; however, if ratio was less than 0.375, they were regarded as down-regulation genes.MAIN OUTCOME MEASURES: ① Comparisons of differently expressed genes in normal and model control groups; ② Comparisons of differently expressed genes in normal and Chinese herb groups; ③Comparisons of differently expressed genes in model control and Chinese herb groups.RESULTS: Fifteen rats were all involved in the final analysis. ① As compared with those in normal group, there were 57 differently expressed genes in model control group, involving in immunity, metabolism, signal transduction, adjusting of gene expression, cell cycle, cell transport, cell migration, fibrosis, etc. ② Gene expression in Chinese herb group reached normal level and there were 11 differently expressed genes as compared with those in normal group, involving in stress, signal transduction, cytoskeleton, etc. ③ As compared with those in model control group, there were 7 differently expressed genes in Chinese herb group, involving in immunity and excretion of neurotransmitter.CONCLUSION: Changes of gene expression may be one of reasons of COPD pathogeneses; moreover, most differently expressed genes can recover the normal level after drug therapy which is beneficial to correct pathological changes of COPD.
6.Effects of polyglycosides of tripterygium wilfordii hook F on the relative data of ankle joints of model rats with adjuvant arthritis
Qin YANG ; Guizhi YANG ; Lei WANG ; Yuqiong MA ; Hanwen TIAN
Chinese Journal of Tissue Engineering Research 2006;10(47):189-192
BACKGROUND: Reumatoid arthritis (RA) is a well-known autoimmune disease. Recently, polyglycosides of tripterygium wilfordii hook F (T Ⅱ), a traditional Chinese herb, has been widely used to treat rheumatoid arthritis.But the effects of TⅡ on the joints' synovium inflammation and whether TⅡcan prevent the reumatoid arthritis need to be investigated further.OBJECTIVE: To study the effects of T Ⅱ on the relative data of ankle joints of adjuvant arthritis (AA) rats.DESIGN: A randomized controlled experiment based on rats. SETTING: Departments of Histology and Embryology and Neurobiology, West China School of Preclinical and Forensic Medicine, Sichuan University. MATERIALS: A total of 20 healthy clean grade female SD rats, aged 2 to 3 months old, weighing 185-215 g, were provided by the Experimental Animal Center of West China Medical Center of Sichuan University. Fre und's complete adjuvant (FCA) was produced by Sigma Company. TⅡ was produced by Zhuzhou 3rd pharmacy of Hunan Province (certification number: 2005 No 055172, 10 mg/pill).METHODS: The experiment was completed in Department of Histology and Embryology and Neurobiology in Sichuan University from May 2004 to March 2005. ① All the 20 rats were randomly divided into 4 groups with 5 rats in each group by lots: normal group, without Freund's complete adjuvant (FCA) injection and TⅡ administration; model group, with FCA intradermal injection (0.2 mL) into the left hind paw and without TⅡ administration; TⅡ preventive group, first we use the same way as model group to replicate the AA model in rats, then on the 7th day AA rats were feed by TⅡ 30 mg/kg every day for 7 days; TⅡ therapeutic group, AA rats model were built with the same way as model group, on the 19th day AA rats were feed by TⅡ 30 mg/kg every day for 7 days. During this period, the swelling dimension of hind paw both primary and secondary are mea sured before immunization with FCA and after immunization, that was, on the 2nd, 10th, 15th, 19th, 22nd and 26th days. ② Arthritis index have been recorded according to inflammatory state of other three uninjected limbs.③ On the 28th day, all the rats were killed, the ankle joints are collected after perfusion-fixation. These joints were sectioned and colorated with H. E staining. Then we observe the histopathological changes in the synovium of ankle joints. MAIN OUTCOME MEASURES: Swelling dimension of joints and arthritis index, histopathology of anklebone joint's synovium. RESULTS: All of the 20 rats completed the experiment without missing. ① On the 2nd day after FCA injection, the primary hind paw of other three groups beside normal group appeared obvious swelling; from 2nd to 26th days, the volume of hind paw in other three groups was larger than that of normal group (t=2.315-3.041, P < 0.05). The volume of primary hind paw in TⅡ preventive group at different time points (10th, 15th, 19th, 22nd and 26th days) was obviously less than that of model group (t=2.064-2.683, P < 0.05). The volume of primary hind paw in TⅡ therapeutic group on the 22nd and 26th days was less than that of model group (t=2.112-2.578, P < 0.05). Fifteen days after FCA injection, the volume of secondary hind paw in model group and T Ⅱ therapeutic group was larger than that of normal group (t=2.201-2.546, P < 0.05). On the 15th, 19th, 22nd and 26th days, the volume of secondary hind paw in T Ⅱ preventive group was obviously less than that of model group (t=2.373-2.425, P < 0.05). The volume of secondary hind paw in T Ⅱ therapeutic group on the 26th day was obviously less than model group (P < 0.05). ② On the 14th day after FCA injection(after T Ⅱ preventive administration for 7 days), arthritis index of model group was (8.3±2.0) points, while arthritis index of TⅡ preventive group was (0.4±0.95) points (t=2.64, P < 0.05), there was an obvious decline in T Ⅱ preventive group compared with model group. On the 26th day after FCA injection (after TⅡ therapeutic administration for 7 days), arthritis index of model group was (11.2±0.7), whileinflammatory disease in AA rats and prevent the secondary arthritis in the rats of AA as well.
7.Immunosuppresive, antioxidative, anti-inflammatory and analgesic effect of Xiahuoluo pills
Jingqiang PAN ; Liuying XIAO ; Dan ZHANG ; Chao HAN ; Jieru LIN ; Linying ZHENG ; Boping LI
Chinese Journal of Tissue Engineering Research 2006;10(47):183-188
BACKGROUND: Xiaohuoluo pill can expel pathogenic wind, remove dampness and activate collaterals. It is used for treatment of Bi-syndrome due to wind-cold-dampness, pain and numbness in limbs.OBJECTIVE: To observe the pharmacological effect of Xiaohuoluo pills on secondary immune response, specific immunity (including cellular immunity and humoral immunity), non-specific immunity [including complement 3(C3), mononuclear phagocyte system (MPS) and red blood cell (RBC)adhesion function] and free radical injury as well as pain and many other inflammations in mice.DESIGN: A randomized controlled stratified trial.SETTING: Guangzhou Institute of Traditional Chinese Medicine and Chinese Materia Medica; Department of Pharmacy, Guangzhou Hospital of Traditional Chinese Medicine.MATERIALS: Totally 628 NIH and ICR mice of 6 to 8 weeks were involved in this trial. Xiaohuoluo pills (components: Dannanxing, Zhichuanwu, Zhicaowu, Dilong, Ruxiang and so on; Chenli Pharmaceutical Factory,Guangzhou; Brach No. 19980612) were used in this trial. Rabbit antimouse immunoglobulin G (IgG) and C3 antiserum reagent kit (Guangzhou Institute of Medicine and Health) and reagent kit for measuring the antioxidizing activity of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) (Jiancheng Institute of Bioengineering, Nanjing) were used.METHODS: This trial was carried out in the Guangzhou Institute of Traditional Chinese Medicine and Chinese Materia Medica; Department of Pharmacy, Guangzhou Institute of Medicine and Health during September 1998 to December 1999. ① To observe the suppressive effect of Xiaohuoluo pills on cock red blood cell (CRBC)-induced secondary immune response: Eight-four ICR mice, male and female in half, were selected.Twenty of 84 mice served as blank controls; The other 64 mice were intraperitoneally injected with cyclophos-phamide (CY) of 0.2 g/kgonce. On the 4th and 12th days, CRBC was intraperitoneally injected into the mice twice to induce immunoenhancing pathological models to form secondary immune response. Mice served as blank controls were intraperitoneally injected with the same volume of normal saline; The immunoenhanced mice were assigned into 3 groups by a lot: CY group (n=20, CY, 40 mg/kg, intragastric administration, I.g.), Xiaohuoluo pills (n=21, Xiaohuoluo pills suspension, 5.54 g/kg, I.g.) and model group (n=20, distilled water, the same volume as other groups, I.g.); once a day within 7 successive days. 19 days later, the levels of serum IgG and C3 were measured with single immunodiffusion method, and the level of circulating immune compound (CIC) was measured with polyethylene glycol precipitation method. ② To observe the suppressive effect of Xiaohuoluo pills on delayed type hypersensitivity (DTH): Fifty-four ICR mice, male and female in half, were selected. On the 1st day, the mice were sensitized by subcutaneous injection of 10 g/L 2,4-dinitrofluorobenzen e (DNFB) of 50 μL for each. On the 4th day, the sensitized mice were assigned into 3 groups by a lot: Prednisone group (n=18, prednisone, 0.01 g/kg, I.g.), Xiaohuoluo pills (n=18, Xiaohuoluo pills suspension, 5.54 g/kg, I.g.), model group (n=18, distilled water, the same volume as other groups, I.g.), all once a day within 7 successive days. 11 days later, 10 g/L DNFB of 25μL was spread on the right ear of each mouse in each group. The swelling degree was calculated 24 hours later (The mass difference between right ear and left ear). ③ To observe the suppressive effect of Xiaohuoluo pills on immune adhesion function of RBC of mouse: Thirty-six NIH mice, male and female in half, were selected and assigned into 3 groups by a lot: CY group (n=12, CY, 20 mg/kg,I.g.), Xiaohuoluo pills (n=12, Xiaohuoluo pills suspension, 5.54 g/kg, I.g.)and blank control group (n=12, distilled water, the same volume as other groups, I.g.), once a day within 7 successive days. 7 days later, blood was taken from the orbit of mice for calculating the rosette rate of RBC-C3b receptor and the rosette rate of RBC immune compound. ④ To observe the suppressive effect o20 Mg/kg, I.g.),Xiaohuoluo pills group (Xiaohuoluo pills suspension, 5.54 g/kg, I.g.) , once a day within 7 successive days; IgM-type hemolytic concentration (HC50)was measured at 2 hours after the last administration on the 7th day [ (Sample absorption / Absorption at HC50 of CRBC) ×diluted time]. The levels of serum C3 and MDA and the activity of SOD were measured according to the method from corresponding reagent kit. ⑥ To observe the suppressive effect of Xiaohuoluo pills on agar granulation tissue hyperplasia in mice:Fifty-nine NIH mice were selected and given subcutaneous injection of 20 g/L agar of 0.5 mL for each. 24 hours later, the mice were assigned into 3 groups by a lot: diclofenac group (diclofenac, 10 mg/kg,I.g..), Xiaohuoluo pills group (Xiaohuoluo pills suspension, 5.54 g/kg, I.g.) and model group (distilled water, the same volume as other groups, I.g.), once a day within 7 successive days; On the 8th day, the mice were sacrificed. The hyperplasiainhibiting effect was presented in the form of the mass of agar granulation tissue in one kilogram body mass ⑦ To observe the suppressive effect of Xiaohuoluo pills on acetic distortion reaction: Sixty-three NIH mice were se lected and assigned into 3 groups by a lot: diclofenac group (diclofenac,50 mg/kg, I.g.), Xiaohuoluo pills group (Xiaohuoluo pills suspension, 5.54 g/kg,I.g.) and model group (distilled water, the same volume of other groups, I.g.),once a day within 2 successive days. At 2 hours after the last administration, the mice were given intraperitoneal injection of 0.1 mol/L acetic acid of 0.2 mL for each one. The times of distortion of mice within 20 minutes were counted. ⑧ To observe the effect of Xiaohuoluo pills on the acute exudative inflammation evoked by dimethylbenzene, croton oil and carrageenan, and the level of prostaglandin E in the inflammatory exudates:Totally 219 NIH mice were selected and assigned into 3 groups by a lot:diclofenac group (diclofenac, 50 mg/kg, I.g.), Xiaohuoluo pills group (Xiaohuoluo pills suspension, 5.54 g/kg, I.g.) and model group (distilled water, the same volume of other groups, I.g.) once a day within 2 successive days. At 2 hours after the last administration, dimethylbenzene of 25 μL was spread on the right ear for 20 minutes, or croton oil of 25 μL was also spread on the right ear, 4 hours later, the swelling of right ear was calculated (mass of right ear-mass of left ear). 10 g/L carrageenan of 20 μL was subcutaneously injected into the right foot, 3 hours later, the swelling degree was calculated (The difference of right foot and left foot); and the level of prostaglandin E in the inflammatory exudates was measured. ⑨ t test(t' test for heteroscedasticity) was used for comparing the difference in measurement data among groups.MAIN OUTCOME MEASURES: Pharmacological effect of Xiaohuoluo pills on secondary immune response, specific immunity, non-specific immunity and free radical injury as well as pain and many other inflammations in mice.RESULTS: Totally 628 NIH and ICR mice were involved in result analysis. ① The level of IgG and CIC of mice in the model group was significantly higher than that in the other 3 groups respectively (P < 0.01),while the level of C3 was significantly lower than that in the other 3 groups (P < 0.05 to 0.01). ② The swelling degree of mice in the diclofenac group and Xiaohuoluo pills group was significantly lower than that in the blank control group respectively ( both P < 0.01). ③ The rosette rate of RBC-C3b receptor and RBC immune compound in the blank control group was significantly higher than that in the other 2 gro ups respectively (P < 0.01). ④ The phagocytic index (K value )in the diclofenac group and Xiaohuoluo pills group was significantly lower than that in the blank control group, respectively (both P < 0.01).⑤ IgM-type HC50 and the level of serum MDA of CY group and Xiaohuoluo pills group were obviously lower than those in the immune control group (P < 0.01),while the level of C3 was higher than that of immune control group, there was no significant difference in the activity of serum SOD between CY group or Xiaohuoluo pills group and immune control group (P > 0.05). ⑥The ratio of agar granulation tissue mass to body mass in the diclofenac group or Xiaohuoluo pills group was significantly lower than that in the model group(P < 0.01).⑦ The times of distortion of mice within 20 minutes in the diclofenac group or Xiaohuoluo pills group were signifi cantly less than those of model group(P < 0.01,0.05).⑧The ear swelling degree of dimethylbenzene-induced inflammatory models and croton oil-induced inflammatory models,and foot swelling degree of carrageenan-induced acute inflammatory models as well as the level of prostaglandin E in the inflammatory exudates in the diclofenac group were significantly milder or lower than those in the model group(P < 0.05 to 0.01),and the level of prostaglandin E in the inflammatory exudates in the Xiaohuoluo pills group was significantly lower than that in the model group (P < 0.01).CONCLUSION: Xiaohuoluo pills possess pharmacological effects of immunosuppression, anti-proliferative inflammation, analgesia and antioxidation.
8.Effect of clozapine on human sperm chromosomes in vitro
Chinese Journal of Tissue Engineering Research 2006;10(46):218-219
BACKGROUND: Clozapine is a common antipsychotic drug for treating psychotic patients. Some reports suggest that it can cause chromosomal aberration of human cells. This study was designed to analyze the effect on mutagenesis of human generative cells and genetics toxicity of generative cells.OBJECTIVE: To research the effect of clozapine on human sperm chromosome with testing system ex vivo.DESIGN: Randomized controlled observation on the basis of human sperm chromosome.SETTING: Center of Psychological health, the First Hospital affiliated to Chongqing Medical University.MATERIALS: Human sperm was selected from healthy adult males who were not received mutagenesis factors within half a year. Clozapine was provided by the Ninth Pharmaceutical Factory of Shanghai. Ovum was selected from female golden shrewmouse aged 6-8 weeks. Ovum was fertilized and washed with BWW culture medium containing 0.3% or 3.5% human serum albumin. After fertilization, ovum was cultured with ovi-culture medium containing 10% serum of shrewmouse.METHODS: At three days before experiment, shrewmouse was muscularly injected with 40 unit/ampoule preganat mares esrum gonadotrophin, and then with 30 unit/ampoule human chorionic gonadotrophin. Semen was maintained in aseptic beaker and made 5 mL sperm suspension after washing, centrifugation and capacitation. The suspension was equally put into 5centrifuged tubes. 40 mg/L bleomycin A5 was added into one tube to regard as positive control, one tube was regarded as negative control without adding any reagent, and other tubes were added with clozapine at the concentrations of 200, 400 and 800 μg/L, respectively. Ovarium mound cells and pellucid zone in ovum were wiped out with 0. 1% alidase and pancreatin, and then, equally transplanted into a blank gutta in five culture medium. Sperm chromosome was established with stepped fixed air technique.MAIN OUTCOME MEASURES: Sperm rate and broken amount of chromosomal structural aberration.RESULTS: When concentration of clozapine was at 200, 400 and 800 μg/L,respectively, sperm rate and broken amount of chromosomal structural aberration were not significantly different from those in blank control group, and there was also no significant difference among three concentration groups. However, there was significant difference between 40 mg/L bleomycin A5 group and negative control group (P < 0.01).CONCLUSION: Clozapine cannot damage human sperm chromosome through detecting the effect of mutagen on breakage of chromosome with testing system ex vivo, but it has other genetics toxic mechanisms on human sperm chromosome.
9.Clinical characters, pathogenesis and influencing factors of different tremors
Lijun SUN ; Ping GU ; Mingwei WANG
Chinese Journal of Tissue Engineering Research 2006;10(46):226-228
OBJECTIVE: To summarize and analyze the characters, pathogenesis and influencing factors of various tremors, so as to provide evidence for the identification, prevention and treatment of various tremors in clinic.DATA SOURCES: A computer-based online search of Medline database was undertaken to identify articles about tremor published in English between January 1998 and May 2005 with the keyword of "tremor". Meanwhile, Chinese relevant articles published between January 1998 and May 2005 were searched with computer in Chinese full-text journal net by using the keyword of "tremor" in Chinese. STUDY SELECTION: The data were primarily checked. Inclusive criteria: ① articles about the classification, etiological factors and influencing factors of tremor; ② retrospective investigation on specific events. Exclusive criteria: repetitive studies.DATA EXTRACTION: Totally 48 articles were collected, and 31 repetitive studies were excluded. Of the other 17 ones accorded with the inclusive criteria, 9 were the investigations about tremor, and 8 were the specific cases of tremor.DATA SYNTHESIS: Tremor is classified mainly according to its manifestations and the etiological factors of basic disease. Mechanical tremor,reflexion of central nervous system and central oscillator are the pathogenesis of tremor. According to the manifestations, tremor canbe classified into static tremor, kinetic tremor, essential tremor, postural tremor, unfixed tremor. According to the different etiological factors, tremor can be classified into enhanced physiological tremor, essential tremor syndrome (classical essential tremor, orthostatic tremor, task-specific tremor, undetermined tremor), dystonic tremor, cerebellar tremor, Holmes tremor, peripheral neuropathic tremor, drug-induced and toxic tremor and psychogenic tremor,and different treatments should be adopted according to different etiological factors in clinic.CONCLUSION: Tremor has similar manifestations, but the pathogenesis are different, so different treatments should be adopted according to different etiological factors in clinic.
10.Standard and evaluation of mental health in undergraduates
Chinese Journal of Tissue Engineering Research 2006;10(46):223-225
OBJECTIVE: Recent studies of mental health status in undergraduates are reviewed by literatures in mental health standard, measurement and influential factors. It is aimed to analyze the current status of mental health in undergraduates and provide references for related research of mental health, psychological counseling and psychological health in undergraduates. DATA SOURCES: The China Journal Full-text Database (CJFD) and Wanfang database were undertaken to id entify related articles published from January 1995 to August 2005 with the key words of "psyche of undergraduate, mental health, health standard" in Chinese. References of retrieved articles were searched. Mental health related books were retrieved such as Health Psychology, Psychic Health Measurement and so on.STUDY SELECTION: The data were selected primarily, and related articles of mental health standard, measurement and influential factors were included. Studies unrelated to undergraduates were screened out, and fulltexts of the other literatures were looked up.DATA EXTRACTION: A total of 40 articles of mental health standard,measurement and influential factors in undergraduates were selected, of which 17 articles accorded with topic and content were collected. There were 4 articles of mental health standard, 6 literatures (listing 6 types and 20 kinds of questionnaires and tests) of measurement and 7 literatures (6articles) of influential factors.DATA SYNTHESIS: Main content of mental health was intellective health, emotional health and spiritual health. ①No mental health standard was authorized and generally accepted in internal and external studies at present. The four signs of mental health designed by World Psychiatric Association (WPA) were praised and identified, which were harmony of body,intelligence and emotion; adaptation of the environment and modesty in each other; well-being; bringing ones ability into full play and living effective living. Mental health standard of Chinese undergraduates was composed of normal cognition, good emotion, perfect personality, coordinated interpersonal relation and appropriate self-consciousness. ②Present commonly used-mental health measurement for undergraduates was various, but there still was some disadvantage. Big Five Questionnaire (BFQ) and University Personality Inventory (UPI) were commonly used at abroad. Symptom Checklist-90 (SCL-90) was commonly used at home. ③Major factors influencing mental health of undergraduates were family, environment, age,sex, school, subject, religion and so on, and speciality had obvious effect on mental health of undergraduates.CONCLUSION: The importance of mental health of undergraduates is realized gradually by scholars, and more and more scholars at home and abroad study the mental health of undergraduates. How to design exact mental health standard and precise mental health measurement for undergraduates deserve further investigation. Meanwhile, it is necessary to deeply analyze the correlation between speciality and mental health of undergraduates.Fan BB, Zhang CH.Standard and evaluation of mental health in undergraduates.