Objective To evaluate the treatment effects on schistosomiasis hepatic fibrosis with Dangguibuxue decoction.Methods Thirty-four rabbits weighing 2.0-2.5 kg were selected.The procedure for the model of schistosomiasis liver fibrosis was as follow.Thirty-four snails infected by schistosoma japonicum were placed in triangle flask to escape cercariain.Abdominal skin of each rabbit wag infected by 100 schistosoma japonicum cercariaes.All rabbits were housed in conventional cages at room temperature.After 13 weeks schistosoma japonicum infected,the rabbits'livers were examined by B-ultrasound examination every week.After 18 weeks of infection,B-ultrasound showed images of schistosomiasis liver fibrosis,the rabbits were thus identified as schistosomiasis liver fibrosis.Praziquantel Was as insecticide to treat rabbits.All rabbits were divided into treatment and control groups according to body weigh.The high and low dosage group were treated with Dangguibuxue decoction at 50 g/kg and 5 g/kg(n=11),respectively,and the control group with no treatment(n=12).Two groups were treated with Dangguibuxue decoction every day for 10 weeks.After 10 weeks of treatment,the rabbits were killed and the livers were obtained to go HE staining and histopathological examination under microscope.Results After 18 weeks of infection,B-ultrasound showed that the liver parenchyma Was coarse and net-shaped.having hyperechoie spots with lightbrand and facula.The liver's shape Was abnormal,portal and splenic vein diameter was widened.Histopathological examination showed that the high dosage group presented fibrosis,which was formed by collagen fiber extending to peripherM regions,low dosage group showed mild fibrosis,mild collagen fiber present with extension without compartment formation, while the control group presented severe fibrosis and severe collagen fiber present with a thickening of the partial compartments and the distribution of eggs nodules. The liver histopathological ranking results from different groups showed that the difference was significant (χ2= 13.585,P < 0.01), the difference between the treatment groups of high and low dosage and control group was significant,respectively(all P < 0.01 ). Conclusions The histopathology results showed that both high and low dosage Dangguibuxue decoction can reverse or eliminate schistosomiasis liver fibrosis in rabbits.

AIM: To study mitochondrial mass and structural protein changes in dexamethasone (DEX)-mediated mouse thymocyte apoptosis process. METHODS: DEX-induced mouse thymocyte apoptosis model was established. Annexin V-FITC/PI double staining was used to identify apoptotic and necrotic cells by flowcytometry, JC-1 staining was adopted to test mitochondrial membrane potential (△?_m), and cellular structural protein changes were studied with CFDA-SE staining. RESULTS: By 1?10~(-6) mol/L DEX stimulation, the apoptotic rate was 51.25%?5.51% and had significantly difference from control group (12.03%?2.00%); the necrotic rate in DEX group was 30.25%?3.67% and also had significantly difference from control group (10.11%?1.11%, P

Objective To establish a HPLC/MS/MS method for the determinati on of gentiopicroside in human urine.Methods The urine sample was treated by solid-phase extraction with internal standard of caffeine;The RESCEK C8 colum n(150mm?2.1mm,5 ?m) was used as the analytical column with a mobilep hase consisting of methanol-10mmol? L-1 NH4AC buffer(pH=6.5)-acetonit rile(50∶40∶10,V/V),the flow rate was 0.2 mL? min-1;A triple quadruple tandem mass spectrometer was used as the detector,Electrospray ioniza tion source was applied and operated in positiveion mode.Gentiopicroside and caffeine were detected by monitoring the ion transition of m/z 374.1→ 195.2 and m/z 195.2→ 138.2 respectively.Results The linear range was 30～ 9000 ng?mL-1(r=0.9980) for gentiopicroside in human urine.The recovery was 91.10% ～ 9 6.21 %,The absolute recovery was 100.52% ～ 103.83%,The within-day and between-day precisions were less than 10 %.Conclusion The method is proved to be sensitive,accurate,rapid,specific.

Objective To prepare a high‐titer rabbit specific serum antibody against Kaposi′s sarcoma associated herpesvirus (KSHV) ORF65 capsid protein and identify the specificity of serum antibody .Methods Artificial synthetic peptide of ORF65 pro‐tein was emulsified with Freund adjuvant .4 rabbits were immunized with the prepared antigen by subcutaneous injection at various sites of skin of back and jaw once every two weeks .Immunization was carried out in total 4 times .The serum of the immunized rab‐bits was collected at a week after the last immunization .The titer of rabbit anti‐serum was assayed by ELISA .Specificity of the rab‐bit anti‐serum was analyzed by immunofluorescence assay and Western blot .Results The immunized rabbits produced high‐titer se‐rum antibody after total immunization .The highest titer of anti‐serum against ORF65 protein peptide was 1∶12 800 .The results of Immunofluorescence assay showed that antibody was binded in plasma of BCBL‐1 cell mostly ,which was consistent with the expres‐sion location of ORF65 in BCBL‐1 cell .Subsequently ,the data of Western blot revealed a specific band about 21 kD which accorded with the size of ORF65 protein .Meanwhile ,the expression of ORF65 in TPA treated BCBL‐1 cells was higher than the control cells ,which was consistent with the expression characteristics of lytic protein .Conclusion High‐titer specific rabbit serum antibody against KSHV capsid ORF65 antigen could be successfully prepared by rabbits immunization with ORF65 protein peptide .The pre‐pared antibody could be revealed immune reaction specificity with KSHV ORF65 protein .

Objective To screen the differentially expressed proteins of blood stasis blocking tendon and vessel syndrome and liver-kidney deficiency syndrome of the steroid-induced osteonecrosis of the femoral head ( SONFH) by proteomic technology, so as to supply evidence for Chinese medical syndrome classification. Methods The serum was taken separately from 10 patients with blood stasis blocking tendon and vessel syndrome, 10 patients with liver-kidney deficiency syndrome and 10 healthy volunteers. The two dimensional electrophoresis combined with mass spectrometric method was applied to screen and identify the differentially expressed proteins, and then the obtained protein candidates were verified by Western blotting method. Results Seven proteins were identified from differentially expressed protein spots, including hemoglobin subunit delta, actin, complement C4, antithrombin-Ⅲ, apolipoprotein A-IV, leucine-rich alpha-2 glycoprotein and serum amyloid A-2 protein. We found that antithrombin-Ⅲ and serum amyloid A-2 protein had specific expression in the SONFH patients with blood stasis blocking tendon and vessel, and complement C4 and leucine-rich alpha-2-glycoprotein had specific expression in the SONFH patients with liver-kidney deficiency syndrome. The results of Western blot method showed that the expression levels of complement C4 and antithrombin-Ⅲ were down-regulated in SONFH patients ( P<0.05 compared with the healthy volunteers) , the down-regulation of complement C4 was more obvious in SONFH patients with liver-kidney deficiency syndrome (P<0.05) and the down-regulation of antithrombin-Ⅲ was more obvious in the patients with blood stasis blocking tendon and vessel (P<0.05), the results being accorded with those of proteomic detection. Conclusion Antithrombin-Ⅲand serum amyloid A-2 protein may be the specific serum protein markers of SONFH patients with blood stasis blocking tendon and vessel syndrome, and complement C4 and leucine-rich alpha-2-glycoprotein may be the specific serum protein markers of SONFH patients with liver-kidney deficiency syndrome.

BACKGROUND:As a new self-synthetized medical orthopedic fixation material, micron traditional Chinese medicine functional lining material can promote the secretion of alkaline phosphatase and cal us formation, decrease serum calcium level, as wel as increase serum phosphorus level and calcium-phosphorus product. OBJECTIVE:To further verify the therapeutic effect of micron traditional Chinese medicine functional lining material on fracture healing. METHODS:Twenty-four New Zealand rabbits were selected to prepare models of right radial fracture, and then were randomly al ocated into four groups:rabbits in micron group were treated with micron traditional Chinese medicine functional lining material (micron Panax notoginseng powder mixed with active ingredients of Daval ia divaricata and Carthamus tinctoriousplus) plus plaster external fixation;those in traditional Chinese medicine control group treated with ordinary Chinese medicine functional material plus plaster external fixation;those in lining material control group treated with ordinary lining material (sterile degreasing cotton) plus plaster external fixation;those in model control group received no treatment. The whole blood viscosity at high/middle/low shear rates and plasma viscosity were detected at 2, 4 and 6 weeks before and after modeling. Additional y, rabbits in each group were kil ed at 6 weeks after modeling, and radial samples were removed for histological observation. RESULTS AND CONCLUSION:The whole blood viscosity at high/middle/low shear rates and plasma viscosity in the micron group were al lower than those in the other three groups in the same period, and had significant differences from those in the model control group at 4 and 6 weeks after modeling (P<0.05 or 0.01). A large number of osteoblasts could be found, woven bone began to form and the bone marrow cavity was recanalized in the micron group;in the traditional Chinese medicine group, woven bone, chondrocytes and endochondral ossification coexisted and there was a connection between cal uses inside and outside of the broken end, but the bone marrow cavity was not recanalized. In contrast, neither connection between cal uses inside and outside of the broken end, nor recanalized bone marrow cavity appeared in the lining material and model control groups. In conclusion, micron traditional Chinese medicine functional lining material can promote fracture healing by improving microcirculation and accelerating osteoblast proliferation.

Objective To explore the growth and developmental parameters and behavioral characteristics of rhesus monkeys during the first year of birth and to establish the background data.Methods A total of 18 (♂=11,♀=7 ) infant rhesus monkeys born from individually caged mothers and with known genetic background and postnatal days were monitored monthly for body weight, body height, head circumference, chest circumference, forelimb length, hind limb length, crown-rump length, tail length and anal-genital distance from postnatal day ( PND) 1 to 360, while hematology, blood chemistry and lymphocyte subsets were examined on PND 28, 175 and 360, and finger maze test was carried out on PND 208.Results The body weight showed linear growth with no significant difference between genders (P>0.05). Except for the anal-genital distance of male infants was significantly greater than that of female infants ( P<0.01 ) , no significant differences were observed between sexes in other morphological parameters.No significant differences of hematology were seen between genders (P >0.05).Compared with that at PND28, TP and BUN were significantly increased (P<0.01) while ALP decreased with no significant difference (P>0.05) at PND 175 and 360.Compared with that at PND28, CD4 +and CD4 +/CD8 +were significantly decreased ( P<0.01) while CD8+significantly increased ( P<0.01) at PND175 and 360.The number of sessions to solve task 2 in learning test was significantly greater than other tasks with females significantly less than males ( P<0.05) .The females had higher correct rate than males in the 2-day random memory test (P<0.05).Conclusions Body weight and morphological parameters show a linear growth.The PND.The RBC, HGB, LYMPH, TP, BUN, ALP, CD4 +, CD8 +and CD4 +/CD8+in hematology, blood chemistry and lymphocyte subsets show relevant changes to the growth and development of organs and systems in infants, which should be highly concerned in drug evaluation.The finger maze test indicates that female infants have better reversal learning and long-term memory than male infants.Background data and behavioral characteristics of infant rhesus monkeys during the first 12 months of birth are established in this study, which provide useful reference and support the evaluation of developmental and reproductive toxicity of drugs in rhesus monkeys.

Objective To observe the pathological changes of bone hydatid cyst of meriones meridianus after radiation therapy,and to investigate the clinical effect of radiotherapy on bone hydatid disease.Methods Ascus was dissected sterilely from sheep liver that naturally infected with Echinococcus granulomas,sheared and sac skin removed.Then it was washed and precipitated with 0.9％ sterile saline for 3 times,and scolex was HE stained and counted,from which a 20 ml suspension was made containing 12 × 106/L of scolex.Health meriones meridianus (referred to as gerbil) 140,male and female were in each half,aged 2 to 3 months,body weight(38 ± 6)g,were involved in the study.Gerbil was injected a 0.2 ml suspension containing Echinococcus granulomas scolex into hind tibial periosteum,and X-ray was taken 12 months after the injection.According to the bone destruction in the vaccination site,gerbil hindleg tibia with clear jagged bone destruction was treated as inclusion criteria,and 72 animal were selected as gerbil bone hydatid disease animal models,male and female were in each half.A tatal of 72 gerbils were randomly divided into 4 groups:control group,40 beequerel(Gy) group,50 Gy group and 60 Gy group,18 rats in each group,male and female in each half.The model animals were treated with radiotherapy for 5 times,with 2 d interval,and radiation dose was 300 cGy/min.Each group of gerbils was sacrificed after radiotherapy,bone Echinococcus granulomas cysts was taken out sterilely,and observed by light and electron microscope.Intracapsular cyst fluid was extracted,washed and precipitated with 0.9％ sterile saline repeatedly,and and the pellet was HE stained for observation of scolex morphology and activity by light microscope.Results The morphology and activity ofEchinococcus granulomas in cystic fluid in control group were normal; the morphology and activity of Echinococcus granulomas were still normal in the 40 Gy group,and Echinococcus granulomas was not stained red; but those were abnormal,deformation and atrophy and stained red in the 50 Gy group; and were stained red,deformed,fractured and were wrapped by unknown in the 60 Gy group.By light microscope,the germinal layer,cuticle layer,brood capsule and histological structure of protoscolex were basically normal in irradiated region in the control group.The pathological changes of hydatid cyst in the 40 Gy group were mainly degeneration,structure of hydatid cyst was abnormal,stratum corneum was extensive edema,germinal layer became thinner and the fertile cyst was rare.The main pathological change of hydatid cyst in the 50 Gy group was that corneous layer was widely fractured,and the germinal layer was edema,buckling folds,cells decreased,rare seen brood capsule and scolex; the main pathological changes of hydatid cyst were mainly necrosis in the 60 Gy group,cuticle was extensive fault,stratum corneum and germinal layer was separated,germinal layer was atrophy and disorder,no brood capsule and scolex.By electron microscope,cuticle structure of Echinococcus granulomas cyst was clear,microvillus arranged neatly,morphology and structure of the cell and organelle in cytoplasm were normal in the control group.There were many inflammatory cells infiltrating germinal layer of Echinococcus granulomas cyst,microfilament and contents in microfilament were reduced in the 40 Gy group.Microvillus of Echinococcus granulomas disappeared,nuclear membrane was unclear,endoplasmic and mitochon eclasis,lymphocyte nuclear chromatin was clumping and edge set and in circular permutation in the 50 Gy group.Microvillus disappeared,perinuclear membrane indistinct and ruptured,parts of nucleoli were fragmented and marinated,endoplasmic reticulum was extensive expansion,mitochondria was pyknosis and obvious vacuolization,lymphocyte nuclear chromatin clumping and edge set,lysosomes and macrophage emerge in the 60 Gy group.Conclusions Radiotherapy can destroy the morphology and structure of bone hydatid cyst,radioactivity at 50 Gy has a lethal effect on hydatid cyst.Radiation treatment of bone hydatid disease has a good clinical effect.

BACKGROUND: External preparations treatment of fractures in traditional Chinese medidne are mostly ordinary hard-paste, cream, ointment, which lack of modernized traditional Chinese medicine transdermal delivery systems, and the treatment has poor drug absorption and simple process. The treating procedure needs to be used repeatedly, which would affect fracture external fixation and lead to unsatisfactory therapeutic effect. In this Paler, the extemal treatment of traditional Chinese medidna was combined with advanced micro-nano-technology to develop a new medical complex functional materials-micron traditional Chinese medicine functional lining material, which is an organic combination of diaplasis, fixation and local drug remedy, which provides a better treatment for fractures. OBJECTIVE: To explore the effect of micron tradifional Chinese medicine functional lining material on the healing of fracture in rabbits, METHODS: After model preparation, rabbits were randomly divided into 4 groups: rabbits in the micron group were treated with micron traditional Chinese medicine functional lining material plus plaster external fixation; those in the traditional Chinese medidna control group were treated with ordinary Chinese medicine functional material plus plaster extemal fixation; those in the lining material control group were treated with ordinary lining material added plaster extemal fixation; those in the natural healing control group had no special treatments. The effect of fracture healing was evaluated by X-ray at 2, 4 and 6 weeks after modeling. The serum concentration of alkaline phosphatase, calcium and phosphorus was detected respectivaly before modeling and at 2, 4 and 6 weeks after modeling. RESULTS AND CONCLISION: ①X-rap examination:After 6 weeks the external was almost copletly absorbed and medullary cavity was completely recanalizad in the micron group, while fracture line disappeared and the external callus began to be absorbed and medullary cavity was not recanalized in the traditional Chinese medidne control group. Additionally, there were case of non-union stump hardening in natural healing group. ②Serum biochemistry:The activity of alkaline phosphatase reached a peak after 2 weeks in micron group, which was 2 weeks ahead of time compared with that in traditional Chinese medicine control group. And the peak concentration of alkaline phosphatase in micron group was higher than other groups. After 4 weeks serum calcium concentration started to fall significantly and phosphorus concentration started to rise significantly in micron group, as was statistically significant compared with other groups. After 6 weeks serum calcium concentration in micron group had fallen to normal level, while that in other groups was higher than normal. And phosphorus concentration in micron group increased significantly, which was statistically significant compared with natural healing group. The results demonstrated that micron feature traditional Chinese medicine lining matedat can enhance the activity of alkaline phosphatase, decrease the serum concentration of calcium and increase the serum concentration of phosphorus and the product of calcium and phosphorus, therefore, promote the healing of fracture, It shows that micron traditional Chinese medicine functional lining matedal plays a stronger effect on promoting the healing of fracture than that of ordinary Chinese medicine functional lining material.

OBJECTIVE:To prepare a three-pulse drug release system of nimodipine and study its drug release.METHODS:Solid dispersion technique and dry-coating method were respectively applied to prepare immediate-release mini-tablets and Pulsatile mini-tablets with lag-time of 4 h or 8 h.Then those mini-tablets were filled into capsule to obtain a three-pulse drug release system and subjected to in vitro dissolution test.DSC was employed to determine drug status in solid dispersion.RESULTS:Immediate-release mini-tablets were released more than 95% in 30 min.Pulsatile mini-tablets were released less than 10% in 4 h or 8 h of lag-time period.After lag-time period,pulsatile mini-tablets were released completely in 3 h.The whole pulsatile drug release system achieved three times of drug release at 5 min,4 h,8 h,respectively.Nimodipine kept amorphous form and were delivered into carrier evenly.CONCLUSION:A three-pulse drug release system of nimodipine has been prepared successfully.