AIM: To explore the relationship between the expression of caspase-2 and caspase-3 and the apoptosis in retinal ischemia/reperfusion (I/R) injury of rats, as well as the therapeutic effects of brain derived neurotrophic factor (BDNF)on the ischemic and reperfused retina.METHODS: This experiment was conducted at the laboratory of Affiliated Hospital of Qingdao University Medical College from February 2007 to July 2007. The models of retinal ischemia/reperfusion injury were made by transiently elevating intraocular pressure. A total of 28 rats were divided into Normal and Operative Groups. Operative group was divided into six subgroups. In each subgroup there were four rats. The left eyes of rats were used for I/R and the right eyes were used for intravitreal injection of brain-derived neurotrophic factor (BDNF) as treatment group. After reperfusion we divided our subgroups according to the reperfusion time as 1, 6, 12, 24, 48, 72 hours. The retinal ganglion cell number was counted by using optic microscope(BX-51,Olympus). Apoptosis was assessed by the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labelling (TUNEL) method, and the expression of caspase-2,caspase-3 was studied by enzyme linked immunosorbent assay (ELISA) and strept avidin-biotin complex (SABC)immunohistochemistry.RESULTS: No positive apoptotic cells were observed in the normal rats' retinae, but there were a significant number of positive apoptosis cells in 6-24 hours after transient ischemia followed by a decrease at 48 hours. The number of apoptotic cells reached a maximum at 24 hours after ischemia .The expression of caspase-2 gradually increased as early as at 6 hours, reached a peak at 24 hours, then decreased between 48 and 72 hours. Similarly, caspase-3 has the same rule with caspsae-2 in the time courses of expression in retinal tissues.BDNF administered before reperfusion inhibited the expression of apoptosis and ameliorated the retinal tissue damage. It also decreased caspase-2 and caspase-3 expression in ischemic/reperfused retina.CONCLUSION: Retinal ischemia-reperfusion can induce apoptosis of cells in the retina. BDNF rescues retinal ganglion cells (RGCs) from retinal ischemia/reperfusion injury through down-regulation of cell apoptosis and caspase-2 and caspase-3 expression. BDNF have a neuroprotective effect on retina.

OBJECTIVETo investigate the influence of cigarette smoking on human sperm DNA integrity.

METHODSTotally, 784 cases of male infertility were selected from our case database and grouped according to whether they were smokers or nonsmokers, how much they smoked (< or = 10, 11-19 and > or = 20 cigarettes/d) and how long they smoked (< or = 5, 6-9 and > or = 10 yr). Sperm DNA integrity was measured using sperm chromatin structure assay (SCSA) and flow cytometry. DNA fragmentation and immature spermatozoa were expressed by the DNA fragmentation index (DFI) and high DNA stainability (HDS) respectively. Conventional sperm parameters and sperm DNA integrity were compared among different groups.

RESULTSThe total semen volume and percentage of grade a + b sperm were lower and the sperm morphological abnormality was higher in the > or = 20 cigarettes/d and > or = 10 yr groups than in the others (P < 0.05). DFI and HDS were significantly higher in the smokers than in the nonsmokers (P < 0.05). HDS was negatively correlated with the percentage of grade a + b sperm (r = -0.18, P < 0.05) and both DFI and HDS were positively correlated with the rate of sperm malformation (r = 0.31 and r = 0.39, P < 0.05).

CONCLUSIONSmoking more than 20 cigarettes a day or longer than 10 years has deleterious effects on the semen volume, percentage of grade a + b sperm and sperm morphology of the smokers. Cigarette smoking decreases sperm DNA integrity and nuclear maturation.

Adult ; DNA Damage ; drug effects ; DNA Fragmentation ; Humans ; Infertility, Male ; genetics ; Male ; Middle Aged ; Smoking ; adverse effects ; Sperm Count ; Sperm Motility ; Spermatozoa ; drug effects ; Young Adult

Objective To study the effects of cold on the function of co lon in Sprague Dawley (SD) rats.Methods Eighty SD rats were randomly divided into normal group and experimental group.The rats in experimental group were orally administrated 0-4 ℃ cold saline for 60 days.The intake and drinking in 24 hours,the faeces quantity within 6 hours and the time of defecation were measured on 30 and 60 days.Blood biochemical parameters were measured with an automatic biochemical analyzer.Faeces were collected and observed by stereo microscope.A part of colonand gastric was fixed with 10％ for malin.Hematoxylin-eosin stained,and then observed under microscope.Results On 60 days,the weight of normal group and experimental group were (268.2 ± 18.35),(319.2 ± 10.17) g,and the intake in 24 hours were (48.0 ± 1.7),(85.1 ± 2.6) mg · g-1 and drinking in 24 hours were (54.6 ± 9.5),(84.6 ± 2.7) μL · g-1,the faeces quantity within 6 hours were (0.65±0.43),(3.06 ± 0.62) g,the time of defecation were (615.5±156.48),(408.2 ± 29.7) min.The difference between two groups above factors had statistical significance (all P ＜ 0.01).On 60 days,the serum Albumin (Alb) in experimental group was (38.48 ± 2.95) g · L-1,and the creatinine (Cr) was (36.46 ± 2.07) mmol · L-1,and the blood urea nitrogen (BUN) was (6.38 ± 0.46) mmol · L-1,and the concentration of sodium (Na +) was (142.04 ±4.43) mmol · L-1,and the concentration of potassium (K+) was (5.98 ± 1.03) mmol · L-1.There was no significant difference between two groups (all P ＞ 0.05).Observation of the morphology of colon that the mucosa epithelial cells and goblet cells in experimental group were increased,and the neutrophil were reduced and a small amount of inflammatory cell are infiltrating after 60 days.The rat feces become into beaded,hard and unpolished after 60 days.Conclusion Cold stimulation could lead to colonic motility and secretion dysfunction in SD rat.

Objective To study the effects of Angelica sinensis radix (Danggui) on gastrointestinal hormone and expresomatostatinion of tyrosine kinase receptor C-kit in mouse with blood-deficiency constipation (Xuexu Bianmi).Methods Kunming mouse were randomly divided into six groups,each group had 10 mouse:normal group,model group,control group,three dose of experimental groups.Except the normal group,the mouse in rest groups were orally administrated every day with diphenoxylate 25mg · kg-1 and subcutaneously injected at 1,4,7,10 d with acetylphenyhydrazine 60 mg · kg-1 and intraperitoneally injected at 10-14 d with cyclophosphamide 40 mg · kg-1 to copy Xuexu Bianmi model.At the 14 d,normal and model group were administered physiological saline,the control group were administered solution of Changtongshu Keli 5 mg · kg-1,three dose of experimental groups were administered with different doses of Danggui decoction (4.17,8.33,16.67 g · kg-1).The levels of motilin(MLT),substance P(SP),vasoactive intestinal peptide(VIP),cholecystokinin-8 (CCK-8) and somatostatin in blood were measured by enzyme-linked immunosorbent(ELISA).The expression of C-kit in colon were measured by immunohistochemical technique.Results MLT (pg · mL-1) in normal group,model group,control group,three dose of experimental groups were 1830 ± 246,1498 ± 198,1722 ± 237,1697 ± 245,1754 ± 199,1865 ± 259,respectively;the SP (ng · L-1) in the 6 groups were 266 ± 30,222 ± 34,259 ± 32,218 ± 28,252 ± 29,268 ± 30,respectively;the VIP (ng · L-1) in the 6 groups were 435 ± 109,579 ± 107,508 ± 105,417 ± 139,413 ± 126,445 ± 101,respectively;CCK-8 (pmol · L-1) in the 6 groups were 229 ±26,138 ± 12,180 ±22,164 ± 16,159 ±26,210 ±31,respectively;SS (n · L-1) in the 6 groups were 100 ±20,138 ±29,93 ± 16,104 ±23,101 ±22,99 ± 16,respectively.Comparison between model group and normal group,the di-fference had significantly (P ＜ 0.01,P ＜ 0.05).Comparison between control group on the CCK-8,SS and model group,the difference had significantly(all P ＜0.01).Comparison between experimental-L group on the VIP,CCK-8,SS and model group,the difference had significantly (all P ＜ 0.05)Comparison between experimental-M group on the MLT,VIP,SS and model group,the difference had significantly (all P ＜ 0.05).Comparison between experimental-H group on the MLT,SP,VIP,CCK-8,SS and model group,the difference had significantly(P ＜0.05,P ＜ 0.01).Comparison with the model group,the expression of C-kit in colon in three doses experimental groups were increased.Conclusion The mechanism of Danggui in treating mouse woth Xuexu Bianmi may be related to adjusted gastrointestinal hormone in serum and colon and increased the expression of C-kit in colon.

Objective To observe the effects of Angelica Sinensis Radix on the expression of aquaporin 8 (AQP8) and the levels of AC-cAMP-PKA in colon of blood-deficiency constipation in mice; To identify mechanism of Angelica Sinensis Radix for loosening the bowels to relieve constipation. Methods Sixty Kunming mice were randomly divided into control group, model group, positive medicine group, and Angelica Sinensis Radix high-, medium-, and low-dose groups, with ten mice in each group. Diphenoxylate, acetylphenlyhydrazine and cyclophosphamide were used to establish blood-deficiency constipation mice models. From the 14thday of the experiment, Angelica Sinensis Radix high-, medium-, and low-dose groups were given 16.7, 8.8, and 4.2 g/kg Angelica Sinensis Radix Decoction for gavage. Positive medicine group was given 5.0 g/kg Changtongshu Granules Liquid for gavage. Control group and model group were given equal volume of saline for gavage. The symptoms of blood deficiency constipation were observed and defecation time. Immunohistochemistry, Western blot and qRT-PCR were used to detect the expression of AQP8 protein and mRNA in colon, and the expression of AC-cAMP-PKA in colon was detected by ELISA. Results The mice in the model group developed blood deficiency constipation syndrome; the defecation time was significantly prolonged (P＜0.01); the expression level of colonic AQP8 protein and mRNA, AC, cAMP and PKA significantly increased (P＜0.05, P＜0.01). Compared with model group, the defecation time was significantly shortened in Angelica Sinensis Radix high-, medium-, and low-dose groups; the expression of AQP8 protein and mRNA and the levels of AC, cAMP and PKA were significantly decreased (P＜0.05, P＜0.01). Conclusion The treatment of Angelica Sinensis Radix for blood-deficiency constipation may be related to adjusting the AC-cAMP-PKA signaling pathways and reducing the expression of AQP8 protein and mRNA.

This study was aimed to observe the curative effect and safety of Danzhi Jiangtang Capsule ( DJC ) combined with atorvastatin on carotid artery intima-media thickness (IMT) in diabetes patients without hyper-tension . A total of 196 diabetes patients without hypertension with incrassate carotid artery IMT were randomly divided into the control group ( 98 cases ) and the treatment group ( 98 cases ) . The conventional diabetes thera-py was given to both groups . The atorvastatin of 20 mg/night was given to the control group . And the atorvas-tatin 20 mg/night added with DJC 9 . 0 g/night were given to the treatment group . The treatment course was
12 months . Carotid artery IMT , carotid atherosclerotic plaque area , FPG , FIns , HOMA-IR , HbA1c , blood lipids , hepatorenal function and etc . were examined before and after the treatment respectively . The results showed that there was a significant positive correlation between carotid artery IMT and FIns , HOMA-IR , HbAlc , LDL-C . After 12-month treatment , the total effectiveness is 85 . 87% in the treatment group . And there was significant difference compared with the control group ( P < 0 . 05 ) . The levels of FPG , FIns , HOMA-IR , HbAlc of the treatment group had no difference compared with the control group . Compared with the control group, TC and LDL-C of the treatment group was obviously decreased (P < 0.05). And HDL-C was significantly increased ( P < 0 . 05 ) . The carotid artery IMT of the treatment group decreased from ( 0 . 11 ±0 . 01 ) cm to ( 0 . 08 ± 0 . 01 ) cm . And compared with the control group , there was statistical significance ( P <0 . 05 ) . The carotid atherosclerotic plaque area of 58 cases in the treatment group decreased from ( 0 . 37 ±0.56) cm2 to (0.21 ± 0.25) cm2. However, there was no statistical significance compared to the control group. There were 5 adverse events in the control group and 9 adverse events in the treatment group . And there was no difference between two groups. It was concluded that DJC combined with atorvastatin can regulate lipid metabolism and reduce carotid artery IMT .

OBJECTIVETo analyze the relationship between cutaneous glycometabolic disorders and cutaneous neuropathy in diabetic rats, and to look for the mechanism of neuropathy and impaired wound healing.

METHODSEighty male SD rats were randomly divided into the normal control group (NC, n = 20), diabetic group (D, n = 20), aminoguanidine-interfered group (AI, n = 20), and insulin-interfered group (II, n = 20) by drawing lots. Diabetes was reproduced in rats of D, AI, and II groups with intraperitoneal injection of streptozotocin (STZ). Then, rats in AI group were fed with 100 mg×kg(-1)×d(-1) aminoguanidine, while rats in II group were subcutaneously injected with insulin for satisfactory control of blood glucose. Changes in mechanical and heat pain thresholds of pad of hind limb were measured at post injection week (PIW) 2, 4, 8. Skin specimens were collected during PIW 2-8 from pads for determination of contents of glucose, advanced glycation end product (AGE), substance P (SP), calcitonin gene-related peptide (CGRP), and observation of distribution and ultrastructure of skin nerve fibers. Data were processed with t test.

RESULTSThe mechanical and heat pain thresholds in D group at PIW 2 [(6.3 ± 1.5) g, (6.0 ± 0.9) s, respectively ] were obviously lower than those in NC group [(13.0 ± 3.2) g, (10.3 ± 1.2) s, with t value respectively 2.71, 3.42, P values all below 0.05]. Contents of glucose and AGE in skin tissue in D group were significantly increased when compared with those in NC group, especially at PIW 8 [(2.85 ± 0.33) mg/g, (31.7 ± 3.2) U/mg of hydroxyproline vs. (0.82 ± 0.22) mg/g, (22.2 ± 1.9) U/mg of hydroxyproline, with t value respectively 1.65, 6.47, P values all below 0.01]. The myelinated nerve fibers were edematous and degenerated, with axons compressed, while the unmyelinated nerve fibers were vacuolated, with microfilament and microtubule disorderly arranged. Content of SP in skin tissue in D group was lower as compared with that in NC group, especially at PIW 2 [(16.8 ± 3.4) pg/g vs. (28.5 ± 5.0) pg/g, t = 2.42, P < 0.01]. There was no obvious difference in content of CGRP between NC and D groups, and also in content of glucose in skin between D and AI groups. Compared with those in D group, content of AGE in AI group at PIW 8 was decreased markedly [(27.2 ± 1.4) U/mg of hydroxyproline, t = 3.38, P < 0.05]; contents of glucose and AGE in II group at PIW 8 were significantly decreased [(1.42 ± 0.38) mg/g, (23.6 ± 1.3) U/mg of hydroxyproline, with t value respectively 1.74, 8.17, P < 0.05 or P < 0.01]. Compared with that in D group, contents of SP in AI and II groups were increased, with a delay in time of trough value. Content of CGRP showed no obvious difference among D, AI, and II groups.

CONCLUSIONSHigh glucose and accumulation of AGE are key mediators of cutaneous neuropathy and impaired wound healing in diabetes mellitus, which confirms that diabetic wound takes an atypical footing during wound repairing. Aminoguanidine and insulin can reduce contents of glucose and AGE in diabetic skin tissue, and ameliorate diabetic cutaneous neuropathy.

Animals ; Diabetes Mellitus, Experimental ; complications ; metabolism ; Glucose ; metabolism ; Glycation End Products, Advanced ; metabolism ; Male ; Peripheral Nervous System Diseases ; etiology ; Rats ; Rats, Sprague-Dawley ; Skin ; metabolism ; pathology ; Skin Diseases ; etiology ; Wound Healing

AIM:To study the effect of Aurora protein kinase inhibitor VX-680 on homogeneous adhesion and migration ability in human hepatocellular carcinoma cell line HepG 2.METHODS:The HepG2 cell were divided into ex-perimental group and control group, respectively.VX-680 was used in experimental groups at 3 concentrations(3.125 μmol/L group,6.25 μmol/L group and 12.5 μmol/L group).DMSO was used in the control group.The effects of VX-680 at different concentrations on the adhesion ability of human hepatocellular carcinoma HepG 2 cells were observed by cell slow aggregation test and separation experiment.The effects of VX-680 at different concentrations on the migration ability of HepG2 cells was detected by wound healing assay.The expression of E-cadherin in HepG2 cells was detected by Western blot.RESULTS:The results of the slow aggregation test showed that compared with the control group,the number of cell clumps formed in experimental groups was significantly decreased(P<0.01).The results of separation experiment showed that the ratio of NTC/NTEgradually decreased with the increased concentration of VX-680.The results of wound healing as-say showed that as the concentration of VX-680 increased, the cell scratch healing ability gradually weakened compared with control group.The results of Western blot showed that the protein expression of E-cadherin in the HepG2 cells in-creased with the increased concentration of VX-680(P<0.05).CONCLUSION:VX-680 increases the homogeneous ad-hesion and inhibits the migration of HepG 2 cells.

PURPOSE: To compare the diagnostic efficiency of ⁶⁸Gallium labelled prostate-specific membrane antigen positron emission tomography (⁶⁸Ga-PSMA PET) and magnetic resonance imaging (MRI) for staging the lymph node metastases (LNMs) in the prostate cancer.MATERIALS AND METHODS: A broad search of scientific databases including PubMed, EMBASE, Web of Science, Cochrane Database, and Chinese Biomedicine Literature Database (updated prior to November 1st, 2018) was conducted systematically by two reviewers. In this paper, we evaluated the methodological quality of each included article independently and performed a systematic review and meta-analysis to reveal the summary of the diagnostic performance of ⁶⁸Ga-PSMA PET and MRI in properly identifying LNMs of intermediate- and/or high-risk prostate cancer.RESULTS: Thirteen eligible articles comprising 1,597 patients were included. For LNMs detection, the pooled sensitivity and specificity of ⁶⁸Ga-PSMA PET were 0.65 (95% confidence interval [CI]: 0.49–0.79) and 0.94 (95% CI: 0.88–0.97), respectively, while the corresponding values of MRI were 0.41 (95% CI: 0.26–0.57) and 0.92 (95% CI: 0.86–0.95). The area under the symmetric receiver-operating characteristic (SROC) curve for ⁶⁸Ga-PSMA PET and MRI were 0.92 and 0.83, respectively.CONCLUSIONS: In intermediate- or high-risk pre-treatment prostate cancer, ⁶⁸Ga-PSMA PET had a higher sensitivity and a slightly different specificity in probing the LNMs when comparing with MRI. Moreover, the area under the SROC curve indicated that ⁶⁸Ga-PSMA PET was a more effective weapon for predicting the LNMs prior to radical surgery.

OBJECTIVE: To study the effects of Angelicaesinensis radix (Danggui) decoction on the therapeutic action and the colonic morphology and mucus secretion in XuexuBianmi model mice. METHODS: Sixty Kunming mice, maleandfemaleinhalf, were randomly divided into six groups according to gender and weight (=10):normal control group, XuexuBianmi model group, positive control group, and high-dose, middle-dose and low-dose Danggui groups. Except the normal control group, the mice in the rest groups were orally administrated with diphenoxylate (DPN) and subcutaneously injected with acetylphenyhydrazine (APH) and intraperitoneally injected with cyclophosphamide (CPA) to copy XuexuBianmi model. Fromthe 14th day, the mice in Danggui groups were orally administered with different doses of Danggui decoction (16.67、8.33、4.17 g/kg), the mice in positive control group were orally administered with Changtongshu granule (5 g/kg), the mice in XuexuBianmi model group were administered with normal saline (NS) at the same volume (10 ml/kg), once a day for consecutive 28 days. The general status were observed, the first black defecation time (FBDT), the water content in stool and in colon in mice of all groups were tested. And the colon tissue was stained with hematoxylin-eosin (HE) and AB-PAS to observe the changes of colonic morphology and the mucus secretion. RESULTS: Compared with the normal control group, there appeared the XuexuBianmi syndromes in model group as follows, the defecation time (FBDT) was significantly prolonged, the water contentinstool and in colon were decreased (<0.01), the colonic mucosa and gland were atrophied, mucous membrane layer was thinned (<0.01), mucus secretion was decreased. Compared with the XuexuBianmimodel group, the Xuexu and Bianmi syndromes were improved, the FBDT was significantly shortened (<0.05, <0.01), the water content of colon was increased in the three groups of Danggui decoction (<0.05, <0.01). The water content of the stool was obviouslyincreased in groups of 16.67and 8.33kg.dDanggui decoction (<0.05). The atrophy of colon mucosa and gland was improved, the mucus secretion was increased, and the colon lubrication function was improved in the three groups in different doses of Danggui decoction. CONCLUSIONS Dangguican improve the mucosal atrophy, and increase the secretion of colonic mucus, then the stool is softened and promoted to discharge.
Angelica sinensis ; Animals ; Colon ; Drugs, Chinese Herbal ; Mice ; Mucus ; Plant Roots