Objective To investigate the anti-fibrogenesis property and mechanism of short hairpin RNA (shRNA ) targeting heat shock protein 47 (HSP47) in schistosomiasis liver fibrosis mice model . Methods Sixty female BALB/c mice (SPF level) were infected percutaneously with (16 ± 1) Schistosoma japonicum cercariae . Twelve mice which were not infected with Schistosoma japonicum were set as uninfected control .The 60 mice were randomly divided into five groups including 1 mg/kg HSP47-shRNA intervention group ,2 mg/kg HSP47-shRNA intervention group ,4 mg/kg HSP47-shRNA intervention group ,unrelated shRNA intervention group and infected control group ,with 12 mice each group .From 6 to 14 weeks post infection ,mice in HSP47-shRNA intervention groups were intravenously injected with HSP47-shRNA weekly and those in unrelated shRNA intervention group were intravenously injected with 2 mg/kg of unrelated shRNA vector weekly .Survival rate ,liver and spleen in general and liver histology of mice in each group were observed at week 14 which was considered as end of intervention . The expressions of HSP47 mRNA and protein were determined by real-time polymerase chain reaction (PCR) and Western blot .Collagen type Ⅰ ,collagen type Ⅲ ,tissue inhibitor of metalloproteinase-1 (TIMP-1) , matrix metalloproteinase-9 (MMP-9) ,plasminogen activator inhibitor-1 (PAI-1) ,transforming growth factor-β1 (TGF-β1 ) , connective tissue growth factor (CTGF ) , interleukin (IL )-13 and IL-17 at transcriptional level were measured by real-time PCR .Means among groups were compared using student-t test .Results In vivo intervention of HSP47-shRNA mainly localized in hepatic stellate cells of mice . The survival rate of 1 mg/kg ,2 mg/kg and 4 mg/kg HSP47-shRNA intervention groups were 25 .00% , 25 .00% and 33 .33% , respectively .Mice received 4 mg/kg dose of HSP47-shRNA had significantly higher survival rate than the infected controls (χ2 = 4 .168 ,P= 0 .043) .In both 4 mg/kg HSP47-shRNA intervention group and the infected control group , hematoxylin and eosin staining showed chronic granuloma change ,of which ova were wrapped by the spindle-shaped collagen and inflammatory cell infiltrated .The percentage of positive staining for Masson trichrome and sirius red showed the quantity of the collagen deposition was significantly reduced by the HSP 47-shRNA intervention ,compared to the infected control group (t = 3 .191 ,P = 0 .039) .HSP47-shRNA significantly reduced HSP47 expression , and reduced the transcriptional levels of collagen type Ⅰ ,collagen type Ⅲ ,TIMP-1 ,PAI-1 ,TGF-β1 , CTGF ,IL-13 and IL-17 (all P < 0 .01) ,and increased the expression of MMP-9 mRNA ( P < 0 .01) . Conclusion Delivery of shRNA targeting HSP47 to the hepatic schistosomiasis mice model can silence the expression of HSP47 and improve the liver fibrosis .Collagen degradation and related cytokines release through upregulation of MMP-9 and downregulation of PAI-1 may be one of the anti-fibrotic mechanisms in HSP47-shRNA intervention .

Objective: To examine the effects of perioperative intravenous administration of flurbiprofen axetil (FA) on pain associated with transrectal ultrasound-guided prostate biopsy.Methods: This was a randomized,controlled study.Eighty-one patients who underwent 12 core prostate biopsy were included in the study.The patients were randomly assigned to one of three groups (n=27 in each) by type of procedure during prostate biopsy.Group intrarectal local anesthesia (IRLA) received intrarectal 5% (0.05 g/L) lidocaine gel 60 mg, 5 minutes before the procedure alone;Group FA received intravenous flurbiprofen axetil (1 mg/kg) 1 hour before the procedure;Group IRLA+FA received intrarectal 5% lidocaine gel 60 mg, 5 minutes before the procedure and intravenous flurbiprofen axetil (1 mg/kg) 1 hour before the procedure.The patients were asked to score the pain by using visual analogue scale (VAS) in 4 situations,including when the probe was inserted (VASⅠ),during anesthesia (VASⅡ),during biopsy (VASⅢ) and 20 minutes after biopsy (VASⅣ).The findings were evaluated with analysis of variance,and the Tukey post hoc test was followed with an overall 2-tailed significance level at α =0.05.P1, P value between Group IRLA and Group FA;P2, P value between Group FA and Group IRLA +FA,P3, P value between Group IRLA and Group IRLA +FA.The bonferroni method was used to adjust the test level, α=0.017,a P value of less than 0.017 was accepted as the threshold for statistical significance.Results: No major complications,including sepsis and severe rectal bleeding,were noted in any patient.There were no differences in general condition of the patients before procedure among the 3 groups.There were statistically significant differences in VAS scores among the 3 groups in VASⅡ (5.7±2.2, 3.0±1.5,3.3±1.9,respectively,P=0.012) and VASⅢ (6.7±2.3,3.0±2.1,2.9±1.6,respectively,P=0.001).There were no differences in the pain scores among the 3 groups during probe insertion (VASⅠ, 3.2±1.0,4.1±2.1,4.2±1.7, respectively,P=5.752) and 20 minutes after biopsy (VASⅣ, 1.4±2.1,1.0±0.9,1.1±0.7,respectively,P=3.772).Between-column differences among the 3 groups were VASⅡ (P1=0.007,P2=5.655,P3=0.001,respectively) and VASⅢ(P1=0.008,P2=7.517,P3=0.001,respectively),the differences between Group IRLA and Group FA,Group IRLA and Group IRLA +FA in VASⅡ and VASⅢ were statistically significant.Conclusion:The intravenous flurbiprofen axetil was found to be more effective than intrarectal lidocaine gel alone.

Objective: To study the expression of transmembrane protein CMTM2 in the testis and sperm of adult males and to approach the potential function of the protein in the male reproductive system.Methods: The expression of CMTM2 in human testis and sperm was confirmed by Western blot.Immunohistochemical staining was used for detecting CMTM2 localization in the testis tissue, TRITC-CMTM2 and FITC-Hoechst double immunofluorescence staining was performed to examine the subcellular localization of CMTM2 in the human sperm before and after acrosome reaction, that is, immunofluorescent staining was used for detecting CMTM2 localization in both the testis and sperm before and after the acrosome reaction.Results: CMTM2 was presented in both human testis and sperm.In the testis, CMTM2 immunoreactive particles were observed mainly in the membrane of the different stages of spermatogenic cells.In the human sperm, its immunoreactivity was restrictively localized to the posterior head where sperm-egg fusion occurred, and the CMTM2 localization was not affected by sperm acrosome reaction.CMTM2 was widely expressed in seminiferous tubules of the human testis, mainly in the cell membranes of spermatogenic cells, which was consistent with the previous reports.The immunofluorescence performed on frozen human testis slides showed similar findings with immunohistochemistry, which gave weight to the localization of CMTM2 in the cell membranes of spermatogenic cells at different stages.TRITC-CMTM2 and FITC-Hoechst double immunofluorescence staining was performed to examine the subcellular localization of CMTM2 in the human sperm before and after acrosome reaction.CMTM2 was localized at the posterior head of sperm before and after acrosome reaction.The localization and expression of CMTM2 were not affected by sperm acrosome reaction.Conclusion: Expression of CMTM2 in the male reproductive system of the adult human exhibits cell-and region-specific patterns, which suggests that they may play an important role in spermatogenesis and sperm-egg fusion.The expression of CMTM2 in the male reproductive system of the adult human exhibits cell-and region-specific patterns, which suggests that they may play an important role in spermatogenesis and sperm-egg fusion.However, it still remains to be further elucidated about the definite role of CMTM2 in male reproductive system and the process of spermatogenesis.And in vitro fertilization experiments are needed to confirm the role of CMTM2 in fertilization in future.

OBJECTIVETo study the relationship between morphological characteristics of growth cycle for Epimedium acuminatium and accumulation level of secondary metabolites.

METHODAfter making habitat clear, methods of morphology observation, specimen preparation, organism charting were applied to record morphological characteristics of E. acuminatium at different stages. HPLC was employed to detect icariin content of different parts of the plant at the stage of vegetative growth and sexual reproduction.

RESULTThe growth cycle of E. acuminatium was divided into vegetative growth stage and sexual and asexual reproduction coexistence stage. Seven to eight years were needed for the plants flowering and seeding of E. acuminatimcan in open land of forest edge, but within good nutrition conditions, growth cycle is shortened into three to four years. The difference of icariin content in two growth stages was not significant.

CONCLUSIONMorphological change of lower and foliar organ can be utilized to estimate growth age. But after applying sexual propagation for both root and stem of grown plants, it's been difficult to determine their growth year. If individual biomass or yield is economical, plants can be harvested in both vegetative growth and sexual reproduction stages.

Biomass ; Chromatography, High Pressure Liquid ; methods ; Epimedium ; growth & development ; metabolism ; Flavonoids ; metabolism ; Plant Roots ; growth & development ; metabolism ; Reproduction ; Seedlings ; metabolism

OBJECTIVETo observe and compare the protective effect of garlic oil against carbon tetrachloride (CCL)-induced acute liver injury.

METHODSThe experiments include 4 preventive groups and 2 therapeutic groups. In every preventive and therapeutic group, the mice were randomized into 6 groups with 15 each, including one negative control group, one solvent control group, one CCl4 model group and 3 garlic oil groups (25, 50, and 100 mg/kg body weight). Before given a single gavage of CCl4 (80 mg/kg), the mice were pretreated with garlic oil by gavage in preventive group 1 (30 days, once daily), preventive group 2 (5 days, once daily), preventive group 3 (ahead of 2 h, once), preventive group 4 (immediately, once) or the vehicle (corn oil, 10 ml/kg) in solvent control group. In therapeutic groups, the mice were gavaged garlic oil 2 h (once, in therapeutic 1) or for 5 days (once daily, in therapeutic 2) after administration CCl. After 24 h of the last administration, blood was collected and centrifuged at 2500 r/min at 4 degrees C for 10 min, and serum was removed to measure ALT and AST activities. The liver was dissected, weighed to calculate the liver coefficient (relative liver weight). At the same time, the liver samples were studied by histological examinations.

RESULTSCompared with negative group, the liver coefficient and the activities of ALT and AST in serum of model group were increased remarkably (P < 0.01). Compared with CCl model group, the liver coefficient and the activities of ALT and AST in serum were decreased significantly (P < 0.01) by garlic oil dose-dependently in each preventive group. Simultaneously, histological assessment showed that garlic oil effectively alleviated hepatocyte injuries induced by CCl4. Comparing the preventive effects of garlic oil in every group, it was better in preventive group 3 than others. However, all indexes and histological examinations in therapeutic group 1 did not show the difference with those of CCl4 model group. In therapeutic group 2, all indexes recovered after 5 d of CCl4 administration.

CONCLUSIONSGarlic oil can prevent acute liver injury induced by CCl4 and the effect is better in ahead of 2 h group than others.

Alanine Transaminase ; metabolism ; Animals ; Aspartate Aminotransferases ; metabolism ; Carbon Tetrachloride Poisoning ; drug therapy ; prevention & control ; Chemical and Drug Induced Liver Injury ; drug therapy ; prevention & control ; Garlic ; Liver ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; Plant Oils ; administration & dosage ; therapeutic use

OBJECTIVETo explore the effect of anhydrous alcohol sclerotherapy following transrectal ultrasound-guided puncture in the treatment of Mülllerian duct cyst.

METHODSTotally 44 patients with Müllerian duct cyst underwent transrectal ultrasound-guided puncture and sclerotherapy, 21 injected with anhydrous alcohol of half the volume of the aspirated cystic fluid followed by aspiration of all the fluid 5 minutes later (treatment group) , and the other 23 treated by cystic fluid aspiration only (control group). The clinical effects of the two methods were compared.

RESULTSThe response rate and cure rate were 80.95 and 52.38% in the treatment group, as compared with 56.52 and 26.09% in the control (P < 0.001). No adverse events were observed in either of the two groups.

CONCLUSIONTransrectal ultrasound-guided puncture and anhydrous alcohol sclerotherapy is a safe and effective approach to the treatment of Mülllerian duct cyst.

Cysts ; therapy ; Endosonography ; Humans ; Male ; Middle Aged ; Mullerian Ducts ; Rectum ; diagnostic imaging ; Sclerotherapy ; Suction ; methods

Objective Compare the efficacy of superselective arterial embolization with observation in the treatment of HFP caused by trauma.Methods From Jan,2013 to Dec,2017,6 traumatic HFP patients were involved into the study,the age range from 18 to 44 years old,with an average age of 26.2 years.The fistula was measured by doppler ultrasonography before the treatment with its longest diameter,and which ranges from 0.8 to 1.4 cm,the average diameter is 1.14 cm.All patients are divided into two groups according to their treatment.Operation group:4 patients underwent the pudendal artery superselective gelatin sponge vascular embolization.Control group:2 cases were treated with local cold compressing under watchful waiting.In control group,patients are treated by local cold compress with ice bag for 3 weeks:15 mins × 10 times every day.The erectile hardness of the patients was observed and the fistula was evaluated by doppler ultrasonography 3 weeks later,and all the patients were reexamined by doppler ultrasonography 1 year later.The erectile hardness was divided into four grades according to the patient~ subjective feelings:grade Ⅰ,the penis was enlarged but not rigid;grade Ⅱ,the penis had hardness,but not enough to insert into the vagina;grade Ⅲ,the penis could be inserted into the vagina,but not to achieve complete erection;grade Ⅳ,the penis fully erect and firm.The erectile function was evaluated with the International Index of Erectile Function-5(IIEF-5) after 1-year's follow-up.Results 4 patients in the operation group were operated successfully without obvious complications after operation.Median follow-up time of all the patients was 21 months (5-50).In group operation,4 cases of surgical patients did not have the recurrence of priapism.There was no obvious abnormality in the color doppler ultrasound of penis.The score of IIEF-5 was 25,24,24 and 23,respectively.Postoperative erectile function made a complete recovery.In group observation,2 patients under watchful waiting still have grade Ⅱ-Ⅲ sustained priapism,IIEF-5 score is 21 and 19 respectively,the penis color doppler ultrasound scan still showed the change of arteriovenous fistula.Conclusion Superselective arterial embolization is effective in the treatment of HFP,and the recovery of sexual function is better after operation compared with watchful waiting.

Objective To study the effects of Jianpi Xiaoai Prescription on cell cycle and apoptosis of human colon cancer HCT116 cells and related factors of Wnt/β-catenin signaling pathway; To investigate its mechanisms of anti-metastasis and recurrence of colorectal cancer. Methods The logarithmic growth phase HCT116 cells were divided into blank group, saline group, 5-Fu group, and Jianpi Xiaoai Prescription low-, medium-, and high-dose groups. After intervention for 48 h, the cells were harvested, and the cell cycle and apoptosis were detected by flow cytometry. The protein expression of β-catenin in the nucleus was detected by Western blot, and the expression of c-myc and cyclinD1 mRNA was detected by PCR. Results Compared with the blank group and saline group, the ratio of HCT116 cells apoptosis of Jianpi Xiaoai Prescription low-, medium-, and high-dose groups increased; the proportion of cells in phase G1 increased and the proportion of S cells decreased; the expression of β-catenin protein in the nucleus and the expression of c-myc,cyclinD1 mRNA decreased,especially in the Jianpi Xiaoai Prescription high-dose group,with statistical significance(P<0.05,P<0.01).Conclusion Jianpi Xiaoai Prescription can promote apoptosis of human colon cancer HCT116 cells and block the cell cycle, and its mechanism may be related to regulating Wnt/β-catenin signaling pathway.

OBJECTIVETo investigate the effects of tacrolimus (FK506) on behavioral function and heat-shock proteins (HSP70) expression in nervous tissues of acrylamide (ACR)-induced rats.

METHODSTotally 40 health Wistar rats were randomly divided into control group, model group, low and high doses of FK506 groups. All four groups were treated five times per week for four weeks. Gait score was measured every week. And rats were sacrificed on day 28, the cerebrum, spinal cord and sciatic nerve were dissected, and homogenized in ice bath, then the levels of HSP70 and Bcl-2, Bax were analyzed by western bloting.

RESULTSCompared with the ACR model group, the gait score in low and high doses of FK506 groups decreased by 30.1% and 47.7% respectively in the 4th week. In the cerebrum and sciatic nerve pellet, the level of HSP70 in the FK506 groups increased by 11.6%, 33.3% and 56.3%, 58.5% (P < 0.01), but no significant changes existed in spinal cord. The level of Bcl-2 in the sciatic nerve pellet increased by 39.1% (P < 0.01) but no significant changes existed in the cerebrum and spinal cord from low dose of FK506 group. And the level of Bax in the spinal cord pellet markedly increased by 46.8% but not in cerebrum and sciatic nerve pellet; Whereas in the tissues mentioned above, the levels of Bcl-2 were enhanced remarkably by 16.3%, 14.8% and 56.0% (P < 0.01) in the high dose of FK506 group. And the level of Bax in the cerebrum and spinal cord pellet markedly increased by 16.4% and 40.2% but not in sciatic nerve. The values of Bcl-2/Bax in low and high doses of FK506 groups clearly increased by 15.9%, 33.3%, 36.9% and 30.1%, 49.1%, 60.1% (P < 0.01).

CONCLUSIONThe administration of FK506 has dramatically neuroprotective effects against the development of ACR neuropathy, which may be related to up-regulating the expression of HSP70 and Bcl-2 with down-regulating the expression of Bax.

Acrylamide ; poisoning ; Animals ; HSP70 Heat-Shock Proteins ; metabolism ; Male ; Nerve Tissue ; drug effects ; metabolism ; Neuroprotective Agents ; therapeutic use ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Wistar ; Tacrolimus ; therapeutic use ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo investigate the dynamic changes of neurofilaments (NFs) proteins in spinal cords of hens with phenylmethylsulfonyl fluoride (PMSF) pretreatment for exploring the mechanism of tri-o-cresyl phosphate (TOCP)-induced delayed neuropathy (OPIDN).

METHODAdult Roman hens were randomly divided into three groups, control, TOCP and PMSF + TOCP. Birds in PMSF + TOCP set were pretreated with PMSF, 24 hours later, hens in both TOCP group and PMSF + TOCP group were administrated with TOCP at a single dosage of 750 mg/kg. Then all animals were sacrificed on the corresponding time-points of 1, 5, 10, and 21 days respectively after dosing of 750 mg/kg TOCP. The spinal cords were dissected, homogenized, and centrifuged at 100,000 x g. The levels of high molecular neurofilament (NF-H), medium molecular neurofilament (NF-M) and low molecular neurofilament (NF-L) in both pellet and supernatant fractions of spinal cords were determined by SDS-PAGE and Western-blotting.

RESULTSThe hens in TOCP group showed paralysis gait at the end of 21-day experimental period. The levels of NFs proteins in spinal cords changed obviously. Compared with control, the NFs in pellet showed a dramatic decrease on day 10 and then followed by a recovery. In the supernatant, the NFs proteins showed similar changes, which decreased significantly on day 10 and almost recovered control on day 21. Such as, NF-L, NF-M and NF-H decreased by 51%, 86% and 38% on day 10. The OPIDN signs were not observed in PMSF + TOCP group, and imbalances of NFs were obviously alleviated. Compared with control, only NF-M in pellet increased by 21% (P < 0.05) on day 21, others remained no changes; The levels of NF-H and NF-M in supernatant respectively increased by 19% and 35% on day 21, others were no significant statistical differences.

CONCLUSIONTOCP may induce imbalance of NFs levels in progress of OPIDN, and PMSF pretreatment may protect animals from OPIDN by reducing above changes, which may explain that TOCP-induced imbalance of NFs may be connected with the occurrence and development of OPIDN.

Animals ; Chickens ; Female ; Neurofilament Proteins ; drug effects ; Phenylmethylsulfonyl Fluoride ; pharmacology ; Protein Subunits ; drug effects ; Spinal Cord ; drug effects ; metabolism ; pathology ; Tritolyl Phosphates ; toxicity