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 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

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 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

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.

OBJECTIVES: To explore the risk factors for hemorrhagic cystitis (HC) in children with β-thalassemia major (TM) undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT). METHODS: A retrospective analysis was conducted on clinical data of 247 children with TM who underwent allo-HSCT at Shenzhen Children's Hospital from January 2021 to November 2022. The children were divided into an HC group (91 cases) and a non-HC group (156 cases) based on whether HC occurred after operation. Multivariable logistic regression analysis was used to explore the risk factors for HC, and the receiver operating characteristic curve was used to analyze the predictive efficacy of related factors for HC. RESULTS: Among the 247 TM patients who underwent allo-HSCT, the incidence of HC was 36.8% (91/247). Univariate analysis showed age, incompatible blood types between donors and recipients, occurrence of acute graft-versus-host disease (aGVHD), positive urine BK virus deoxyribonucleic acid (BKV-DNA), and ≥2 viral infections were associated with the development of HC after allo-HSCT (P<0.05). Multivariable analysis revealed that incompatible blood types between donors and recipients (OR=3.171, 95%CI: 1.538-6.539), occurrence of aGVHD (OR=2.581, 95%CI: 1.125-5.918), and positive urine BKV-DNA (OR=21.878, 95%CI: 9.633-49.687) were independent risk factors for HC in children with TM who underwent allo-HSCT. The receiver operating characteristic curve analysis showed that positive urine BKV-DNA alone or in combination with two other risk factors (occurrence of aGVHD, incompatible blood types between donors and recipients) had a certain accuracy in predicting the development of HC after allo-HSCT (area under the curve >0.8, P<0.05). CONCLUSIONS Incompatible blood types between donors and recipients, occurrence of aGVHD, and positive urine BKV-DNA are risk factors for HC after allo-HSCT in children with TM. Regular monitoring of urine BKV-DNA has a positive significance for early diagnosis and treatment of HC.
Humans ; Child ; Retrospective Studies ; beta-Thalassemia/therapy* ; Cystitis/epidemiology* ; Hematopoietic Stem Cell Transplantation/adverse effects* ; Risk Factors ; Hemorrhage/etiology* ; Graft vs Host Disease/complications* ; DNA ; Polyomavirus Infections/epidemiology*

Aim of this study was to develop the detection method of soluble human leukocyte antigens I (sHLA-I) and to explore sHLA-I level alteration in storage blood and its significance. sHLA-I level in sera of 60 Guangdong normal individuals and sHLA-I concentration in blood components from 20 donors quantitatively were detected by sandwich ELISA. The results showed that sensitivity of this assay was 2.84 ng/ml. Coefficients of variation were 5.80% within assays and 9.00% between assays respectively. The recovery rate was >/= 98.57%. The sHLA-I level of normal individuals in Guangdong was (699.54 +/- 360.10) ng/ml. sHLA-I in red blood cells stored for 28 days and in random-donor platelets were significantly higher than that in other blood components and their amount was proportionate to the number of residual donor leukocytes and to the length of storage. In conclusion, sandwich ELISA assay for detection of sHLA-I is a sensitive, specific and stable technique. Blood components with different concentration of sHLA-I may be chosen for clinical transfusion.
Apoptosis ; Blood Preservation ; Enzyme-Linked Immunosorbent Assay ; Histocompatibility Antigens Class I ; blood ; Humans ; Sensitivity and Specificity ; T-Lymphocytes, Cytotoxic ; cytology