1.Inhibiting effects of three components of Astragalus membranaceus on oxidative stress in Chang Liver cells.
Jian LI ; Lin HAN ; Yu-fang MA ; Yi-fan HUANG
China Journal of Chinese Materia Medica 2015;40(2):318-323
The main objective of this research is to investigate the effects of astragaloside IV, calycosin separately glucoside, formononetin on oxidative stress in Chang Liver cells induced by H2O2. In the experiments, Chang Liver cells (a kind of normal human hepatocytes) were used as the research object, bifendate which has a clear hepatoprotective effect was used as the positive control drug, then the oxidative damage model of Chang Liver cells were established by H2O2. Cells were divided into six groups: blank control group, oxidative stress group, astragaloside IV group, calycosin separately glucoside group, formononetin group and positive control group. Then endogenous antioxidant system related indexes were detected by micro plate and colorimetric method; intracellular reactive oxygen species (ROS) were detected by DCFH-DA fluorescent probe; and the expressions of CYP2E1 were evaluated by liver microsomes, mRNA, and protein, respectively with spectrophotometry, Real-time PCR method, and Western blot technique. Results showed that H2O2 decreased antioxidant activity, and increased ROS level and expression of CYP2E1. The above oxidative stress status had been changed with protections of the three components of Astragalus membranaceus (compared with oxidative stress group, P < 0.05, P < 0.01), which taken as a whole had equivalent effects as the drug of positive control group( bifendate). Taken together, three Astragalus membranaceus ingredients all had significant or extremely significant inhibiting effects on oxidative damaged Chang Liver cells which were induced by H2O2, and the oxidative damage of Chang Liver cells had been relieved.
Astragalus membranaceus
;
chemistry
;
Cells, Cultured
;
Cytochrome P-450 CYP2E1
;
metabolism
;
Humans
;
Isoflavones
;
pharmacology
;
Liver
;
drug effects
;
Oxidative Stress
;
drug effects
;
Reactive Oxygen Species
;
metabolism
;
Saponins
;
pharmacology
;
Triterpenes
;
pharmacology
2.Effect of prostate volume on histopathological outcomes in patients after laparoscopic radical prostatectomy
Fan ZHANG ; Yi HUANG ; Min LU ; Lulin MA ; Guoliang WANG
Chinese Journal of Urology 2012;33(5):360-363
ObjectiveTo compare the clinical characteristic and postoperative histopathological parameters in different prostate size and to assess the effect of prostate volume on histopathological outcomes in patients after laparoscopic radical prostatectomy. MethodsTwo hundred and sixteen patients from 2006 to 2011 who were proved prostatic carcinoma by biopsy and performed laparoscopic radical prostatectomy wewe retrospectively evaluated.All patients were divided into the small group ( < 30 ml,103 cases),intermediate group(30 -60 ml,71 cases) and large group ( >60 ml,42 cases) according to prostate volume.Clinical variables included age,body mass index (BMI),preoperative PSA level,prostate volume,percentage of positive needles,clinical stage,and biopsy Gleason score (GS).Histopathological parameters included pathological GS,upgrading or not compared to biopsy GS,pathological stage and surgical margins status.Preoperative clinical variables and postoperative histopatholgogical parameters were compared among the 3 prostate groups. ResultsPatients with smaller prostates had lower PSA levels than those with intermediate and larger prostates (P =0.000).They also had worse histopathological outcomes such as pathological GS ( P =0.034 ),upgrading of GS ( P =0.037 ),and pathological stage ( P =0.025 ).35.0% of patients in smaller prostate group had a positive surgical margin compare to 33.8% and 19.0% in intermediate and larger prostate groups,but there was no significant difference (P =0.152).ConclusionMen with smaller prostate may have more high-grade cancer and more advanced disease after laparoscopic radical prostatectomy.
3.Effect of MG132 on the expression of ERK1/2 and connective tissue growth factor in rat peritoneal mesothelial cells induced by high glucose
Wenyan DENG ; Jianfei MA ; Yi FAN ; Lixia SUN ; Lina YANG
Chinese Journal of Nephrology 2013;(3):195-198
Objective To observe the effect of MG132 on the expression of extracellular regulated kinase 1/2 (ERK1/2) and connective tissue growth factor (CTGF) in rat peritoneal mesothelial cells (RPMCs) induced by high glucose.Methods RPMCs were isolated,cultured and passaged by trypsin,then identified.The second generation of cultured RPMCs were used in the experiment.RPMCs were divided into normal control group,high glucose (1.5%,2.5%,4.25%) for 24 hours,high glucose (2.5%) for 0,12,24,48 hours,incubated with MG132 (0.5,1,2 μmol/L) for half an hour and then with high glucose (2.5%) for 24 hours.ERK1/2 protein was detected by Western blotting,and CTGF protein in supernatant was detected by ELISA.Results Compared with the control group,the expression of p-ERK1/2 was significantly increased in the groups stimulated by high glucose (P <0.01),reached the peak at 24th hour (P < 0.01),and then the expression decreased at 48th hour,but still was higher than that in the normal control group (P < 0.01).CTGF protein expression of RPMCs induced by high glucose increased,in time-and dose-dependent manner (P < 0.05).MG132 could significantly decrease the expression of ERK1/2 and CTGF induced by high glucose (P<0.05).Conclusions MG132 can decrease the expression of p-ERK1/2 and CTGF in RPMCs induced by high glucose.The ubiquitin proteasome pathway participates in the development of peritoneal fibrosis,and blocking the way may contribute to the prevention of peritoneal fibrosis.
4.Clinical analysis of permanent 125I prostate brachytherapy monotherapy for early prostate cancer ( report of 18 cases)
Zhicao ZHANG ; Yi HUANG ; Lulin MA ; Fan ZHANG
Chinese Journal of Urology 2012;33(1):55-57
Objective To evaluate the clinical efficacy of permanent 125I prostate brachytherapy monotherapy in patients with early prostate cancer. Methods A total of 18 patients with prostate cancer was diagnosed by prostate biopsy,with Gleason score 5 - 6,average PSA 5.09 ng/ml (range,0.38 -8.73 ),and clinical stage T1c - T2a.All patients underwent 3-dimensional conformal brachytherapy with transperineal ultrasound-guide 125I seeds prostate implantation. Results None of the patients had any adverse events during the operation processes.The operation time was from 37 to 52 minutes and postoperative hospitalization time was from 3 to 4 days.The mean number of implanted 125I seeds was 54.All patients were followed up for 3 to 57 months with a mean of 15 months,while one patient was lost during followingup.The survival rate of PSA progression free was 100% (17/17).The PSA was under 0.2 ng/ml for a long time in the 7 cases who had been followed-up for more than 18 months.The PSA did not show any rising trend in all 17 cases.No severe complications occurred in all cases. Conclusions Brachytherapy monotherapy with 125I seeds implantation is safe and effective for patients with early prostate cancer.
5.Impact of learning curve on perioperative complications in laparoscopic radical prostatectomy: a single center experience of 200 consecutive cases
Lulin MA ; Fan ZHANG ; Yi HUANG ; Guoliang WANG
Chinese Journal of Urology 2015;36(8):611-614
Objective To assess the impact of learning curve on perioperative complications in laparoscopic radical prostatectomy (LRP) by a series of consecutive 200 cases in a single center.Methods Two hundred patients who were biopsy proved prostatic carcinoma and underwent LRP were retrospectively evaluated.All the operations were performed by professor Ma Lulin.The patients were divided into 4 groups according the time of operation.All the clinical data,including age,body mass index (BMI),preoperative PSA level,biopsy Gleason score,and clinical stage were collected.Operative parameters and peri-operative complications were evaluated,including operative time,blood loss,transfusion rate,and perioperative complications.The complications were evaluated by Clavien classification system.Results Age,BMI,PSA,prostate volume,biopsy Gleason score and clinical stage had no significant difference among the 4 groups (P > 0.05).The operative time was gradually lowered from group one to group four.Significantly less blood loss occurred after every 50 cases of LRP (P < 0.001).The complication rates were 15.5% in this series,28.0%,18.0%,12.0% and 4.0% for group one to group four respectively.The complication rate reduced as experience was gained (P =0.008).The rates of mild complications were 24.0%,14.0%,6.0% and 4.0%,and severe complication rates were 4.0%,4.0%,6.0% and 0,respectively.Conclusion With experience was gained and technique was improved,the operative time and blood loss of LRP decreased and complication rate reduced substantially.
6.Observation on postoperative rehabilitation of patients with polypoid lesions of gallbladder treated by ;laparoscopy combined with choledochoscopy
Canye MA ; Baojin LI ; Jiansong LIAO ; Shaofeng FAN ; Zhiyong YI
The Journal of Practical Medicine 2016;32(12):1979-1982
Objective To observe the effects of laparoscopy combined with choledochoscopy in the treatment of polypoid lesions of gallbladder and the effects on postoperative rehabilitation of patients. Methods 78 cases of patients with polypoid lesions of gallbladder treated in our hospital from August 2011 to November 2014 were selected as the study object. According to the order of medical treatment and hospital admission , the patients were numbered and randomly divided into laparoscopy group and combined group , with 39 cases in each group. The laparoscopy group were treated by laparoscopic cholecystectomy while the combined group were treated by laparoscopy combined with choledochoscopy gallbladder-protection surgery. The operation indicators , recovery of gastrointestinal function and incidence of postoperative complications in the two groups were observed. After discharge , the 2 groups of patients were followed up for 1 year. The improvement of postoperative digestive tract symptoms and rehabilitation of patients was observed. Results The intraoperative bleeding volume, postoperative fasting time, defecation time and hospitalization time of the combined group [(20.13 ± 7.48) mL, (0.51 ± 0.98) d, (1.20 ± 0.84) d, (1.98 ± 1.54)d] were less than those of the laparoscopy group (P <0.05); The incidence rate of complications in the combined group (10.26%) was lower than that in the laparoscopy group (33.33%) (P < 0.05); Immediately at the end of operation and 1 week after operation, VAS scores of the combined group [(4.23 ± 0.41), (2.46 ± 0.17) ] were lower than those in the laparoscopy group (P < 0.05); At the last follow-up, KPS score of the combined group increased to (89.98 ± 1.24), which was higher than that in the laparoscopy group (P < 0.05); GradeⅠpostoperative recovery in the combined group accounted for 87.18% which was higher than that in the laparoscopy group (64.10%). There was no case of grade III and the proportion was lower than that in the laparoscopy group (15.38%). The differences in proportions of grade I and III between the two groups were statistically significant (P < 0.05). Conclusion To adopt laparoscopy combined with choledochoscopy in the treatment of polypoid lesions of gallbladder , the curative effect is certain and the minimally invasive advantage is obvious. The long-term recovery effect is good and the safety is high.
7.Simultaneous Determination of Five Active Components in Pifukang Lotion by HPLC with Multiple UV Wavelengths
Na MA ; Guiqiang FAN ; Yi CAO ; Shanhou QI
China Pharmacist 2017;20(5):963-965
Objective: To establish an HPLC method for the simultaneous determination of five active components (paeoniflorin, liquiritin, ammonium glycyrrhizinate, edomin and osthole)in Pifukang lotion with multiple UV wavelengths.Methods: A Diamonsil C18 (200 mm×4.6 mm,5 μm) column was used with the mobile phase consisting of 0.1% hydrochloric acid and acetonitrile with gradient elution.The flow rate was 1.5 ml·min-1, and the column temperature was 30℃.Paeoniflorin was detected at 232 nm(0-6 min), liquiritin was detected at 277 nm(6-10 min),and ammonium glycyrrhizinate, edomin and osthole were detected at 254 nm(after 10 min).Results: The linear range of paeoniflorin,liquiritin,ammonium glycyrrhizinate, edomin and osthole was 28.200-282.000 μg·mL-1(r=0.999 7),12.150-121.500 μg·ml-1(r=0.998 8),13.420-134.200 μg·ml-1(r=0.999 5),0.047-0.466 μg·ml-1(r=0.999 9) and 2.38-23.8 μg·ml-1(r=0.999 9), respectively.The average recovery (RSD) was 98.49%(0.71%), 99.00%(0.62%), 98.38%(0.85%), 97.36%(0.92%) and 97.70%(0.78%), respectively (n=6).Conclusion: The established method is simple, accurate, highly sensitive and well reproducible, which can be used for the determination of the five active ingredients in Pifukang lotion.
8.Combined brachytherapy with intermittent hormonal therapy in treating clinical moderate and high risk non-metastatic prostate cancer
Fan ZHANG ; Yi HUANG ; Lulin MA ; Junjie WANG ; Weiqiang RAN
Chinese Journal of Urology 2017;38(6):448-452
Objective To investigate the clinical value of 125I particle implantation brachytherapy combined with intermittent hormonal therapy for treating clinical moderate and high risk non-metastatic prostate cancer.Methods A prospective study was proceeded and 100 cases with moderate and high risk (cT≥T2b,Gleason score ≥ 7,pre-biopsy PSA ≥ 10 ng/ml)non-metastatic prostate cancer were included.The selected patients were divided into two group.In the study group,patients were treated with 125I particle implantation combined with intermittent hormonal therapy.In the control group,patients were treated with only intermittent hormonal therapy.Hormonal therapy was maximal androgen blockage for two groups,including bicalutamide 50 mg oral every day and Leuprorelin 3.75 mg subcutaneous injection every 28 days.There were 50 cases in each group and clinical trial agreements were signed.During follow-up,PSA were tested every month.Chest X-ray and whole-body hone scanning were checked every 6 months.Hormonal therapy was stopped when patient's PSA level fell to 0.2 ng/ml,and keep stabilized for 3 months.When PSA level elevated for 3 times continuously and over 1 ng/ml,hormonal therapy was initiated again.The IPSS scores were documented before treatment and every 3 months after treatment.Adverse reactions of urinary tract and rectum were assessed every 3 months after 125I particle implantation in study group.The ratio of the first time to stop hormonal therapy,the time duration of first hormonal therapy and stable phase,re-hormonal therapy free survival rate,bone metastasis free survival rate,castration resistance prostate cancer(CRPC) free survival rate,cancer-specific free survival rate and overall survival rate were compared.Results The 100 cases in this study were followed up for 24-40 months,with an average time of 31.6 months.In study group,the PSA level in all cases descended to the level of stopping hormonal therapy.The time duration of hormone therapy ranged from 4 to 12 months,with an average time of 6.3 months.21 (42%) cases had a PSA elevation again to restart hormonal therapy.In control group,the PSA level in 47 cases descended to the level of stopping hormonal therapy.The time duration of hormone therapy ranged from 5 to 15 months,with an average time of 7.2 months.34 (68%) cases had a PSA elevation again to restart hormonal therapy.There was no significant difference in percentage of cases of stopping hormone therapy and in time duration of hormonal therapy for the first cycle.Instead,there were significant differences in stable phase after first cycle hormonal therapy between two groups (27.2 months vs.17.7 months;P < 0.001).When analyzed by Kaplan-Meier survival curve,there was no significant difference in cancer-specific survival rate and overall survival rate.There were significant differences in Re-hormonal therapy free survival (P =0.002),bone metastasis free survival (P =0.04) and CRPC free survival(P =0.005).Conclusions Compared with intermittent hormonal therapy alone,125I particle implantation brachytherapy combined with intermittent hormonal therapy could prolong the hormonal sensitive time in moderate and high risk nonmetastatic prostate cancer patients and control the progress of the prostate cancer.
9.Impact of Xuebijing injection on the expression of tumor necrosis factor α and high mobility group box 1 in rat peritoneal mesotheliai cells induced by lipopolysaccharide
Shuo CHEN ; Yi FAN ; Jianfei MA ; Lina YANG ; Xiuli ZHANG
Chinese Journal of Nephrology 2010;26(1):34-38
Objective To observe the effect of Xuebijing injection on the expression of tumor necrosis factor-alpha (TNF-α) and high mobility group box-1 protein (HMGB-1) in rat peritoneal mesothelial cells (PMCs) induced by lipopolysaccharide (LPS). Methods PMCs were isolated from rat colic omentum and the 3rd generation cells were used in the experiment. PMCs were incubated with LPS at different concentrations (1,10,100 mg/L);with LPS (10 mg/L) for 2, 6, 12, 18, 21, 24, 36 h;with Xuebijing injection at different concentrations (2,10,20 g/L) after incubation with LPS (10 mg/L) for 2 h. PMCs in the control group were incubated with medium. HMGB-1 mRNA was detected by RT-PCR. TNF-α and HMGB-1 protein in supernatants was detected by ELISA. Results Compared to the control group, the expression of HMGB-1 mRNA and protein was significantly increased in groups stimulated by LPS in a time- and dose-dependent manner (all P<0.05);the expression of TNF-α was increased in the groups stimulated by LPS in a dose-dependent manner (P<0.05). In the groups stimulated by LPS (10 mg/L), the expression of TNF-α appeared double hump within 36 hours. Compared to LPS (10 mg/L) group, Xuebijing injection significantly inhibited the expression of HMGB-1 and TNF-α (all P<0.05 ) in a dose-dependent manner. Conclusions HMGB-1 as a late mediator of inflammatory responses may play a role in the pathogenesis of peritoneal dialysis related peritonitis. Xuebijing injection can reduce peritoneal inflammatory impairment by inhibiting the up-regulation of TNF-α and HMGB-1 induced by LPS.
10.Construction of eukaryotic vector of monkey B virus glycoprotein D gene and the gD gene expression
Xin WANG ; Simeng YI ; Huifang LIU ; Kai MA ; Junwen FAN ; Yunan MA ; Ying YOU ; Zhaozeng SUN
Chinese Journal of Comparative Medicine 2015;(6):28-31,81
Objective To establish an eukaryotic vector of monkey B virus glycoprotein D gene and analyze the expression of gD gene in human embryonic kidney 293T cells.Method First, the protein of monkey B virus glycoprotein D was obtained by gene synthesis.The gene fragments were digested with Pst I and Not I, and ligated to pEGPF-N3. Then, the recombinant plasmid pEGPF-N3-GD was transfected into 293T cells.The expression of gD protein in the cells was detected by Western blot, and the expression localization was investigated using laser scanning confocal microscopy. Results The recombinant plasmid pEGPF-N3 carrying gD gene was successfully constructed, and normally expressed in the 293T cells.Conclusions Glycoprotein D of monkey B virus is expressed successfully in the 293T cells and the protein is located on the cell surface.It may be useful for the preparation of specific recombinant antigen to the glycoprotein D of monkey B virus on cell surface, and can be also used for preparation of antigen slide for detection of monkey B virus.