1.Small Interfering RNA of Cystathionineβ-Synthase/Cystathionineγ-Lyase-Derived Hydrogen Sulfide Synthesis Induces Apoptosis of Rat Hepatic BRL Cell Line
Jun YAN ; Yanmin LI ; Xuexi GUO ; Mingfang ZHANG
Tianjin Medical Journal 2014;(9):853-858
Objective To explore the inhibitory effects of endogenous hydrogen sulfide, a novel and important gas-eous transmitter generated in mammalian tissues mainly by cystathionine β-synthase (CBS) or cystathionineγ-lyase (CSE) on the apoptosis of the rat hepatic BRL cell line in physiological condition. Methods BRL cells were cultured, and divid-ed randomly into several groups in different phases of the experiment, including negative-siRNA (control) group, CBS siRNA (CBS 1~3) group and CSE siRNA (CSE 1~3) group, which were used to select the most efficient sequences of siRNAs at 48 or 24-hour transfection. Solution group and (CBS+CSE) siRNA group were added to detect the variation of apoptosis. The BRL cell line was observed and evaluated at 0, 4, 8, 12, and 24 hrs after siRNA transfection. When the mechanisms of the apoptosis were detected, CBS/CSE siRNAs were transfected individually or jointly into BRL cells, and compared with nega-tive-siRNA group to examine the variation. The genic and protein expression of CBS/CSE were detected by RT-PCR and Western blot assay. After transfection of CBS/CSE siRNA, the apoptosis of BRL cells was detected by Hoechst stain and flow cytometry (FCM). The mitochondrial membrane potential (MMP) changes were observed by fluorescent staining. Western blot assay was used to examine the protein expression of intracytoplasm cytochrome C (Cyt C) and cleaved-caspase 3. Re-sults CBS and CSE were observed in BRL cells. After transfection of CBS/CSE siRNA, endogenous H2S generation de-creased and the apoptosis of BRL cells increased. Accordingly, the expression of intracytoplasm-Cyt C and cleaved-caspase 3 increased. Conclusion The inhibition of endogenous H2S synthesis induced the apoptosis of BRL cells under physiologi-cal condition, which may be involved in mitochondrial pathway of apoptosis.
2.Effects of fuzheng yiliu granules on apoptotic rate and mitochondrial membrane potential of hepatocellular carcinoma cell line H22 from mice
Jianxiong ZHAO ; Fulian GUO ; Decheng BAI ; Xuexi WANG
Journal of Integrative Medicine 2006;4(3):271-4
OBJECTIVE: To evaluate the effects of Fuzheng Yiliu Granules (FZYLG) on apoptotic rate and mitochondrial membrane potential (Delta psi m) of hepatocellular carcinoma cell line H22 from mice. METHODS: Forty-eight mice inoculated with H22 cells were randomly divided into four groups: untreated group, cyclophosphamide-treated group, high-dose FZYLG-treated group and low-dose FZYLG-treated group. After 14 days of corresponding treatment, H22 cells in each group were stained with propidium iodide, and the apoptotic rates were detected by flow cytometry (FCM). The rhodamine 123 was used as a fluorescence probe to label the H22 cells, and the fluorescence intensities were observed with laser scanning confocal microscope. The fluorescence intensity of H22 cells indicated the Delta psi m of H22 cells. RESULTS: FZYLG could significantly increase the apoptotic rate while reduce the Delta psi m of H22 cells from mice as compared with those in the untreated group. CONCLUSION: The antitumor effects of FZYLR on H22 cells from mice are related to decreasing the Delta psi m and then inducing the apoptosis of the H22 cells.
3.Risk factors of BK virus infection post renal transplantation
Yingxin FU ; Wenli SONG ; Chunbai MO ; Gang FENG ; Xuexi GUO ; Haiming ZHANG ; Zhenglu WANG ; Weiping ZHENG ; Hong ZHENG ; Zhongyang SHEN
Chinese Journal of Urology 2009;30(12):809-812
Objective To analyze the risk factors of BKV infection and compare the real-time PCR procedure and urinary sediment smears of patients checked for decoy cells. Methods The peripheral blood samples of 129 renal recipients were collected. According to the result of PCR, 129 patients were divided into 2 groups:①BKV-DNA(+);②BKV-DNA(-). The sex, age, cold ischemia time, hemotodialysis duration, immunosuppressive agent and other clinical parameters were compared between the 2 groups and a Logistic regression was performed to analyze the risk factors of BKV infection. Results There were 20(15. 5%) patients in BKV-DNA(+), 109(84. 5%)patients in BKV-DNA(-)group. Logistic regression found that the cold ischemia time, hematodialysis duration, living donor were significantly related to the BKV-DNA. The results of the real-time PCR procedure and urinary sediment smears of patients checked for decoy cells were related. Conclusion Real-time fluorescent quantitative PCR and urine decoy cell are good way for detection of BKV infection after renal transplantation. The cold ischemia time and hematodialysis duration and brain death donor were the risk factors of BKV infection post renal transplantation.
5.Clinicopathological features analysis of common complications in pediatric liver transplantation
Zhenglu WANG ; Zhandong HU ; Wenjuan CAI ; Chao SUN ; Wei GAO ; Zhiqi YIN ; Xuexi GUO ; Yan LI ; Mingfang ZHANG ; Lei LIU ; Zhongyang SHEN
Chinese Journal of Organ Transplantation 2018;39(9):527-533
Objective The purpose of this study was to explore common complications and their clinicopathological features in pediatric liver transplantation.Methods Clinical and pathological data of 240 liver biopsies from 168 children that conducted liver puncture from January 2015 to May 2018 in Tianjin First Central Hospital was retrospectively analyzed.We comprehensively analyzed incidence rate and pathological features of various complications,and correlations between acute rejection and C4d staining result or Banff score.Results A total of 86.67% (208/240) liver biopsies could be definitely diagnosed with incidence rate of main complications in descending order as follows:T cell mediated rejection (TCMR) 60.57% (126/208),drug-induced liver injury (DILI) 17.31% (36/208),biliary complication 8.17% (17/208),vascular complication 3.37% (7/208),ischemia/reperfusion injury (IRI) 2.88% (6/208),antibody mediated acute rejection (AMR) 1.92% (4/208),HBV infection 1.92% (4/208),non-alcoholic fatty liver disease (NAFLD) 1.44% (3/208),chronic rejection (CR) 0.96 % (2/208) and HCV infection 0.48 % (1/208).TCMR and AMR in acute rejection (AR) accounted for 96.92% (126/130) and 3.08% (4/160),and into(portal-based,PB)type TCMR accounted for 96.03%(121/126) with the detectable rate of BP type subtype TCMR of 26.45%(32/121)within 30 d.There were 65.87% (83/126)、25.40% (32/126) 和4.76% (6/126) of BP TCMR samples with "Banff ACR RAI" score within 3-5,6-7 and 8-9,and RAI score was negatively correlated with postoperative time (r =0.127,P =0.084).The incidence rate of central perivenulitis (CP) and portal eosinophils infiltration (PEI) in BP TCMR was 63.63% (77/121) 和43.80% (53/ 121),respectively,additionally,the PEI level was positively correlate with RAI score (P<0.05).CP TCMR and AMR occurred within 30d-365 d and 8 d-180 d,respectively postoperative,while,the two CR occurred at 1095 d and 1335 d postoperative,and significant correlation was strikingly observed between rejection subtype and postoperative time (Z =9.231,P =0.026).C4d positive rate was 10% (24/240),which was associated with Banff score and postoperative time,besides,C4d score was also correlated with rejection subtype and RAI score.The occurrence of DILI was mainly at time of <90 d or >180 d postoperative,and the detectable rate of biliary complication within 180 d postoperative was 82.35% (14/17),IRI Appear in <30d.Hepatic artery complication account for nearly 57.14% (4/7),occurrence time is ≤90 d.Occurrence of HBV infection,CMV infection and NAFLD were mainly at >365 d,<90 d and <365 d,respectively.Conclusion There were lots of differences in clinical and pathological features among multi pediatric liver transplantation complications.Liver puncture plays an important role in rejection subtype classification and grading,as well as in non-rejection complications identification.
6.Clinical characteristics and prognostic analyses of cervical neuroblastoma
Qiaoyin LIU ; Shengcai WANG ; Yaqiong JIN ; Ping CHU ; Yongli GUO ; Xiaoli MA ; Yan SU ; Jie ZHANG ; Yanzhen LI ; Xuexi ZHANG ; Nian SUN ; Zhiyong LIU ; Xin NI
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2022;57(6):711-717
Objective:To determine the characteristics of cervical neuroblastoma and the effect of resection extent on survival and outcomes.Methods:We performed a retrospective review of 32 children with cervical neuroblastoma treated at Beijing Children′s Hospital between April 2013 and August 2020. Data were collected from the medical record. The individualized therapy was designed based on staging and risk group. Based on the extent of resection, patients were divided into incomplete and complete resection groups. Event free and overall survival rates were compared between two groups using the Kaplan-Meier method.Results:The ages of patients ranged from 1 month to 81 months, with a median age of 11 months, including 7 males and 15 females. Twenty-nine patients (90.6%) presented with cervical painless mass. The average diameter of the primary tumors was (5.12±1.43) cm. Tumors were located in the parapharyngeal space in 25 cases (78.1%) and in the root of the neck in 7 cases (21.9%). None had MYCN amplification. According to International Neuroblastoma Staging System (INSS), 15 patients (46.9%) were identified as stage 1, 11 patients (34.3%) as stage 2B, 3 patients (9.4%) as stage 3 and 3 patients (9.4%) as stage 4. There were 12 patients (37.5%) at low risk, 17 patients (53.1%) at intermediate risk and 3 patients at high risk according to Children′s Oncology Group (COG) risk classification system. All patients underwent tumor resection. Postoperatively Horner′s syndrome occurred in 13 patients (40.6%), pneumonia in 9 patients (28.1%), pharyngeal dysfunction in 8 patients (25.0%) and transient hoarseness in 4 patients (12.5%). At a median follow-up of 36.5 months, the overall survival rate was 96.4%, with no significant difference between incomplete and complete resection groups (100.0% vs. 96.3%, χ 2=0.19, P=0.667); the event free survival rate was 78.1%, with a significant difference between the two groups (40.0% vs. 85.2%, χ2=6.71, P=0.010). Conclusions:Primary cervical neuroblastoma has a young onset age, mostly in low and medium risk groups, and represents favorable lesions with good outcomes after multidisciplinary therapy. Less aggressive surgery with preservation of important structures is recommended. Complete resection should not be attempted if it would compromise vital structures.