2.Cloning, expression and purification of inhA from mycobacterium tuberculosis
Shu CHEN ; Wenhong ZHANG ; Chaoneng JI
Chinese Journal of Infectious Diseases 2001;0(03):-
Objective To clone and express inhA gene from mycobacterium tuberculosis , and purify the inhA protein. Methods Recombinant plasmid pET 24b/inhA was constructed and transferred into Escherichia coli . After restriction enzyme analysis and sequencing, the host bacteria were induced by IPTG and the product was identified by SDS PAGE. Furthermore, the overexpressed inhA protein was purified by Nit NTA Superflow system. Results The inhA gene was overexpressed in E. coli, the production was corresponding to 30 percent of total cell protein. Using Nit NTA Superflow,we can get more than 99% purified protein. Conclusions The cloning, expression and purification of inhA gene are successful.
3.Analysis of Hepatic Encephalopathy After TIPSS
Wenhong CHEN ; Yuxin NONG ; Yangsheng LIU
Journal of Chinese Physician 2001;0(01):-
Objective To investigate the causes and characterization of hepatic encephalopathy after transjugular intrahepatic portosystemic shunts(TIPSS).Methods 32 cases of patients with recepted TIPSS operation were enrolled in retrospective study.Results 9 of 32 patients suffered from hepatic encephalopathy were detected out after TIPSS. The present of hepatic encephalopathy was associated with the flow direction of blood in portvein after the operation(P
4.Establishment of a stable gastric cancer cell line with lentivirus-mediated RNA interference for I2PP2A
Hairong SHI ; Ying CHEN ; Changlei LI ; Wenhong QIU
China Oncology 2015;(5):352-359
Background and purpose:Overexpression of inhibitor of protein phosphatase 2 A-2 (I2PP2A) in many tumors including gastric cancer suggests that I2PP2A may contribute to the development of gastric cancer. To further study the biological function of I2PP2A and its role in gastric cancer, we established a BGC823 cell line for stable expression of shRNA targeting human I2PP2A gene. Methods: A double-stranded shRNA targeting the I2PP2A was designed, synthesized and was inserted into a lentivirus vector (pGLV2), and the insertion was identiifed by restriction endonuclease analysis and DNA sequencing. BGC823 cells were then transfected with the packaged recombinant lentivirus, and resistant cell clones were selected with puromycin. The expression of I2PP2A was examined using real-time PCR (RT-PCR) and Western blot. Results:Sequencing result proved that recombinant lentivirus vector pGLV2-shRNA-I2PP2A was constructed correctly. RT-PCR and Western blot results conifrmed that the expression of I2PP2A was signiifcantly down-regulated in this infected BGC823 cell line. The efifciency of siRNA interference of I2PP2A could be up to about 90%. Conclusion:A lentiviral vector carrying a shRNA targeting the I2PP2A gene is successfully constructed, and a BGC823 cell line stably expressing I2PP2A shRNA is established with this lentiviral system.
5.Analysis of the relative factors affecting the functional restoration of knee joint after the fracture of patella
Guoxin WANG ; Zhongli JIANG ; Tao LI ; Wenhong CHEN ;
Chinese Journal of Tissue Engineering Research 2002;6(2):290-291
Objective To explore the relative factors affecting the functional restoration of knee joint after the fracture of patella, and to anticipate the effect of middle term and short time rehabilitation therapy. Method 23 patients with limited flexion function of knee joint after the fracture of patella, accepted the routine rehabilitation treatment in out patient service, and some of the patients adopted local heat therapy at the same time. The 13 factors have been selected for regression analysis step by step under the help of SPSS statistical software. Result The flexible degree of the knee joint before rehabilitation (X1), the non smooth posterior edge of patella (X2), and the immobilization time of the joint (X3) are relative factors for the flexible degree of knee joint 1 month after ehabilitation (Y1) and the flexible degree of knee joint 3 onths after rehabilitation (Y2), and Y1 can also be affected by whether surgery therapy or not (X4), (partial regression coefficient P< 0.05). The differences between theoretical mean and practical mean of Y1 and Y2 are 5.6% and 4.2% respectively in the 23 cases. Conclusion The flexible degree of the knee joint before rehabilitation, whether the posterior edge of patella is smooth or not, and the immobilization time of the joint are relative factors affecting the middle term and short term flexible function of knee joint after the fracture of patella, and short term restoration can also be affected by whether surgery therapy or not. The effects of middle term and short term rehabilitation can be predicted by regression equation primarily.
6.Study on extracting process of Fumai Decoction
Wenhong LI ; Riyue YU ; Qi CHEN ; Liang WANG ;
Chinese Traditional and Herbal Drugs 1994;0(10):-
Object To optimize the extraction conditions of Fumai Decoction. Methods Uniform design test was used, and the contents of glycyrrhizinic acid and ginsenoside Rb 1 in extracted solvent were determined by HPLC. Results The ethanol volume and the extracting temperature in the uniform test impacted the test remakably. The best extraction process of Fumai Decoction as follows: the raw drugs were extracted by 10 times of 50% ethanol at 60 ℃ for 4 hours. Conclusion Glycyrrhetinic acid and ginsenoside Rb 1 contents in Fumai Decoction are higher according to the above extraction process.
7.PROBLEMS WORTH EMPHASIS IN ELDERLY RENAL TRANSPLANT RECIPIENTS
Zixuan CHEN ; Wenhong LIN ; Ming ZHAO ; Liangming OU ; Rongxi LIN
Medical Journal of Chinese People's Liberation Army 1982;0(03):-
This paper reported some problems in relation to the elderly renal transplant recipients.Before transplantation,hemodialysis,blood transfusion,water and electrolyte balance and corrections of acidosis.and treatment of diseases of the other systems should be emphasized.Method of treating the athe erotic change in the iliacartery during the operation,and selection of immunosuppressants and dosages were discussed.The prevention and treatment of the postoperative pulmonary infection,septicemia,steroid induct ulcerations of gestrointestinal tract and profound hemorrhage,serious diabetes mellitus and hypertensive encephalopathy after transplantation were also discussed in this paper.
8.Correlation of lipocalin-2 and its receptor expressions with preeclampsia
Wenhong CAO ; Weiping CHEN ; Jing LI ; Yuanhua YE
Chinese Journal of Obstetrics and Gynecology 2012;47(6):418-421
Objective To research the correlation of the expressions of lipocalin-2 (LCN-2) and its receptor (NGALR) in serum and placenta with preeclampsia.Methods From Dec.2010 to Apr.2011,64 women with preeclampsia who delivered in Affiliated Hospital of Qingdao University Medical College were recruited in the study,including 26 women with moderate preeclampsia ( MPE group) and 38 women with severe preeclampsia (SPE group).Twenty-five healthy pregnant women were taken as control group.LCN-2 and NGALR mRNA and protein expression in placenta were measured by reverse transcription-PCR ( RTPCR) and western blot,respectively.Results ( 1 ) The serum levels of LCN-2 in MPE group and SPE group [ (58 ±20),(90 ± 18) μg/L] were significantly higher than that in control group [ ( 19 ±6) μg/L,P<0.01] ; the serum LCN-2 level in SPE group was significantly higher than that in MPE group (P <0.01).(2) LCN-2 mRNA expression in placenta in MPE group and SPE group (0.55 ±0.14,0.61 ±0.14) were both significantly higher than that in control group (0.28 ±0.16,P <0.01 ) ; LCN-2 protein expression in placenta of MPE group and SPE group ( 2.2 ± 0.4,2.4 ± 0.5 ) were also significantly higher than that in control group (1.4 ±0.4,P <0,01 ),no significant difference was found between MPE group and SPE group ( P > 0.05 ),(3) No significant difference was found in the expressions of NGALR mRNA in placenta among MPE group,SPE group and control group (0.46 ±0.1l,0.46 ±0.14,0.45 ±0.15,P >0.05 ).(4) NGALR protein expressions in MPE group,SPE group and control group were 2.7 ±0.8,3.0 ±0.9,and 2.7 ± 0.9,and there were no significant difference among these three groups ( P > 0.05 ).(5) In preeclampsia,serum LCN-2 level significant associated with 24 hours total urinary protein and uric acid ( r =0.565,0.476,P<0.01).LCN-2 serum level were not associated with systolic pressure and diastolic pressure (P > 0.05 ) ; there were no association with the expressions LCN-2 mRNA aud protein in placenta ( P > 0.05).Conclusions Serum LCN-2 level is closely related to the progress of preeclampsia.Increasing expression of LCN-2 in placenta may be a compensatory response to preeclampsia.
9.Neuronal protection of progesterone against ischemic brain injury and underlying molecular mechanisms
Yanying ZENG ; Wenhong ZHI ; Xinsheng DING ; Zheng WANG ; Weixian CHEN
Chinese Journal of Neurology 2013;(6):387-391
Objective To investigate the effect of progesterone pretreatment of focal cerebral ischemic and reperfusion injury (fCIRI) and underlying molecular mechanisms.Methods A single intraperitoneal injection of progesterone (8 mg/kg) given 1 h,48 h and 96 h before fCIRI was established in male Sprague-Dawley rats.The number of survival of neurons in hippocampal CA1 region of the ischemiaside,as well as spatial memory function,was detected on days 3-8 after fCIRI.Extracellular-signalregulated kinase 1/2 phosphorylation (p-ERK1/2) and nuclear translocation of p-ERK1/2 in hippocampal CA1 region were examined using western blot.Results The number of survival of neuronal cells was significantly increased in ischemic groups treated with progesterone at 1 h and 48 h pre-fCIRI (164.3 ± 11.0,218.5 ± 9.1 and 142.7 ± 12.1,F =29.4,P < 0.01) compared with fCIRI group treated with vehicle.Likewise,the escape-latency to reach the hidden-platform recorded in day 5 of Morris water maze test was reduced markedly in fCIRI-treatment groups compared with the vehicle group(10.3 ± 11.1,19.2 ±9.6 and 32.4 ± 14.3 ;F =35.8,P <0.01).The level of p-ERK1/2 was elevated notably during 24 h to 48 h postprogesterone by western blot,while restored to the baseline at 96 h post-progesterone.Improved nuclear translocation of p-ERK1/2 was observed from 2 h to 48 h post-progesterone.The progesterone receptor antagonist RU486 blocked the exaltation of either intracellular level or nuclear translocation of p-ERK1/2,which was induced by progesterone.Conclusions The pretreatment with progesterone exerts a neuroprotective effect against the ischemia-induced neuronal death and ameliorates the deficits in spatial memory through enhancing the activation of ERK1/2.The neuroprotection derived from pretreatment with progesterone achieves a time window of not less than 48 h,which is progesterone receptor-mediated ERK1/2 signaling pathway-dependent.
10.Extraction of diosgenin from Dioscorea zingiberensis
Feng HAN ; Wenhong LI ; Dong LI ; Xuan TANG ; Lianhong CHEN
Chinese Traditional Patent Medicine 1992;0(12):-
AIM:To study the application of ultrasonic extraction in process of extracting diosgenin from Dioscorea zingiberensis C.H.Wright,and to evaluate its merits.METHODS:A new process of ethanol extracting and hydrochloric acid hydrolysis was provided to extraction diosgenin,and five different extraction methods were studied and compared,and the reaction mechanism was discussed.RESULTS:The ultrasonic extraction method was more efficient.Compared with the traditional water-hydrolylis,extraction rate was 15 percent higher,dosage of hydrochloric acid was registered 90 percent decrease,and the production was purer.CONCLUSION:The grinding pretreatment of Dioscorea zingiberensis is help to enhance the effects of ultrasonic extraction,the method has some advantages in efficiency,energy-saving and low pollution.