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.New multiplex allele-specific PCR assay targeting katG codon 315 variation for detection of isoniazid resistant Mycobacterium tuberculosis strains
Jialin JIN ; Wenhong ZHANG ; Xinhua WENG ; Shu CHEN ; Lingyun SHAO
Chinese Journal of Laboratory Medicine 2003;0(08):-
Objective To develop a new multiplex allele-specific PCR(MAS-PCR) assay to detect mutation in codon 315 of katG gene of Mycobacterium tuberculosis , mutation in this codon has been reported to be able to account for the Mycobacterium tuberculosis clinical isolates resistant to isoniazid(INH).Method Based on the sequence of katG gene of Mycobacterium tuberculosis , three specific primers are designed to carry out the MAS-PCR, 84 purified DNA preparation with known katG 315 variation detected by PCR-restriction fragment length polymorphism (PCR-RFLP) are used to optimize PCR.Results 84 Mycobacterium tuberculosis clinical stains are detected by the MAS-PCR and PCR-RFLP, respectively.The sensitivity of detection by MAS-PCR is 77.8%,and the specificity is 95.2%.katG mutation S315N(AGC→AAC), neglected in RFLP, can be detected by MAS-PCR.Conclusion MAS-PCR assay is sensitive, specific, economic and easy to carry out , can be used in clinical laboratories to detect the INH-resistant Mycobacterium tuberculosis strains.
6.Experimental Study of Changyanqing for Colitis Induced by Dextran Sulfate Sodium
Yonghui LU ; Wenhong CHEN ; Pinjin HU ; Sheng HU
Journal of Guangzhou University of Traditional Chinese Medicine 1999;0(02):-
[Objective] To observe the effects of Changyanqing, a Chinese prescription with the actions of regulating qi and activating blood, strengthening spleen-qi and clearing away heat, on ulcerative colitis (UC) and to explore its therapeutic mechanism . [ Methods ] Forty mice were randomized into Changyanqing group ( Group A ), salicylazosulfapyridine group (Group B), Changyanqing and salicylazosulfapyridine (Group C) and model group (Group D). Mouse models with UC were induced by dextran sulfate sodium (DSS). Effects of Changyanqing on disease activity index (DAI) and intestinal myeloperoxidase (MPO) content were observed. [Results] Changyanqing reduced DAI and MPO activity and its effect was similar to that of salicylazosulfapyridine. [ Conclusion ] Changyanqing exerts a better effect in treating DSS-induced UC and one of its therapeutic mechanisms may be related to the reduction of MPO activity.
7.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.
8.Secondary pathological changes in infant with developmental dysplasia of the hip: the application value of high frequency ultrasound
Wenhong YI ; Hongmei LIU ; Sushu LI ; Yanni HE ; Yanjun CHEN
Chinese Journal of Ultrasonography 2016;25(7):612-615
Objective To investigate the clinical value of high frequency ultrasound in discovering secondary pathological changes in infant with developmental dysplasia of the hip (DDH).Methods Thirtyfour infants with 36 dislocation hips were involved.Among them,32 were with unilateral dislocation of hip joint,2 were with bilateral dislocation of hip joint.High frequency ultrasound were used to observe the morphological changes of hip glenoid labrum,adipose hyperplasia in acetabulum,thickness variation of articular capsule,and development of the ossification center of the femoral head.Besides,the morphological changes of hip glenoid labrum and adipose hyperplasia in acetabul were compared with those from MRI.Results The sensitivity and specificity of ultrasound in diagnosing glenoid labral introversion were 82 % and 100% respectively.The coincidence rate of diagnosing glenoid labral introversion and glenoid labral extroversion were both 83%.Adipose hyperplasias in acetabul were clearly seen in 31 hips using ultrasound.The thickness of articular capsule in the affected hip and in the healthy hip was (2.8 ± 0.6)mm versus (1.7 ± 0.4) mm,and the difference was obviously significant (P <0.01).The ossification center of the femoral head in the 34 infants were classified as the following four types:① which was smaller in the affected side than in the healthy side (22/34);②which was shown in the healthy side (7/34) but not in the affected side;③ which in both sides were not shown (4/34);④ which in both sides were small (1/34).Conclusions Ultrasound is a method of simple,rapid,non-invasive in diagnosing DDH,and it can be used to discover secondary pathological changes in infant with DDH,which provide important reference for clinical diagnosis and treatment.
9.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.
10."Construction of ""STAR"" nurse training model and evaluation of its application effect"
Wei WANG ; Wenhong ZHOU ; Siqin DONG ; Chang′an LI ; Lin CHEN ;
Chinese Journal of Practical Nursing 2016;32(32):2481-2485
Objective To construct and implement theSTARnurse training model, and discuss its application effect and the problems that should be paid attention to, and to provide operational cases and practical basis for nursing clinical education. Methods Through literature review and expert consultation, the framework and content of STAR nurse training model were set up and implemented. The questionnaire survey and semi structured in-depth interviews were conducted among the nurses in the hospital to evaluate the effect of improving the nurses′self-directed learning ability. Results After the implementation of the project, the scores of the three dimensions of self-management, desire for study and self-control were (3.67±0.57), (4.05±0.54), (3.99±0.50) points, which were higher than (3.55±0.49), (3.71± 0.52), (3.53±0.42) points before implementation. The difference was statistically significant (P<0.05 or 0.01). The semi structured in-depth interviews showed that all the nurses believed that STAR nurse training model could promote independent learning and stimulate interest in learning. 14 nurses thought it was beneficial for the nurses to find the problems. Conclusions STAR nurse training model can create a favorable learning environment for nurses, and stimulate the learning motivation. It plays a positive role in improving nurses′ability of self-directed learning.