1.Construction of a C3aR expres sion vector and infection in tubular epithelial cells
Jingmin ZHENG ; Guang YIN ; Wenjin ZHAO ; Zhihong LIU
Journal of Medical Postgraduates 2015;(4):355-359
Objective Our previous study showed that the expression of C 3aR was increased in renal tubular epithelial cells (RTEC).However, the role of C3aR in diabetic nephropathy remains unknown, and so does the exact physiological and pathological signifi-cance of C3aR in relevant renal tissues.In the present study, we investigated the physiological and pathological significance of C3aR in RTECs using a lentivirus expression vector and anRTEC strain overexpressing C3aR. Methods Based on the sequence of C3aR mRNA, the human C3aR gene was synthesized and cloned into the multi-clonal site of the lentivirus expression vector pLenti6.3-MCS-IRES2-EGFP to construct a C3aR expression vector pLenti6.3-C3aR-IRES2-EGFP.After identification by sequencing, the recombinant lentivirus expressing C3aR was packaged by cotransfecting293T cellswith the recombinant lentivirusexpression vector and packaged plasmid .Then,the recombinant lentivirus was used to infect the human RTECline HK 2.After screening in the medium with blasticidin, blasticidinr-esistant cell clones were obtained, followed by identification ofthe human RTECline stably overexpressing C 3aR by real-time PCR and immunochemical staining. Results TheC3aR expression vector pLenti6.3-C3aR-IRES2-EGFP was successfully constructed and the sequence was proved to be correct. C3aR expression recombinant lentivirus was successfully packaged with a titer of 5×108/mL.Blasticidin-resistant HK2 cell clones were ob-tained and the expression of HK2-C3aR mRNA was significantly higherin the HK2-C3aR cells than in the non-transfected HK2 cells (2.33± 0.45 sv 1.00±0.09, P<0.05). Conclusion We successfully constructed a C3aR expression lentivirus vector pLenti6.3-C3aR-IRES 2-EGFP and a C3aR overexpression renal tubular epithelial cell strain HK2-C3aR, which is very contributive to further studies of the roles of C3aR in renal tubular epithelial cells and other types of cells.
2.Expression and clinical significance of ABCG2 protein in hepatocellular carcinoma
Dawei ZHAO ; Xiaoyu YIN ; Jinfang ZHENG ; Lijian LIANG
Chinese Journal of Digestive Surgery 2010;9(3):213-215
Objective To investigate the expression and clinical significance of ABCG2 protein in hepato-cellular carcinoma (HCC). Methods Specimens of HCC were collected at The First Aifiliated Hospital of Sun Yat-sen University from January 2005 to December 2006. The expression of ABCG2 protein in 165 samples of HCC tissue, 25 samples of normal liver tissue and 40 samples of cirrhotic liver tissue was detected using immunohisto-chemistry. The correlation between the expression of ABCG2 protein and clinicopathological characters was then analyzed. Enumeration data, survival rate and the difference between groups were analyzed with a chi-square test, the Kaplan-Meier method and Log-rank test, respectively. Results ABCG2 protein expression was weakly posi-tive in all normal and cirrhotic liver tissues. In HCC tissues, the expression of ABCG2 protein was strongly positive in 66 cases and weakly positive in 99 cases. The expression of ABCG2 protein was related to tumor diameter, tumor number, adjacent organ invasion and TNM stages (χ2 =8. 130, 14. 279, 4. 820, 21. 179, P <0. 05). Kaplan-Meier survival analysis revealed that patients with strongly positive ABCG2 protein had a significantly lower 3-year overall survival (24. 1%) compared with those with weakly positive ABCG2 protein (39. 4%) (χ2 = 15.716, P<0.05). Conclusions The expression level of ABCG2 protein is related to tumor invasiveness, TNM stage and prognosis. ABCG2 has the potential to become a new target for HCC treatment.
3.Effect of decitabine on proliferation and differentiation of K562 cells
Xiaoling YU ; Yanna ZHAO ; Zhiyin ZHENG ; Ruilan GAO ; Liming YIN
Chinese Journal of Pathophysiology 2017;33(1):13-17
AIM:To investigate the effect of decitabine (Dacogen, DAC) on the proliferation and differentia-tion of K562 cells.METHODS:The K562 cells were treated with different concentrations of DAC .The colony formation ability of the cells was detected by the colony formation assay with semi-solid culture .The cell viability was detected with MTT assay.The morphologic features were observed under inverted microscope with Wright ’s staining.The changes of the cell cycle distribution and the expression of CD 11b and CD42b were analyzed with flow cytometry .The protein expression of CDK2, cyclin E1, P27, GATA-1 and PU.1 in the K562 cells was determined by Western blot .RESULTS:DAC signifi-cantly decreased the colony number of the cells and cell viability in a dose-dependent manner .The morphological changes of the cells displayed partial differentiation .After treated the K562 cells with DAC for 72 h, the cell proportion in S phase was obviously decreased , while the cell proportion in G 2/M phase was obviously increased in a dose-dependent manner . After treated the K562 cells with DAC for 7 d, the percentage of CD11b and CD14 positive cells was further elevated , and the protein expression of P27, GATA-1 and PU.1 was increased.However, the protein expression of CDK2 and cyclin E1 was decreased .CONCLUSION:DAC inhibits the proliferation and induces differentiation of the K 562 cells via regulation of cell cycle .
4.Construction of C3a secrectory expression vector and tubular epithelial cell line stably expressing C3a
Jingmin ZHENG ; Guang YIN ; Shifeng YUN ; Wenjin ZHAO
Journal of Medical Postgraduates 2015;(6):584-589
[Abstract ] Objective Evidence from previous studies indicated that over-activation of C3a/C3aR axis existed in the renal tubular epithelial cells of patients with renal diseases including diabetic nephropathy .However , the pathological significance of over-ac-tivating C3a/C3aR axis still remains to be elucidated .In this study, we constructed a renal tubular epithelial cell line over-expressing C3a in a secretory manner in order to provide a cell model to investigate the pathological significance of over -activating C3a/C3aR axis under various pathological scenes . Methods We designed a synthesized C3a secretory expression unit and cloned it into the multi-clonal site of lentivirus expression vector pLenti 6.3-MCS-IRES2-EGFP.After identification by sequencing , recombinant lentivirus was packaged by using pLenti 6.3-C3a-IRES2-EGFP and packaging plas-mid in 293T cells.Then, the recombinant lentivirus was used to in-fect HK2, a cell line of human renal tubular epithelial cells .After screening in medium with blasticidin , blasticidin resistant cell clones were obtained .Real-time PCR and ELISA method were applied to analyze the expression and secretion of stable transfected cells cloned C3a and identify renal tubular epithelial cell lines with stable over-activating C3a. Results ①C3a secretory expression unit was suc-cessfully synthesized and correctly cloned into the multi-clonal site of pLenti6.3-IRES2-EGFP; ②C3a secrectory expression recombi-nant lentivirus LV-C3a was successfully packaged with a high titer of 5 ×108/mL;③HK2 Cell clones resistant for blasticidin were ob-tained;according to the analysis of Real-time PCR and ELISA, the C3a mRNA level in HK2-C3a cell lines was significantly higher than that of HK2 cells(1.0 ±0.5 vs 1321.0 ±18.0, P<0.01) and the secreted C3a level increased significantly ([0.3 ±0.2]ng/mL vs [249.0 ±37.0] ng/mL, P<0.01). Conclusion The present study successfully constructed C 3a secretory expression vector pLenti6.3-C3a-IRES2-EGFP and C3a over-expression renal tubular epithelial cell line HK 2-C3a, which is very useful in further study of the function and significance of C 3a/C3aR axis not only in renal tubular epithelial cells but also in other cell types .
5.Classification and operation in the treatment of maxillary retrusion of adult patients with cleft lip and palate.
Yilue ZHENG ; Ningbei YIN ; Zhenmin ZHAO ; Xiaomei SUN ; Chanyuan JIANG ; Haizhou TONG ; Hengyuan MA ; Tao SONG
Chinese Journal of Plastic Surgery 2016;32(1):3-8
OBJECTIVETo classify the patients with cleft lip and palate who need orthognathic surgery and to propose the corresponding operations.
METHODSFrom January 2005 to May 2015, 121 patients with cleft lip and palate diagnosed as maxillary retrusion were treated by orthognathic surgery. Inclusion criteriar: (1) male aged over 16, female aged over 14; (2) diagnosed as non-syndromic cleft lip and palate without systemic disease and other genetic diseases; (3) without previous orthodontic and orthognathic treatment; (4) having no other craniofacial malformation. Maxillary features and repaired types were recorded.
RESULTS93 patients were included and divided into two categories depended on the dental crowding. Class I: the teeth quantity and bone quantity is coordinated, space analysis ≤ 4 mm (mild dental crowding). The forward distance of maxillary less than 6 mm was defined as Class I a (36 cases) more than 6 mm as Class I b (28 cases). Class II: the teeth quantity and bone quantity is not coordinated, space analysis > 4 mm ( moderate or severe dental crowding). After the simulation of distraction osteogenesis, the anterior crossbite was corrected defined as Class II a (23 cases), not corrected defined as Class II b (6 cases). Class I a were corrected by conventional orthognathic surgery. While Class I b were corrected by Le Fort I maxillary advancement using distraction osteogenesis. Class II a were repaired just by anterior maxillary distraction. While Class II b need to combine conventional orthognathic surgery with anterior maxillary distraction. All the patients were satisfied with the treatment effect.
CONCLUSIONSThe patients of cleft lip and palate with maxillary retrusion who need orthognathic surgery can be classified as the method mentioned above, and then choose the appropriate operations.
Adolescent ; Adult ; Cleft Lip ; complications ; Cleft Palate ; complications ; Female ; Humans ; Male ; Maxilla ; Osteogenesis, Distraction ; Osteotomy, Le Fort ; Retrognathia ; classification ; surgery
6.Correlative factors on the articulation disorder of patients with cleft palate.
Chunli GUO ; Li MA ; Heng YIN ; Yang LI ; Shufan ZHAO ; Bing SHI ; Qian ZHENG
West China Journal of Stomatology 2014;32(5):432-435
OBJECTIVETo investigate the correlation between articulation, velopharyngeal function, and surgical age by comparing the changes in articulation after velopharyngeal closure is performed. This study is also conducted to investigate the influencing factors of omission change between pre- and post-operation.
METHODSA total of 48 patients, including 18 males and 30 females, mean age (13.3 ± 5.8) years, with non-syndromic cleft lips and palates were selected from January 2011 to December 2011. Their speech data and articulation between pre- and post-operation were retrospectively analyzed using non-parametric tests. Correlation study was performed to analyze the influencing factors of the changes in articulation. P < 0.05 was considered statistically significant.
RESULTSThe difference in articulation after velopharyngeal closure occurred was significant (Z = -3.796, P = 0.000). A negative correlation between the ratio of post-operative normal articulation and surgical age (R = -0.487, P = 0.000) was observed. The change in omission was positively correlated with surgical age (R = 0.589, P = 0.000) and gender (R = 0.404, P = 0.047). By comparison, the change in omission was negatively correlated with follow-up time (R = -0.235, P = 0.040).
CONCLUSIONArticulation and intelligibility are significantly improved after velopharyngeal closure is performed. These parameters are negatively correlated with surgical age to some extent. In addition, the change in omission is positively correlated with surgical age and gender, whereas the change in omission is negatively correlated with follow-up time.
Adolescent ; Articulation Disorders ; Child ; Cleft Lip ; Cleft Palate ; Female ; Humans ; Male ; Retrospective Studies ; Velopharyngeal Insufficiency ; Young Adult
7.Progress on roles and mechanisms of farnesoid X receptor (FXR) in chronic liver diseases
Wenxuan XU ; Zili ZHANG ; Shifeng ZHAO ; Feng ZHANG ; Li WU ; Yin LU ; Shizhong ZHENG
Chinese Pharmacological Bulletin 2016;32(3):314-318,319
Farnesoid X receptor ( FXR) plays a key role in me-tabolism of substance, such as bile acid, lipid, glucose,( etc) . Newly published credible discoveries have claimed that as a reg-ulatory hub in metabolism, FXR is closely linked with diverse chronic liver diseases, including viral hepatitis, alcoholic fatty liver disease, nonalcoholic fatty liver disease, hepatic fibrosis and hepatocellular carcinoma. This review summarizes the roles and mechanisms of FXR during the courses of chronic liver dis-eases, aiming at providing novel insights and therapeutic target for antifibrotic research and drug development.
8.Herb-herb interaction in traditional Chinese medicine:a review of pharmacokinetics-based interaction in Herb-Pairs
QIANG WEI-JIE ; CHEN YING ; ZHAO ZHENG ; YIN JIE ; ZHU XIAO-XIN
Chinese Journal of Pharmacology and Toxicology 2017;31(10):996-996
″Herb-Pairs″, also known as pair drugs, refers to a prescription consisted of two relatively fixed traditional Chinese medicine, is the most basic, most simple and most common form of medication prescription in traditional Chinese medicine compound compatibility. It is not a random combination of two herbs, nor is the simple accumulation of efficacy, but the simple and delicate experience of ancient Chinese medicine practitioners. As a bridge between single drug and prescriptions, it is the embodiment of the regular and dialectical connotation. Therefore, research on Herb-Pairs has always been the most basic and most important entry point for compound compatibility studies. However, the interaction between herbs and herbs is an effect with a downside as well as benefits. The beneficial herb-herb interaction in Herb-Pairs include mutual promotion, mutual enhancement, mutual restraint between two drugs and counteract toxicity of another drug. And the harmful herb- herb interaction in Herb- Pairs includes mutual inhibition and antagonism. All of these interactions areby means of affecting the metabolism of components to play a therapeutic effect. Using the pharmacokinetic-pharmacodynamic (PK-PD) binding model, the combination of drug metabolism and pharmacodynamics can further elucidate the influence on effect caused by drug concentration and metabolism, which can help elucidate the mechanism of drug action. Consequently, in this review, the herb-herb interactions in terms of pharmacokinetic were summarized to elucidate rule of TCM compatibility.
9.Qualitative research on the demands of nursing staff on nursing repository
Wei WANG ; Xiaoping ZHAO ; Xiaohong YIN ; Xiaohong ZHENG ; Qun JIN ; Xiaolian ZHANG
Chinese Journal of Nursing 2009;44(8):742-744
Objective To explore the demands of nursing staff on nursing repository and provide reference for the develop-ment of nursing repository. Methods In-depth interview was conducted on 21 nursing staff by qualitative research. The themes were formed by category analysis. Results There were four themes about the demands of nursing staff on nursing repository:necessity to develop nursing repository ,contents of the repository ,forms of the repository and prospect of the reposi-tory. Conclusions Nursing staff need a nursing repository. They hope that the repository can provide comprehensive,concrete and practical knowledge,and provide a good interface with digitization. The design of repository should be consistent with in-ternational standards.
10.Comparison of different anode filter combination with full-field digital mammography: imaging quality versus radiation dose
Yongxia ZHAO ; Jingwen WANG ; Xiaohong YANG ; Qiuping LI ; Xiaoping YIN ; Jian SUN ; Yating ZHENG
Chinese Journal of Radiology 2012;46(10):929-931
Objective To compare imaging quality and radiation dose of different anode filter combination for full-field digital mammography.MethodsThe image of FLUKE NA 18-220 phantom were taken at full-field digital mammography (FFDM),system with Mo/Mo,Mo/Rh,Rh/Rh anode/filter combination by automatic exposure control,record the exposure factors and doses.The images on monitor with the best window width and window level were read by 4 independent radiologists,the images of specks groups,nylon fibers and masses was assessed by the 4 experienced readers at the criterion of American College of Radiology 1999 mammography quality control manual.Statistical analysis was performed using analysis of variance.ResultsThe nylon fibers scores of Mo/Mo,Mo/Rh and Rh/Rh were 5.50 ± 2.12,4.50 ± 1.85 and 4.38 ± 1.38 ; the specks groups scores of Mo/Mo,Mo/Rh and Rh/Rh were 5.38 ± 1.98,4.25 ± 1.56 and 4.38 ± 1.38;the masses scores of Mo/Mo,Mo/Rh and Rh/Rh were 5.38 ± 1.98,4.38 ±1.68 and 4.25 ± 1.56,the detection of specks groups,nylon fibers was not statistically significant ( F =4.56 and 4.32,P > 0.05 ),but the detection of masses was statistically significant ( F =36.65,P < 0.05).The radiation doses were different,the entrance surface dose (ESD) and average glandular dose (AGD) of Rh/Rh were (5.11 ± 1.89) and ( 1.08 ± 0.13 ) mGy,the ESD and AGD of Mo/Mo were (6.66 ± 2.33 )and ( 1.29 ± 0.38 ) mGy,the ESD and AGD of Mo/Rh anode/filter combination were (5.67 ± 2.02) and ( 1.29 ± 0.38) mGy.ConclusionsThe radiation dose of Rh/Rh and Mo/Rh anode/filter combination of FFDM were lower,and the imaging was clear,so Rh/Rh and Mo/Rh anode/filter combination of FFDM prefer to mostly patients,especially when the thinkness is large,Rh/Rh anode/filter combination is preferred,because the kV values was higher,the penetration of X-ray was stronger.The Mo/Mo anode/filter combination was used when the needs of high-resolution,because the detection of masses of it was better.