[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 .

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.

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.

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 .

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

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

[ ABSTRACT] AIM:To observe the effects of total saponins of Panax ginseng ( TSPG) on the promotion of hema-topoiesis and to explore the underlying mechanism in treating aplastic anemia ( AA) in a mouse model.METHODS:For preparation of AA model, BALB/c mice were exposed to sublethal dose of 5.0 Gyγradiation, followed by transplantation of lymphocytes from DBA/2 donor mice.The experiment was divided into 6 groups, including normal control group, AA model group, TSPG treatment groups with low, medium and high doses, and positive control group with cyclosporine A ( CsA) .Both TSPG and CsA were administered by gastrogavage.After 15 d treatment, the peripheral hemogram was test-ed, the cytokine contents in serum were measured, and bone marrow semisolid culture of colony-forming assay was conduc-ted for determining hematopoietic progenitor cells.The protein levels of extracellular signal-regulated kinase 1/2 ( ERK1/2) and its phosphorylation status in the bone marrow cells were determined.RESULTS:Curative effect of TSPG in trea-ting AA mice was satisfactory.Peripheral counts of white blood cells and platelet, and concentration of hemoglobin in TSPG groups with medium and high doses were significantly higher than those in model control.TSPG increased the colony num-bers of CFU-E, BFU-E, CFU-GM and CFU-MK derived from hematopoietic progenitor cells.Meanwhile, up-regulation of the phosphorylated ERK1/2, decreased contents of Th1 cytokines and increased contents of Th2 cytokines in the serum were observed.CONCLUSION:TSPG possess the dual efficacies on the promotion of hematopoiesis and immunoregulation in AA mice by regulating the expression of Th1/Th2/Th17 cytokines and increasing the phosphorylation of ERK1/2.

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.

Aim To observe the effects of panaxdiol saponins component ( PDS-C) extracted and isolated
from Chinese ginseng herb as new Chinese patent med-icine on the promotion of hematopoiesis and the regula-tion of the immune system in treating mice models with aplastic anemia ( AA ) . Methods For preparation of immune mediated AA models, BALB/c mice were ex-posed to sublethal doses of 5. 0 Gy γ radiation, fol-lowed by transplanted lymphocytes from DBA/2 donor mice. The mice models were divided into six groups in-cluding normal control, AA model, PDS-C treated groups with lower, medium and higher dosages, cy-closporine ( CsA) as positive drug control. Both PDS-C and CsA were administered by gastrogavage for 15 days. The peripheral blood cells counts and bone mar-row pathological examination were tested, the percenta-ges of Th1/Th2/Treg cells from spleen were measured, the protein expression levels of T-bet, GATA-3 and FOXP3 transcription factors in spleen cells were detec-ted. Results Curative effect of PDS-C on treating AA
mice was satisfactory. The peripheral hemoglobin, white blood cells and platelet counts in PDS-C groups with medium and higher doses were significantly higher than those in model control. Meanwhile, PDS-C ele-vated the percentages of Th2 cells and Treg cells, but decreased the percentage of Th1 cells, as well as up-regulated the GATA-3 , FOXP3 and down-regulated the T-bet protein levels. Conclusion PDS-C possesses the activities of promoting hematopoiesis obviously. It can improve marrow myelosuppression, enhance the re-covery of hematopoiestic function, and elevate the pe-ripheral blood cells counts. PDS-C also pays its immu-noregulatory efficacy though recovering from unbal-anced Th1/Th2/Treg cells in treating immune media-ted AA mice.

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.