[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 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.

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.

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

Objective To study the methods and results of endovascular treatment of posterior inferior cerebellar artery (PICA) aneurysms. Methods Twenty-one patients with PICA aneurysms were treated with endovascular treatment. The locations of aneurysm on PICA were evaluated through the DSA. Eight patients received single coil embolization, 5 received liquid Glue embolization, 2 received coil embolization combined with liquid Glue, 2 received coil embolization assisted with stents, and 4 underwent ocelasion of the parent PICA. Outcome was evaluated with the Glasgow outcome scale (COS). Results There were complete (100%) occlusion in 5 patients, near complete (>90% ) occlusion in 2, and incomplete (85%) occlusion in 1 in single coil embolization. Seven patients with Glue embolization (n = 5 ) or combination with coils ( n = 2 ) exhibited complete ( 100% ) occlusion. There were near complete ( >90% ) occlusion in 2 cases with coil assisted with stents. Complete occlusion of the parent PICA was achieved in 3 patients, and near complete occlusion of PICA in one case. One patient suffered from new neurological deficits, and one patient treated with coils and stents died. None of the patients suffered from re-bleeding. These patients received follow-up during a mean period of (22±8) months. Overall long-term outcome was good ( GOS score 4 or 5 ) in 17 patients, poor ( COS score 2 or 3) in 3, and fatal ( GOS score 1 ) in one case. Conclusions According to the location of aneurysms on PICA, aneurysms can be effectively and safely treated with endovaecular embolization.

Objective To investigate the application value of identification of the scalp surface locations of cerebral ischemia lesions before direct revascularization for moyamoya disease and to design surgical approaches according to this by using the fusion of single photon emission computed tomography ( SPECT) cerebral perfusion imaging with CT imaging. Methods The clinical data of 13 adult patients with ischemic-type moyamoya disease underwent superficial temporal artery-middle cerebral artery bypass surgery were analyzed retrospectively. SPECT cerebral perfusion imaging was fused with CT imaging of the same machine before procedure. The lesions of ischemia were located on the cortical surface. The surgical approaches were designed at the center of the ischemic lesions. The patients were followed up for 6 to 12 months after procedure. The improvement of clinical symptoms and cerebral perfusion of the patients were observed after operation. Results One patient had perioperative cerebral hyperperfusion syndrome,and the others did not have any perioperative complications. At one-month follow-up, the improvement of symptoms in 4 patients were excellent,in 5 were good,in 4 were fair,and none was poor. At 6 to 12 month follow-up,the improvement of symptoms in 9 patients were excellent,in 4 were good,and none was poor. The postoperative visual SPECT cerebral perfusion imaging analysis suggested that the cerebral perfusion was improved significantly as compared with before procedure in all patients. Quantitative analysis:There was significant difference in target ischemic lesions between preoperative Fb and postoperative Fb ([2. 13±1. 06]% vs. [4. 13±2. 09]%;P<0. 05). There was significant difference between preoperative Fb and Fe ([2. 46±1. 97]% vs. [2. 13±1. 06]%;P<0. 05). The postoperative BFCR was [67. 57±3. 78]%( >50%) , which indicated that the efficacy of the procedure was remarkable. The superficial temporal arteries fed to brain of the patients were observed after procedure by using the head CT angiography. The postoperative head MRI reexamination showed no new infarcts occurred at 6 months. Conclusion Combine SPECT cerebral perfusion imaging with CT imaging to design surgical approach for superficial temporal artery-middle cerebral artery bypass surgery may improve the efficacy and reduce the risks of operation.

[ 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.

Objective To explore the effective countermeasures for treatment of multiple trauma. Methods The clinical data of 135 cases with multiple trauma admitted into the emergency center of Zhongshan City from June 2002 to June 2003 were applied to study the mode and the key point of integrated treatment for multiple trauma. Results Of 135 cases,107 survived with a success rate of 79.3% and 28 died with a mortality of 20.7%. Conclusions (1)The integrated treatment in the emergency center is an effective way to improve the curative rate of multiple trauma as well as a development trend in treatment of multiple trauma. (2) The mode to make integrated diagnosis and treatment includes the following parts: set up wound center in emergency center,build a united rescue system and train high quality professional personnel. The diagnosis and treatment of multiple trauma must be organized and carried out by doctors in the emergency center. (3)The key treatment points for serious multiple trauma include firstly,active pre-hospital medical care and in-hospital medical care to save the lives;secondly,timely surgical operation to repair the damaged organ and eliminate the factors of lethality and cripples;and thirdly,necessary intensive care to treat the original damage further,protect various function of organs,promote recovery of body and prevent the complications.