Objective To construct five shRNA-expression plasmids and to investigate the expression of Smad2 in TGF-?/ Smads signal transduction treated with shRNA-expression plasmid.Methods Five shRNA-Smad2 DNA sequences from mRNA sequence of mouse Smad2 gene were designed and synthesized.DNA oligonucleotides encoding an appropriate shRNA were inserted to shRNA expression vector respectively.Five shRNA-Smad2 expression plasmids were obtained and then transfected into NIH/3T3 cells.The suppressed expression of Smad2 was assessed by RT-PCR and Western-blotting.Results The shRNA-expression plasmid numbered 2.4 could markedly reduce the expression of Smad2.The suppression effect of the RNAi-pool composed of four different plasmids was more obvious than that of any single.Conclusion The shRNA-expression plasmids were successfully constructed,which could specifically and effectively suppress the expression of Smad2.The method of using a mixture of RNAi plasmids to improve the RNAi efficiency was established.

AlM:To evaluate the expression of transcriptional factor lslet-1 in retina in experimental retinal neovascularization induced by oxygen.
METHODS: The murine retinal neovascularization were induced by hyperoxia exposure. The morphological observation of retinal neovascularization was performed using angiography by fluorescein dextran injection under the fluorescence microscope, and the new blood vessels were quantified after 5d in room air (17-day-old) by counting the vascular epithelial cell nuclei protruding into viteous cavity using HE stain. Realtime PCR and Western blot were used to examine retinal lslet-1 level in postnatal 7,12, 14,17 and 26d respectively.
RESULTS: A lots of new blood vessels were demonstrated in the mouse retina in hyperoxic group by fluorescein angiography and histological method. Moreover, no significant difference was found in retinal lslet-1 level in postnatal 7d between hyperoxic group and control group, but was significantly higher in postnatal 12, 14 and 17d mice compared with control mice. However, mice at postnatal 26d, expression of lslet-1 in retina decreased to normal level.
CONCLUSlON: ln processing mouse model of retinal neovascularization, sustained hypoxia retinal tissue induce retinal neovascularization by increas the expression of transcription factor lslet-1.

Objective:To investigate the influencing factors for the number of lymph node harvested after Da Vinci robotic and laparoscopic radical gastrectomy for gastric cancer.Methods:The retrospective case-control study was conducted. The clinicopathological data of 1 396 patients who underwent Da Vinci robotic or laparoscopic radical gastrectomy for gastric cancer in the First Affiliated Hospital of Nanchang University from December 2014 to July 2019 were collected. There were 991 males and 405 females, aged (60±11) years. Surgery using Da Vinci robotic system or laparoscopic system was completed according to patients' wishes. Cases with early gastric cancer underwent D 1+ lymphadenectomy and cases with advanced gastric cancer underwent standard D 2 lymphadenectomy. Observation indicators: (1) intraoperative situations; (2) postoperative situations; (3) influencing factors for the number of lymph node harvested after radical gastrectomy for gastric cancer; (4) follow-up and survival. Follow-up using outpatient examination or telephone interview was performed to detect survival of patients up to October 2020. Measurement data with normal distribution were represented as Mean± SD. Univariate analysis was done using the chi-square test or Fisher exact probability. Multivariate analysis was performed using Logistic regression model. The survival rate was calculated by Kaplan-Meier method. Results:(1) Intraoperative situations: all the 1 396 patients underwent radical gastrectomy, including 415 cases undergoing Da Vinci robotic radical gastrectomy and 981 cases undergoing laparoscopic radical gastrectomy. Thirty-five of the 1 396 patients were converted to open surgery, including 5 cases undergoing Da Vinci robotic radical gastrectomy and 30 cases undergoing laparoscopic radical gastrectomy. Of the 1 396 patients, 983 cases underwent distal gastrectomy, 400 cases underwent total gastrectomy and 13 cases underwent proximal gastrectomy, among which 597 cases underwent Billroth Ⅰ anastomosis, 385 cases underwent Billroth Ⅱ anastomosis, 401 cases underwent Roux-en-Y anastomosis and 13 cases underwent residual stomach-esophagus anastomosis. The operation time, volume of intraoperative blood loss and cases with intraoperative blood transfusion were (221±51)minutes, (201±81)mL, 24 of 415 cases undergoing Da Vinci robotic radical gastrectomy, and (196±42)minutes, (232±76)mL, 75 of 981 cases undergoing laparoscopic radical gastrectomy, respectively. (2) Postoperative situations: the time to postoperative first flatus, time to postoperative initial liquid food intake and duration of postoperative hospital stay of the 1 396 patients were (3.0±1.0) days, (4.2±1.5) days and (9.0±3.8) days, respectively. Two hundred and ten of the 1 396 patients had postoperative complications including 170 cases with grade Ⅰ-Ⅱ complications and 40 cases with grade Ⅲ-Ⅴ complications. Eight of the 210 patients with postoperative complications died of serious complica-tions and the other 202 cases were cured after symptomatic treatment. Results of postoperative histopathological examination showed that there were 958 cases of adenocarcinoma, 220 cases of mucinous adenocarcinoma, and 218 cases of signet ring cell carcinoma. The number of lymph node harvested and the number of positive lymph node of the 1 396 patients were 26.0±8.3 and 3.6±0.9, respectively, and cases with the number of lymph node harvested ≥16 or <16 were 1 312 and 84. (3) Influencing factors for the number of lymph node harvested after radical gastrectomy for gastric cancer: results of univariate analysis showed that the operating surgeon, operation method, range of gastric resection, nerve invasion, degree of tumor invasion and tumor pathological N stage were related factors influencing the number of lymph node harvested after Da Vinci robotic and laparoscopic radical gastrectomy for gastric cancer ( χ2=13.167, 6.029, 15.686, 5.573, 9.402, 17.139, P<0.05). Results of multivariate analysis showed that the operating surgeon, operation method, range of gastric resection and tumor pathological N stage were independent factors influencing the number of lymph node harvested after Da Vinci robotic and laparoscopic radical gastrectomy for gastric cancer ( odds ratio=1.589, 2.018, 1.787, 0.267, 95% confidence interval as 1.221?2.068, 1.140?3.570, 1.066?2.994, 0.103?0.689, P<0.05). (4) Follow-up and survival: of the 1 396 patients, 1 256 cases were followed up for 2 to 70 months, with a median follow-up time of 27 months. The 3-year cumulative survival rate of the 1 256 cases was 70.2%. Conclusion:The operating surgeon, operation method, range of gastric resection and tumor pathological N stage are independent factors influencing the number of lymph node harvested after Da Vinci robotic and laparoscopic radical gastrectomy for gastric cancer.

Objective:To study the changes of Th17,regulatory T(Treg) cells and IL-17,IL-23 levels at acute phase and recovery phase in children with Henoch-Schonlein purpura(HSP) in order to further understand the immunological pathogenesis and provide help for treating HSP. Methods:The vein blood samples were collected from 65 children with HSP and 30 normal children. The proportion of Th17 cells and regulatory T cells were tested by FCM and concentration of IL-17 and IL-23 in plasma were tested by ELISA. Results:Compared with normal children,the levels of Th17,Th17/Treg and IL-17,IL-23 were in increase at acute phase in children with HSP(P<0. 05). At recovery phase,that were in decrease compared with the acute phase(P<0. 05),but still higher than the normal children(P<0. 05). Compared with normal children,the level of Treg was in decrease at acute phase in children with HSP (P<0. 01). At recovery phase,that was in increase compared with the acute phase(P<0. 01),but still lower than the normal children (P<0. 01). Among the simplex,abdominal and other type of children with HSP,the levels of Th17,Treg,Th17/Treg and IL-17,IL-23 were same( P>0. 05 ) . At acute phase in children with HSP, Th17 cells percentage had positively correlated with IL-17 levels ( r=0. 880,P<0. 01),IL-23 levels had positively correlated with Th17 cells percentage and IL-17 levels (r=0. 838 or 0. 877,P<0. 01). Conclusion:Th17,Treg,Th17/Treg,IL-17 and IL-23 are involved in the course of the immunological pathogenesis in children with HSP,but the levels of that have no significant difference among simplex,abdominal and other types,further researches need to be done.

The composition of S95-100 field mobile medical system was introduced, and some practices were summarized such as rational grouping, enhanced organization, proper allocation and etc. The problems of the system were analyzed from three aspects of the fixation of the internal devices, the rapid deployment in the plateau and maintenance in field conditions, and some measures were put forward to improve the medical support ability of the system in the plateau.

Objective To investigate the p53,Bax,bcl-2 gene in NaAsO2-induced human embryonic lung fibroblasts(HELF)apoptosis.Methods HELF was divided into HELF cells transfected with p53 plasmid(p53 group),HELF cells transfected with PC plasmid(PC group)and normal cultured HELF cells(normal group).The mRNA expression of p53,Bax and bcl-2 gene was detected by real-time PCR,the protein expression of p53,Bax and bcl-2 was assessed by immunohistochemical SABC and the cell apoptosis of HELF was detected by flow cytometry(FCM),in a 6-well plate and cultured for 48 hours,which was exposed to different doses(0,3,9,15mmol/L)NaAsO2 for 24 hours.Results The p53 gene mRNA expression level of p53 group(0.51±0.29)was lower than that of the normal group and PC group [ (1.00 ± 0.20), (1.32 ± 0.26), all P < 0.05 ]. The p53 protein expression level of p53 group(4.10 ± 1.20) was lower than the PC group and normal group[ (8.00 ± 1.63), (7.90 ± 1.79), allP < 0.05]. In p53 group, PC group, normal group exposed to 0,3,9,15 mmol/L NaAsO2 doses, the apoptotic rate [(0.57 ± 0.28)%, (22.91 ± 4.86)%, (40.05 ± 3.93)%, (44.87 ± 3.58)%; (0.65 ± 0.24)%, (14.09 ± 3.49)%,(20.31 ± 3.66)%, (32.42 ± 3.63)%; (0.56 ± 0.25)%, (12.14 ± 3.70)%, (19.61 ± 3.63)%, (30.43 ± 2.83)%], Bax mRNA expression level[(12.73 ± 3.96), (25.12 ± 6.42), (104.96 ± 26.77), (154.04 ± 30.52); (14.63 ± 3.57),(36.75 ± 3.67), (272.26 ± 66.11), (846.12 ± 243.36); (14.75 ± 5.65), (37.22 ± 11.27), (278.51 ± 37.42),(861.67 ± 369.29) ], Bax protein expression level [ ( 15.07 ± 0.83 ) %, ( 23.79 ± 3.99 ) %, (38.51 ± 1.58 ) %, (53.86 ±1.74)%;(15.43 ± 1.45)%,(36.11 ± 1.37)%, (56.86 ± 1.97)%, (76.09 ± 2.01)%; (15.20 ± 1.03)%,(35.25 ±1.09)%, (55.56 ± 2.17)%, (74.48 ± 2.85)% ] was respectively increased in a dose-dependent manner with the increased concentration of NaAsO2(all P < 0.05). The bel-2 mRNA expression level [ (443.00 ± 244.47), (156.79 ±53.18), (62.13 ± 13.66), (23.10 ± 6.44); (420.55 ± 110.77), (48.15 ± 10.02), (14.91 ± 6.53), (7.54 ± 2.62);(577.75 ± 123.22), (49.68 ± 10.11), (12.41 ± 1.28), (7.22 ± 1.89)], bcl-2 protein expression level[(47.20 ±3.77)%, (41.80 ± 2.94)%, (36.00 ± 2.36)%, (29.00 ± 2.91)%; (45.90 ± 4.15)%, (35.70 ± 2.77)%, (29.80 ±2.78)%, (24.80 ± 2.66)% ; (46.70 ± 3.47)%, (36.20 ± 2.90)%, (30.10 ± 3.21)%, (25.10 ± 2.28)% ] wasdecreased in a dose-dependent manner with the increased concentration of NaAsO2(all P < 0.05 ). In 3,9,15 mmol/L NaAsO2, apoptotic rate of p53 group, mRNA expression of bcl-2, protein expression of bcl-2 was higher than that ofnormal group and PC group, respectively (all P < 0.05), but mRNA expression of Bax, protein expression of Bax was respeetivelylower than that normal group and the PC group(P < 0.05 ). Conclusion p53 gene reduced the apoptosis induced by NaAsO2 in HELF, possibly by changing the apoptosis pathway.

Objective To report the experience of using rectus abdominis musculocutaneous flap after open-window thoracosto- my in managing refractory chronic pleural empyema.Methods From Nov.2004 to Mar.2006,intrathoracic transplantation of the musculocutaneous flap was performed successfully in 4 patients with empyema and fistula after upper lobectomy.The myocutaneous flap was designed in such a way that epigastrica vessels were incorporated.Results The rectus abdominis myocutaneous flap has provided sufficient bulk for tract obliteration.Over a mean follow-up period of 10 months,patients are free from recurrent infections and skin nacrosis.Conclusion Vaseularized museulocutaneous flap may help in healing refractory empyema.

Objective To explore the related protein which lead to rheumatoid arthritis (RA) and to find different proteins associated with active RA by comparing the expression levels of proteins in the peripheral blood mononuclear cells (PBMCs) of healthy individuals to patients with rheumatoid arthritis using a proteomics approach. Methods Samples of peripheral blood were collected from 9 patients diagnosed as active RA and 9 healthy individuals. PBMCs were isolated from blood using lymphoeytes separation medium. The total protein was extracted from the peripheral blood mononuclear cells. The total protein from either RA patients or normal controls was prepared by means of immobilized pH gradient based on two-dimensional gel eleetrophoresis. After Coomassie brilliant blue G250 staining, gel-image analysis was performed by using PDQuest.The differentially expressed proteins were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALD I-TOF-MS). Then APOA-I was validated by RT-PCR. Results 2-DE patterns of PBMCs from controls and RA patients were presented. The results showed that the average number of protein spots was 556 and 579 respectively, and the corresponding average matching rate was 89.4％ and 88.5％ respectively. Gel-image analysis revealed that there were 23 differential protein spots. Fourteen of total 18 differential protein spots were successfully identified by MALD I-TOF-MS, of which 8 proteins were upregulated such as actin beta, fibrinogen beta chain, ApoA-I ; and 6 proteins such as peroxiredoxin-2, glu-tathione S-transferase omega 1 were down-regulated when compared with controls. The result of ApoA-I by RT-PCR was consistent with the proteomics results. Conclusion Some differentially expressed proteins are observed in the PBMCs of patients with rheumatoid arthritis, which may play a potential role in the pathogenesis of RA.

Antigens, CD ; metabolism ; Antigens, CD34 ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Arteriovenous Malformations ; complications ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Magnetic Resonance Imaging ; Middle Aged ; Multiple Sclerosis ; Spinal Cord Diseases ; complications ; metabolism ; pathology

This study was undertaken to investigate the correlation of the enhancement degree on contrast-enhanced ultrasound (CEUS) with the histopathology of carotid plaques and the serum high sensitive C-reactive protein (hs-CRP) levels in patients undergoing carotid endarterectomy (CEA).Carotid CEUS was performed preoperatively in 115 patients who would undergo CEA,and the enhancement degree of the carotid plaques was evaluated by both the visual semiquantitative analysis and the quantitative time-intensity curve analysis.Serum hs-CRP levels were detected using the particle-enhanced immunoturbidimetric assay also before the operation.Additionally,the carotid plaque samples were subjected to histopathological examination postoperatively.The density of neovessels and the number of macrophages in the plaques were assessed by immunohistochemistry.The results showed that among the 115 patients,grade 0 plaque contrast enhancement was noted in 35 patients,grade 1 in 48 patients and grade 2 in 32 patients.The degree of plaque enhancement,the density ofneovessels,the number of macrophages,and the hs-CRP levels were highest in the grade 2 patients.Correlation analysis showed that the enhancement degree of the carotid plaques was closely related to the immunohistochemical parameters of the plaques and the serum hs-CRP levels.It was suggested that the carotid plaque enhancement on CEUS can be used to evaluate the vulnerability of carotid plaques.