Objective To investigate the risk factors of incision infection in elderly patients of general surgery.Methods The clinical data of 306 elderly patients of general surgery were analyzed retrospectively.The risk factors of incision infection were analyzed.Results The rate of incision infection was 10.5％ (32/306).The rate of incision infection in chronic disease patients was higher than that in non-chronic disease patients [12.9％(27/210) vs.5.2％(5/96)],the rate of incision infection in summer was significantly higher than that in spring,autumn and winter[16.3％(15/92) vs.8.4％(7/83),8.5％(6/71) and 6.7％ (4/60)],the rate of incision infection in operative time longer than 120 min patients was higher than that in operative time less than 120 min patients [15.0％ (18/120) vs.7.5％ (14/186)],the rate of incision infection in emergency surgery patients was higher than that in non-emergency surgery patients [12.3％ (20/162) vs.8.3％(12/144)],there had significantly difference (P＜ 0.05).Conclusion Chronic disease,summer,operative time longer than 120 min,emergency surgery in elderly patients are the mainly risk factors of incision infection in general surgery.

Objective To investigate the effect of the Ras-related tumor suppressor gene aplasia Ras homolog member Ⅰ (ARHI) on angiogenesis in hepatocellular carcinoma (HCC).Methods We generated stable cell lines overexpressing ARHI in Hep3B cells,which lack endogenous ARHI.Cell proliferation was assessed by the MTT assay.The effects of ARHI overexpression on tumor growth and angiogenesis were assessed.Because of the key role of mammalian target of rapamycin (mTOR)signaling in HCC progression,we also tested whether ARHI overexpression affected the mTOR pathway.Results Ectopic expression of ARHI significantly diminished cell proliferation in Hep3B cells (P＜0.01).ARHI overexpression significantly retarded Hep3B xenograft growth by 76.4 ％ in vivo,and caused a marked reduction in tumor angiogenesis assessed by CD31-stained microvessel count.Western blot analysis of the xenografts showed that ARHI overexpression substantially reduced the phosphorylation of two mTOR substrates,S6K1 and 4E-BP1,indicative of an inactivation of the mTOR pathway.Accompanying with the mTOR inactivation,the angiogenic factors,hypoxia-inducible factor 1 alpha and vascular endothelial growth factor,were significantly downregulated.Conclusion These data highlighted an important role for ARHI in controlling HCC growth and angiogenesis,therefore offering a possible therapeutic strategy against this malignancy.

AIM:To explore the protective effect of fasudil hydrochloride against cisplatin (CP)-induced renal tubular epithelial cell apoptosis via Akt activation and PTEN inhibition .METHODS:Healthy male Sprague-Dawley ( SD) rats were randomly divided into control group , CP group and CP+fasudil group .All animals were sacrificed 96 h after in-jection of 0.9%saline or CP .Blood samples and kidney tissues were collected to evaluate levels of blood urea nitrogen (BUN), serum creatinine (sCr) and morphological alteration of the kidneys , respectively.The apoptosis of renal tubular epithelium cells was detected by TUNEL.Protein levels of Rho-associated protein kinase 1 (ROCK1), PTEN and Akt were measured by Western blotting and immunohistochemistry .The protein level of p-Akt was analyzed by Western blotting . RESULTS:Compared with control group , the sCr and BUN levels , the expression of ROCK 1 and PTEN and TUNEL-posi-tive cells were increased , while the level of p-Akt was decreased in CP group and CP +fasudil group .The histological structure of the kidneys observed by PAS staining was developed marked structural damage in CP group (P<0.05).Com-pared with CP group, sCr level, the expression of ROCK1 and PTEN and TUNEL-positive cells were decreased, while the level of p-Akt was increased in CP+fasudil group (P<0.05).Very little structural damage was detected in fasudil-treated groups .CONCLUSION:Fasudil hydrochloride has a protective effect on CP-induced renal tubular epithelial cell apoptosis via Akt activation and PTEN inhibition 1.

Objective To evaluate the feasibility and clinical efficacy of amputation in situ secondary braches of splenic pedicle for laparoscopic splenectomy(LS)in patients of idiopathic thrombocytopenic purpura(ITP).Methods LS was performed in 41 ITP patients hospitalized in our department from January 2007 to November 2008 by dissecting secondary braches of splenic pedicle.The serosa on the spleen pedicle was opened by using a harmonic scalpel.Then secondary structures of the splenic arteries and veins,one by one,were disconnected from downward to upward,and then double ligated with the Hem-O-lok clips.The ligated section was cut off with the harmonic scalpe1,and then the spleen was resected.Results LS was successfully completed with no conversion to open surgery in all the 41 cases.The operative time was 75 to 180 min,and the estimated intra-operative blood loss was 50 to 800 ml.There were 3 cases of vice spleen and the vice spleen removed.Two cases were converted to traditional LS with Endo-GIA.The period of hospitalization was 4 to 9 d after operation.There was no severe postoperative complication.Platelet counts began to increase significantly after operation,reached to the peak in 5 to 7 d after operation.During the follow-up of 3 to 20 months to all 41 ITP patients,the total effective rate was 85.4%(35/41,including 22 cases of complete response and 13 cases of partial response).Conclusion LS with amputation in situ secondary braches of splenic pedicle is a safe,effective,minimally invasive and low complication in treatment of ITP.It is worth be recommended because of its lower costs in comparison with common LS.

[ ABSTRACT] AIM:To investigate the protective effect of naringin ( Nar) on the injury of human umbilical vein endothelial cells ( HUVECs) induced by 33 mmol/L high glucose ( HG) and to explore its possible mechanisms.METH-ODS:The injury model was established by treating HUVECs with HG medium for the indicated time (6, 12, 24, 48 and 72 h) , and then the levels of NO, eNOS and p-eNOS were detected, respectively.The effects of Nar on high glucose-in-duced endothelial cell injury were observed.HUVECs were treated with Nar at concentrations of 5, 10, 25, 50 and 100 mg/L for 6 h, 12 h, 24 h, 36 h and 48 h.The levels of NO in the supernatants were measured.The effects of Nar on HG-injured HUVECs were explored by treating the cells with 10 μmol/L of LY294002, a PI3K inhibitor, or 0.5 μmol/L of AKT inhibitorⅣ, an AKT inhibitor, and then the levels of NO, PI3K, AKT, eNOS and their phosphorylated proteins were determined by Western blot.RESULTS:Nar at concentration of 50 mg/L significantly attenuated the injury of endothelial cells induced by high glucose ( P<0.01) , and the protective effects of Nar were abolished by pretreating with the inhibitor of PI3K or AKT (P<0.01).CONCLUSION: Nar protects endothelial cells against the injury induced by high glucose through PI3K/AKT/eNOS pathway.

Objective: To explore the effects of Bmi1 on proliferation of mouse cranial suture mesenchymal cells. Methods: Primary posterior frontal and sagittal suture derived cells were isolated from the 2-5 d old C57BL/6 suckling mice (n=6) of the same brood and cultured. Flow cytometry and multilineage differentiation assay were performed to identify the mesenchymal stem cells (MSCs) characteristics of the 2 kinds of cranial suture-derived cells. The mRNA expression of stem cell related genes, Bmi1, Twist1, Gli1 and Axin2 were detected by real-time quantitative polymerase chain reaction (RT-PCR). Then, the proliferation and downstream protein expression were analyzed after down-regulation of Bmi1 in the sagittal suture derived MSCs by transfecting Bmi1 siRNA. The t test was used to compare the mean between two groups. Statistical significance was set at P<0.05. Results: The mouse cranial suture derived cells were successfully cultured in vitro. These cells expressed typical MSCs markers, CD44, CD90, CD73, except for CD34. These cells had osteogenic, adipogenic and chondrogenic differentiation potency. RT-PCR results showed that the mRNA expressions of Bmi1 (0.006 30±0.000 58 vs 0.002 60±0.000 34, t=5.430, P=0.005 6), Twist1(0.000 31±0.000 04 vs 0.000 15±0.000 02, t=3.343, P=0.028 8), Axin2(0.000 33±0.000 03 vs 0.000 17±0.000 05, t=3.067, P=0.037 4) and Gli1 (0.001 10±0.000 13 vs 0.000 60±0.000 33, t=3.956, P=0.016 7) were significantly decreased in the posterior frontal suture MSCs compared with those in sagittal suture derived cells. Among them, Bmi1 has the largest decline. After down-regulation of Bmi1 in sagittal suture MSCs, the protein expression level of Ink4a was significantly up-regulated compared with the control group, and the cell proliferation ability was significantly decreased. Conclusions Inhibition of Bmi1 expression can up-regulate the expression of Ink4a protein and decrease the proliferation ability of suture MSCs, which may lead to craniosynostosis.

Background and Objectives: Cranial sutures play a critical role in adjustment of skull development and brain growth. Premature fusion of cranial sutures leads to craniosynostosis. The aim of the current study was to culture and characterize human cranial suture mesenchymal cells in vitro. Methods: The residual skull tissues, containing synostosed or contralateral suture from three boys with right coronal suture synostosis, were used to isolate the suture mesenchymal cells. Then, flow cytometry and multilineage differentiation were performed to identify the typical mesenchymal stem cell (MSC) properties. Finally, we used quantitative real-time polymerase chain reaction (RT-PCR) to detect the mRNA expression of osteogenesis and stemness related genes. Results: After 3 to 5 days in culture, the cells migrated from the tissue explants and proliferated parallelly or spirally. These cells expressed typical MSC markers, CD73, CD90, CD105, and could give rises to osteocytes, adipocytes and chondrocytes. RT-PCR showed relatively higher levels of Runx2, osteocalcin and FGF2 in the fused suture MSCs than in the normal cells. However, BMP3, the only protein of BMP family that inhibits osteogenesis, reduced in synostosed suture derived cells. The expression of effector genes remaining cell stemness, including Bmi1, Gli1 and Axin2, decreased in the cells migrated from the affected cranial sutures. Conclusions The MSCs from prematurely occlusive sutures overexpressed osteogenic related genes and down-regulated stemness-related genes, which may further accelerate the osteogenic differentiation and suppress the self-renewal of stem cells leading to craniosynostosis.

Objective:To construct the pET30a-EgG1Y162-2 prokaryotic expression plasmid and induce the expression of EgG1Y162-2 protein, so as to provide a research basis for development of Echinococcus granulosus vaccine. Methods:Using Echinococcus granulosus cDNA as a template, the target gene of EgG1Y162-2 was synthesized by PCR, and after digestion with restriction enzymes EcoRⅠ and Hind Ⅲ, it was connected to the prokaryotic expression vector pET30a to construct the recombinant plasmid pET30a-EgG1Y162-2. The recombinant plasmid was transformed into competent cell BL21 (DE3) and induced by isopropyl β-D-thiogalactoside (IPTG) to express a large number of proteins. The recombinant protein was purified by affinity chromatography. The purification level was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the expression product was identified by Western blotting. Results:The recombinant plasmid pET30a-EgG1Y162-2 was successfully constructed. After inducting expression, the bacterial supernatant and the eluate were both at a relative molecular weight of about 15 × 10 3, and the protein antigen component eluted with 200 mmol/L imidazole was relatively pure. Western blotting results showed that the purified recombinant protein EgG1Y162-2 with His tag could be recognized by His monoclonal antibody. Conclusion:The pET30a-EgG1Y162-2 prokaryotic expression plasmid of Echinococcus granulosus is successfully constructed, and the recombinant protein of EgG1Y162-2 is induced to express, laying a foundation for further study on anti- Echinococcus granulosus vaccine.

Objective:To investigate the treatment of craniosynostosis in small infants with posterior vault distraction osteogenesis and the reasonable treatment for lambdoid sutures during operation.Methods:Seven cases with syndromic craniosynostosis who were admitted to our department from January 2017 to August 2019 were osteotomized by piezosurgery supraoccipital protuberance osteotomy in the posterior cranium, and two 2 cm distractors were placed in the lateral cranium. In 4 of 7 cases bilateral lambdiod sutures were fixed with titanium plate, while 3 of 7 were not fixed. Distraction was started from 5 days postoperatively, once a day on each side with a rate of 0.4-0.8 mm/d on each side for 10-15 days. The distractors were removed 6 months postoperatively. The titanium plates fixation in 4 cases were also removed at the same time.Results:The patients were followed up for 6 to 14 months, with an average of 12 months. The skull cavity and posterior cranial shape were observed. The posterior vault was distracted completely with lambdoid suture fixation in 4 cases. In the other 3 cases, the posterior skull was flattened. All skull deformities were significantly improved without death, skull necrosis, intracranial infection and other serious complications. The cranial index of both groups of lambdoid suture fixed and unfixed were reduced after the operation, and the fixed group was reduced by an average of 20.5%±5.8%, higher than the unfixed group(17.5%±5.1%), but the difference between the two groups was not statistically significant( P>0.05). The occipitofrontal circumference was increased after the operation in the two groups. The fixed group was increased by (2.7±0.4)cm on average, which was significantly higher than the unfixed group(1.7±0.1) cm. And the difference was statistically significant( t=3.789, P=0.013). Conclusions:Posterior vault distraction osteogenesis is a good method to treat the syndromic craniosynostosis. But bilateral lambdoidsuture needs to be fixed in infants. Titanium plate fixation could make the posterior cranium into a whole cranial plate for posterior vault distraction osteogenesis. And the posterior vault was obviously flat without lambdoid sutures fixation. Flatted occipital contour was improved significantly and the result of the fixed group was superior to the unfixed one. Bilateral fixation of lambdoid sutures is suitable for the infants, especially for severe syndromic craniosynostosis.

Objective:To investigate clinical effects of the three-dimensional(3D) digital technique-assisted distraction osteogenesis in newborns with Pierre Robin sequence(PRS).Methods:The data of neonatal Pierre Robin sequence treated with 3D digital technique-assisted distraction osteogenesis in Children’s Hospital Affiliated to Nanjing Medical University from April 2017 to April 2020 was retrospectively analyzed. 3D digital technique was used to assist the pre-operative design and osteotomy navigation template was made with 3D printing technique to guide in the mandibular osteotomy. CT scans were obtained 3 months postoperatively to follow up the growth of mandible. The rates of recurrence, titanium nail falling, infection, secondary operation and nerve injury were statistically analyzed. The counting data were analyzed using chi-square test. The independent influencing factors of postoperative complications were analyzed by multiple logistic regression, and the difference was statistically significant when P<0.05. Results:Three hundred and forty-five patients were included in the 3D digital technique group consisting of 215 males and 130 females with the ages ranged 3-28 days and the weights 1.8-3.2 kg. Surgical results were satisfied and very close to the preoperative design. Craniofacial CT results showed normal growth of mandible 3 months postoperatively. Three hundred and seventy-seven patients were included in the controlled group consisting of 230 males and 147 females with the ages ranged 6-28 days and the weights 1.6-3.6 kg. The recurrence rate, extender titanium nail off rate, reoperation rate and risk of nerve damage in the 3D digital technique group were 0.29%(1/345), 0.29%(1/345), 0.58% (2/345)and 0.29%(1/345), respectively, while in the control group, the numbers were 1.86%(7/377), 2.92%(11/377), 2.65%(10/377), 2.92% (11/377), respectively, which were significant lower than those in the 3D group. The infection rates were 7.83%(27/345) in the 3D digital technique group and 7.69%(29/377) in the control group, and there was no significant difference between the two groups. The results of multi-logistic regression analysis suggested that the 3D digital technique group could be an independent factor to reduce postoperative complications ( P=0.001). Conclusions:In the newborns with Pierre Robin sequence, 3D digital technique-assisted distraction osteogenesiscan significantly improve the accuracy and success rate of the operation and reduce the complication rate as well.