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.

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 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.

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 effect of intrafrontal-orbital distraction osteogenesis in the treatment for school-aged children with severe proptosis in Crouzon syndrome.Methods:We retrospectively analyzed the clinical data of school-aged children with Crouzon syndrome treated with intrafrontal orbital distraction osteotomy at the Children’s Hospital of Nanjing Medical University from January 2016 to February 2021, and counted and analyzed the clinical results, complications (cerebrospinal fluid leakage, infection, etc.), and surgical safety.Results:A total of 12 children, 9 males and 3 females, aged 6-12.1 years, with an average age of 7.1 years, were included; 4 were cranial, 6 were craniofacial, and 2 were other types. All were shown with obvious proptosis. The surgical procedure was successful. Follow-up ranged from 3 to 20 months, with a mean of 14 months. The postoperative distraction distance was (19.1±3.3) mm, ranging from 15.2 to 25.6 mm. And the mean cranial index (CI) before and after surgery were 97.5% and 80.0%, respectively, with good craniofacial appearance and better improvement of proptosis and posterior cranial flattening. There were no other complications but for one child with infection at the external site of the distraction rod.Conclusions:The effect of frontal-orbital distraction osteotomy in the treatment of Crouzon syndrome with severe proptosis in school-aged children was positive, with significant improvement in proptosis and low incidence of postoperative cerebrospinal fluid leakage and infection. This procedure is a good choice for older children with severe proptosis.

Objective:To investigate the application and effect of frontal-orbital reconstruction and distraction osteogenesis in the treatment of Crouzon syndrome using three-dimensiond reconstruction mold (3D-RM).Methods:Retrospective analysis was performed on the clinical data of children with Crouzon syndrome who were treated in the Department of Burning and Plastic Surgery, Children’s Hospital of Nanjing Medical University from January 2018 to December 2021. These children were divided into two groups. The study group performed fronto-orbital reconstruction combined with distraction osteogenesis using 3D-RM, while the control group performed the surgery without using 3D-RM. Intraoperative blood loss, operation time, duration of hospital stay, complication rate, cranial index, frontal and orbital retraction rate and postoperative frontal and orbital contour were compared between the two groups. The independent sample t-test was used to compare the differences in intraoperative blood loss, operation duration, hospital stay, postoperative cranial index and fronto-orbital retraction rate between the two groups. The Chi-square test was used to compare the differences in the incidence of postoperative complications between the two groups. Results:The study group included 10 children, including 6 males and 4 females, ranging in age from 9 to 37 months, with an average age of 19 months. In the control group, 8 children were included, including 5 males and 3 females, ranging in age from 8 to 46 months, with an average age of 25 months. The study group was followed up for 12 to 36 months, with an average of 18 months, and the head shape was better improved. The control group was followed up for 12 to 30 months, with an average of 18 months. No serious complications occurred and the head deformity was improved. There were no significant differences in intraoperative blood loss, operation duration, hospital stay, cranial index, fronto-orbital retraction rate and complication rate between the two groups (all P>0.05). The forehead shape of the study group was more symmetrical than that of the control group, the difference was statistically significant ( P<0.05). Conclusion:The effect of frontal orbital reconstruction and distraction osteogenesis using 3D-RM in the treatment of Crouzon syndrome is positive, the head shape and orbit are significantly improved. There were no serious complications after operation. It can play a good auxiliary role in the operation.