Objective To construct the pCDNA3.1-ING4 eukaryotic expression vector and investigate its effect on the proliferation of melanoma cell line M14. Methods The targeted cDNA fragment encoding ING4 was cloned by reverse transcription-PCR with normal gastric mucosa from patients with gastric ulcer, and subcloned into eukaryotie expression vector pcDNA3.1. PCR and DNA sequencing were performed to identify the eukaryotic expression vector pCDNA3.1-ING4, which was then transfected into M14 cells with Lipofectamine 2000 reagent. The expression of ING4 was detected in untransfected, pCDNA3. 1-ING4-transfected and pcDNA3.1-transfected M14 cells by Western blot and immunocyto-chemistry, and cell proliferation by MTT assay. Results A fragment of expected size (750 bp) was amplified by PCR analysis, and DNA sequencing confirmed the correctness of the recombinant plasmid. As shown by immunocytochemistry, the percentage of cells positive for ING4 protein was significantly higher in pCD NA3. 1-ING4-transfected MI4 cells than in non-transfected M14 cells and pcDNA3.1-transfected M14 cells (71.80%±9.88% vs 4.20%±3.35%, P < 0.01). Western blot also revealed an increased expression of ING4 in pCDNA3.1-ING4-transfected cells. Decreased cell viability was observed in ING4-transfected cells compared with nontransfected cells and pCDNA3.1-transfected cells (both P < 0.01). Conclusions The pCDNA3. 1-ING4 eukaryotic expression vector has been constructed successfully, and M14 cells transfected by this recombinant plasmid could effectively express ING4 protein, which may inhibit the cell proliferation of M14 cells.

BACKGROUND:Tibial plateau fractures involving the posterolateral articular surface present a great challenge due to its complex anatomy.The posterolateral corner complex and the proximity of the common peroneal nerve restrict both the exposure of the joint surface and the ability to distract across the joint using a varus force.OBJECTIVE:To analyze the anatomy,morphology and classifications of the posterolateral fractures of the tibial plateau,and compare different surgical approaches and fixation materials.METHODS:A computer-based research of PubMed and CNKI databases was performed for the relative literatures published from 1990 to 2016 using the keywords of tibial plateau,posterolateral fragment,fixation in English and Chinese,respectively.The anatomy,morphology and typing of posterolateral fractures of the tibial plateau were analyzed and different surgical approaches and fixation materials were compared.RESULTS AND CONCLUSION:Posterolateral fractures of the tibial plateau should be treated in accordance with the intra-articular fracture principle,such as repairing joint surface to normal morphology,restoring the lower limb alignment,and maintaining joint mobility painlessly.Fractures are clarified according to Schatzker,AO,and three-column classification systems,but still need to be improved.To choose a proper surgical approach mainly depends on the fracture pattern.Biomechanics tests suggest that posterolateral locked plate exhibits excellent biomechanics characters,and can avoid the reduction loss caused by the shear force of fracture fragments.

Objective To investigate the current situation and related risk factors of healthcare-associated infection (HAI),so as to provide evidence for making prevention and control measures of HAI.Methods On November 26, 2014,a combination method of bedside visiting and medical record reviewing was adopted to survey HAI status, pathogen examination,and antimicrobial application in all hospitalized patients in a tertiary first class military hospi-tal.Results A total of 1 657 hospitalized patients were investigated,66 patients developed 71 times of HAI,HAI rate and HAI case rate were 3.98% and 4.28% respectively.The top 4 departments with HAI prevalence rates were departments of neurosurgery (24.49%),hematology(19.05%),cadre ward(13.73%),and burn surgery (10.91 %).The top 5 HAI sites were lower respiratory tract (40.85%),urinary tract(23.94%),upper respirato-ry tract(12.68%),surgical site(9.86%),and gastrointestinal tract(5.63%).Of 66 cases of HAI,39 (59.09%) patients sent specimens for culture,a total of 48 pathogens were cultured,the major isolated bacteria was Esche-richia coli (n = 10,20.84%),followed by Staphylococcus aureus ,Klebsiella pneumoniae ,and Pseudomonas aeruginosa ,each was 7 (14.58 %)respectively.The usage rate of antimicrobial agents was 34.40%,specimen de-tection rate in patients receiving therapeutic and therapeutic+prophylactic antimicrobial agents was 59.28%.Risk factors for HAI were age <2 years old or >60 years,with respirator,tracheotomy,urinary tract catheterization, arteriovenous catheterization,hemodialysis,and surgery,difference was significant(all P <0.05).Conclusion Mo-nitoring on key departments and key sites of HAI should be strengthened,antimicrobial agents should be used rationally based on pathogenic detection results,specimen pathogenic detection rate should be improved,and effective prevention and control measures needs to be taken according to the risk factors of HAI.

Objective To investigate the cachexia morbidity among hospitalized patients with digestive system cancer and evaluate its impact on clinical outcomes.Method By analyzing the clinical data of 5 118 hospitalized patients with digestive system cancer in Zhongshan Hospital,Fudan University from January 2012 to December 2013,we investigated the cachexia morbidity and compared the clinical outcome between cachectic patients and noncachectic patients.Results The overall cachexia morbidity of hospitalized patients with digestive system cancer was 15.7％ (803/5 118).The highest cachexia morbidity was 34.0％ (89/173),found in patients with pancreatic cancer.In cachectic group and non-cachectic group,the overall completion rate of radical resection was 67.1％ (539/803) and 74.5％ (3 214/4 315),respectively (P =0.000).Compared to the non-cachectic group,the cachetic group had significantly longer postoperative hospital days [(11.5 ±6.2) d vs (9.4 ±4.9) d,P =0.003],slower postoperative recovery of bowel function [(3.4 ±0.9) d vs (3.2 ±0.8) d,P =0.013],longer postoperative time to intake semifluid [(4.4 ± 1.5) d vs (3.9 ± 1.3) d,P =0.002],and more postoperative complications in 28 days after surgery [8.9％ (48/539) vs 5.8％ (186/3 214),P=0.006].After surgery,131 patients in the cachectic group were transferred to the ICU,and 646 patients in non-cachectic group transferred to the ICU (24.3％ vs 20.0％,P=0.026).Compared to the non-cachecic group,the reoperation rate [3.2％ (17/539) vs 1.5％ (48/3214)],ventilator support rate [8.0％ (43/539) vs 5.7％ (184/3 214)],and mortality [2.4％ (13/539) vs 1.1％ (35/3 214)] of the cachectic group were all significantly higher (P =0.006,0.042,0.011).Conclusions Cachexia is common in hospitalized patients with digestive system cancer,especially in patients with pancreatic cancer.Cachexia has negative impact on the clinical outcomes.

Objective To provide therapeutic strategy by investigating the anxiety and depression status as well as the correlated impact factors in patients with chronic prostatitis. Methods The self-rating anxiety scale (SAS)and self-rating depression scale (SDS)were employed to evaluate psychological response of 360 patients with chronic prostatitis. Results Both the average scores of SAS and SDS in patients with chronic prostatitis were higher than normal levels of Chinese population. The impact factors for anxiety included income status, educational level and disease course. Conclusions The mental health status of chronic prostatitis patients is poor and certain factors affect the occurrence of anxiety and depression. Therefore, it is necessary to treat the patients with supportive psychotherapeutic intervention and with combined approaches.

Objective To explore the diagnostic value of radial ultrasound (EBUS) combined with virtual navigation (VBN) in peripheral pulmonary nodules.Methods Two hundreds and forty cases of peripheral pulmonary nodules(0.8 cm≤diameter≤3 cm) in the respiratory department of our hospital from July 2014 to July 2015 were included and according to the different guide de vices,which were divided into the radial ultrasound combined with virtual navigation group (EBUS+ VBN group),radial ultrasound group (EBUS group),virtual navigation group (VBN group) and control group.The diagnostic rates were compared among the four groups and among different sizes of lesion.The time of lesion location and operating time were also compared between the EBUS+ VBN group and EBUS group.Results Among 240 cases,the diagnostic rate in the EBUS+VBN group was highest(81.67％),and the diagnostic rates had statistically significant difference among the four groups (x2=19.344,P=0.00);the diagnostic rates of lesions less than 2 cm in diameter were lower than that of lesions ＞2 cm in the EBUS+-VBN group and EBUS group,but without statistically significant difference (x2 =2.04,3.40,P =0.15,0.07);the locating lesions time and operating times in the EBUS+ VBN group were shorter than those in the EBUS group,but the difference between them was not statistically significant (P=0.03,0.04).Conclusion EBUS combined with VBN could improve the diagnostic rate of peripheral pulmonary nodules and shorten the time of lesion location and operating time.

Objective To characterize the clinical manifestations and to evaluate the value of the electrophysio-logical examination in patients with peripheral nerves injuries caused by neurobrucellosis. Methods Electrophysiology examination was conducted in 32 patients with peripheral nerve injuries caused by neurobrucellosis and 32 normal controls who had equivalent age and gender. The results were further statistically analyzed. Results There were sig-nificant difference between patients and healthy control group in distal motor latency (DML), compound motor active potentials (CMAP) amplitude, motor nerve conduction velocity (MCV), sensory nerve action potential latency(SL),senso-ry nerve action potential (SNAP) amplitude and sensory nerve conduction velocity (SCV) ( P﹤0.05). Electrophysiology examination revealed peripheral nerve damage of limbs involving, both sensory nerve and motor nerves, of which sen-sory and motor nerve injuries accounted for 55.47% and 16.80%, respectively. Median and sural nerve injuries were most frequently affected in upper (64) and lower limbs (16). Motor and sensory nerve conduction velocity was delayed in 43 nerves (16.80%) of 256 examined motor nerves and in 142 nerves (55.47%) of 256 examined sensory nerves. The damage of the sensory nerves was more severe than those of the motor nerves and damage of nerves in the upper limbs was more severe than those in the lower limbs. Conclusion Electrophysiologoical examination provides objective basis for the diagnosis of peripheral nerve injuries in neurobrucellosis.

Objective To explore the clinical effect of periarticular knee fractures combined with anterolateral ligament (ALL) avulsion fracture.Methods From June 2014 to November 2015,13 patients with ALL avulsion fracture in periarticular knee fractures were fixed by the star plate,cannulated or screws suture anchor,including 9 male and 3 female with the mean age of 39.5 years (range from 28 to 51 years).6 patients were diagnosed distal femoral fracture,4 tibial plateau fracture,and 3 tibial intercondylar eminence avulsion fracture which all combined with ALL avulsion fracture.Postoperatively,all patients were treated with the adjustable knee brace for 3 months.Results All patients were followed up for 4-20 months,the mean follow-up time was 13.5 months.The average surgical time was 130 min (range from 90 to 210 min).In all 13 patients,ALL was found and identified.Bone union was obtained in all patients,and the bone union time was 11.4 weeks (10-12 weeks).7 cases were fixed with star plate,5 cases were fixed with hollow screw,and 1 case was fixed with anchor.All follow-up patients underwent anteroposterior and lateral X-rays.Pain,swelling and functional recovery of the knee joint were observed.Patients were assessed postoperatively with International Knee Documentation Committee (IKDC) score,Lysholm score,and Tegner activity scale to evaluate the clinical effect.At the last follow-up,all patients had full range of motion of the knee joint which from 0° to approximate 120°.The Lysholm,objective IKDC were 86.5 and 84.6 respectively.Among the 13 patients evaluated,9 patients were graded A,and 4 patients were graded B by subjective IKDC.The Tegner activity scale at the last follow-up was 6.7.Incision of poor healing in 1 case,and got healed after 32 d cleaning dressing.Conclusion This study confirmed the presence of the ALL.The fixation of ALL avulsion fracture in periarticular knee fractures can be an effective procedure at a minimum follow-up of 13.5 months.

BACKGROUND:Fetal bovine serum as nutritional support is often used in the traditional cel culture. Consequently, a host of potential problems such as the spread of disease and immunological reactions exist. To find a suitable fetal bovine serum substitute and to establish a culture system of human bone marrow stromal stem cels in vitro which has been standardized, safe and efficient has just started. OBJECTIVE:To investigate the effects of different serums on proliferation of bone marrow stromal stem celsin vitro. METHODS:Bone marrow stromal stem cels were obtained from adult bone marrow, which were cultured in DMEM containing 10% AB serum, 10% autologous serum, or 10% fetal bovine serum. Cels at passage 3 were used in this study. RESULTS AND CONCLUSION:The cel confluence in the AB serum group was earlier than that in the fetal bovine serum group and autologous serum group. Human bone marrow stromal stem cels maintained the phenotypes of bone marrow stem cels in three serums detected by flow cytometry. AB serum group showed the highest fluorescence intensity and the most efficiency of cel proliferation which examined by the AlamarBlue assay. Apoptosis rate was < 5% in al the three groups, and cels grew wel in these serums. Alkaline phosphatase, calcium nodules and oil red O staining showed that the cels maintained the osteogenesis and adipogenesis capacity in the three groups. AB serum was found to have a better effect on proliferation capability of cels than fetal bovine serum and autologous serum. Taken together, AB serum is expected to be a substitute of fetal bovine serum to build anin vitro culture system of adult bone marrow stromal stem cels that accord with the clinical requirements of bone tissue engineering.

AIM:The objective of this study was to observe the effects of AP-1 element on endothelin-1(EDN1)gene transcription in homocysteine(Hcy)-induced human umbilical vein endothelial cells(HUVECs).METHODS:The redox level in Hcy-induced HUVECs was determined by using the fluorescent product DCF.The influences of Hcy on expressions of EDN1 mRNA and protein of c-jun/AP-1 in HUVECs were measured by the methods of RT-PCR and Western blotting,respectively.The EDN1 secretion level of HUVECs induced by Hcy was determined by enzymatic immunoassay.The transient transfection assay was performed and luciferase activity of transcriptional factor AP-1 reporter gene induced by Hcy was detected.RESULTS:The Hcy significantly increased EDN1 secretion,EDN1 mRNA expression and oxidative stress in HUVECs.Hcy remarkably induced the expression of pGL3-EDN1-AP-1(115/+135)luciferase reporter gene plasmid.However,basic expression of mutation of pGL3-EDN1-AP-1(-115/+135)luciferase reporter gene plasmid was downregulated markedly in HUVECs induced by Hcy.CONCLUSION:Redox-active and release of ROS are changed by Hcy.Activated AP-1 element plays an important role in EDN1gene transcription in HUVECs induced by Hcy.