Objective Within human hepatoma cell lines,we aimed to investigate the effects of the down-regulation by RNAi on different fragments of osteopontin (OPN) in order to discover more effective and accurate sites for OPN.Methods Specific small interfering RNA of OPN (OPNi-1) were synthesized and transfected into human hepatoma cell line (HEP-G2).Fluorescent quantitative PCR and immunohistochemical methods were used to test*the OPN expression levels of mRNA and protein before and after RNAi.Results After transfection,the △CT value of the A fragment was greater than B and C fragments of OPN mRNA in HEP-G2.Before RNAi was added to HEP-G2 cells,the three fragments A,B,C had OPN mRNA CT values of 8.31±1.58,8.78±1.49,8.25±1.51 respectively.Once the RNAi were added,the CT values were measured 48h after for the fragments A,B,and C which were 12.14±1.43,10.22±1.97,10.48±1.88 (P＜0.05) respectively.The immunohis tochemical values of A,B,C were down from 6.44±1.67,5.43±2.05,5.45±2.52 to 2.84±1.52,4.43± 1.65,3.95± 1.43 respectively after interference.Conclusions RNAi can inhibit the expression of OPN gene selectively.siRNA targets different segments of OPN,which may have more effects on invasion and metastasis of liver cancer for a more important significance in science and health economics.

BACKGROUND: Subsequent to liver transplantation, pulmonary infection and new drug resistant strain frequently appear due to complex pathophysiological changes and abuse of antibiotics.OBJECTIVE: To collect the clinical data of 6 cases who underwent liver transplantation, and analyze the prevention and treatment measures of pulmonary infection after liver transplantation.DESIGN, TIME AND SETTING: Retrospective case analysis, performed in the Department of Hepatobiliary Surgery,First People's Hospital of Guangzhou between January and December 2004.PARTICIPANTS: Six cases that underwent liver transplantation and received treatment in the First People's Hospital of Guangzhou were recruited in the present study. All 6 cases underwent selective homologous whole liver transplantation, 5cases underwent modified orthotopic piggyback liver transplantation, and only 1 underwent classical orthotopic liver transplantation.METHODS: The clinical data of the 6 cases were retrospectively analyzed. After transplantation, basic disinfection,isolation, infection-protection measures should be taken. In addition, etiological information was collected. Chest X-ray was underwent to know the lung at the early stage. Aseptic manipulating was strictly performed during sputum aspiration.Gastrointestinal decompression and non-obstructed drainage were maintained to prevent aspiration. Moreover, preventive anti-infective treatment against the bacteria, fungi, and viruses was performed.MAIN OUTCOME MEASURES: Pulmonary infection and curative effects subsequent to liver transplantation.RESULTS: Of the 6 cases, 5 presented with pulmonary infection, including 1 at 4-11 days after transplantation and 1 at 1day after transplantation. One case died of respiratory failure. The remaining 5 cases were cured after standardized anti-infection, respiratory therapy, sputum aspiration, and nutritional support.CONCLUSION: Application of effective antibiotics, aseptic operation in sputum aspiration, and unobstructed respiratory tract drainage are important measures for treating pulmonary infection subsequent to liver transplantation.

Objective To evaluate the role of mangled extremity severity score(MESS)in res-ervation and amputation of crush lower limbs in earthquake. Methods There were 122 patients with crush lower limb injuries,with MESS≥8 points in 34 patients who were primarily amputated,M ESS 5-7points in 19 who were principally preserved and MESS＜5 points in 69 who were preserved by means of debridement,external fixators,plast splints and vaeuum sealing drainage technique.Results All pa-tients were survived.with amputation rate of 29.5%. Conclusion MESS is an important reference for evaluation of reservation and amputation of crush limb injuries caused by earthquake.

Objective To explore the characteristics of proinsulin secretion in latent autoimmune diabetes in adults (LADA). Methods Fasting and 2 h sera in oral glucose tolerance test from 36 LADA patients, 37 type 2 diabetic patients and 43 healthy controls were collected to test glucose, proinsulin (PI) and C-peptide (CP) by radioimmune assay. Glutamie acid decarboxylase antibodies (GAD-Ab) were determined by radioligand assay.Results (1) Fasting proinsulin (FPI) and 2 h proinsulin (PPI) level in LADA patients were lower than those in type 2 diabetic patients (P<0.05), being both significantly inereasad compared with healthy controls (P<0.05 or P<0.01); The ratios of FPI/FCP and PPI/PCP (%) in LADA were beth significantly higer than those of type 2 diabetes mellitus and controls (P<0.05 or P<0.01); (2) LADA type-1 (GAD-Abe>0.3) patients showed lower PI levels(P<0.05 or P<0.01) and higher PI/CP ratio (all P<0.05) than LADA type-2 (0.05≤GAD-Ab<0.3); Meanwhile, there were no significant differences in above parameters between LADA type-2 and type 2 diabetes meUitus (P>0.05). (3) GAD-Ab index was negatively correlated with FPI and PPI in LADA group (r=-0.236 and-0.268, both P<0.05), and positively correlated with PPI/PCP (r=0.254, P=0.030).Meanwhile BMI was positively correlated with FPI, PPI and PI/CP in type 2 diabetes mellitus (all P<0.01). No factor entered the multiple regression analysis for predieting the hyperproinsulinemia and dispropriately elevated proinsulin levels in LADA patients. (4) According to the 99.5 th percentile of proinsulinemia in the healthy controls, which is defined as the cutoff point dispropriately elevated proinsulin levels, the proportion of subjects with fasting dispropriately elevated proinsulin levels (FPI/FCP) were 77.8%, 62.2% and 2.3% in LADA, type 2 diabetes meUitus and controls respectively, and PPI/PCP 83.3%, 51.4% and 2.3% respectively. Conclusion LADA patients, as well as type 2 diabetic patients, all showed hyperproinsulinemia and disproportionately elevated proiasulin levels that were one of characteristics of defective β-cell function. Moreover, disproportionately elevated nproinsulin level is more evident in LADA patients than that in type 2 diabetics and this may be related to humoral immunity.

Objective To systematically evaluate the role of laparoscopic living donor hepatectomy in living donor liver transplantation (LDLT).Methods A systematic literature search was conducted on Medline-Pubmed,Embase,Cochrane Library to find studies on laparoscopic living donor hepatectomy for LDLT.All extracted data were analyzed using the RevMan 5 software.Results Ten studies with a total of 1 059 participants were included in this analysis.Laparoscopic donor hepatecomy (LDH) was associated with significantly less intraoperative blood loss [SMD =-0.39,95％ CI (-0.73,-0.05),P ＜ 0.05],lower peak level of postoperative total bilirubin [SMD =-0.24,95％ CI (-0.47,-0.01),P ＜ 0.05]and longer operative time [SMD =0.50,95％ CI (0.04,0.96),P ＜0.05] when compared with those operated with open surgery.On subgroup analyses,hospitalization stay decreased in patients who underwent LDLT with grafts obtained by complete living donor hepatectomy (LDH) and left lateral sectionectomy (both P ＜ 0.05).LDH was comparable to open surgery in donor complication rates and in-hospital cost (P ＞ 0.05).There were no differences on the harvested liver graft size,ischemic time,recipient postoperative liver function and complications between the two groups (P ＞ 0.05).Conclusions Laparoscopic hepatectomy in living donor is a safe procedure for graft-harvesting,which improved the clinical outcomes of the donor,liver graft and recipient in LDLT.It has also the advantages of reduced blood loss,low peak levels of postoperative total bilirubin and short hospitalization stay.

Objective To study the correlation of cleft lip and palate transmembrane 1 like(CLPTM1L)expression and radiosensitivity of lung cancer cells.Methods Thiazolyl blue tetrazolium bromide(MTT) and cell colony formation assays were used to determine cell growth and survival.Western Blot assay was employed to measure protein expression.Results The results demonstrated a negative correlation between the CLPTM1L expression level and radiosensitivity of lung cancer cells.A lower radiosensitivity in lung cancer cells containing high level of CLPTM1L expression,and vice versa.Enforced expression of CLPTM1L resulted in a significant reduction of radiosensitivity in lung cancer cells irradiated with γ-rays.On the contrary,a marked elevation of radiosensitivity was observed in lung cancer cells transfected with CLPTM1L siRNA.Conclusions CLPTM1L may be a novel target gene in mediating radiosensitivity of lung cancer cells.

Objective To investigate the risk factors and treatment for infections after spinal internal fixation surgery. Methods The clinical data of 472 patients who underwent spinal internal fixation surgery from January 2012 to December 2012 was analyzed retrospectively, an average age of 50. 6 years (38~78 years). All cases were underwent posterior procedures. All infected patients received emergency opera-tion of wound debridement, drainage and sensitive antibiotic treatment. The mean follow-up time was 11 months (8~19 months). Risk fac-tors and treatment for infections were summarized and discussed. Results Of 472 patients,postoperative infections occurred in 9 cases with the infection rate of 1. 91%. The operation time,intraoperative blood and postoperative drainage was 100~325 min,200~1500 mL and 65~1350 mL,respectively,which were greater than the similar surgeries of same period. The initial signs of wound infection was observed at 10 d (6~16 d) after surgery. CRP,ESR and WBC were significantly increased in 4~7 d after surgery,and maintained at high level at least for 14 d. Bacterial culture results showed infection bacteria were mainly common skin flora. One infection recurred during followed-up and subse-quent treatment was successful. Conclusion Wound infection after internal fixation mainly occurred in the posterior procedure of spine, which were deep infection. The main clinical manifestation was the wound exudate and local deep tenderness,fever and wound surface swelling were relatively rare. Increased intraoperative bleeding,postoperative drainage volume,operation time were the risk factors,which lead to perio-perative malnutrition and subsequent infections. Debridement,drainage,and intravenously sensitive antibiotics could obtain an ideal outcome for most cases. It was not necessary to remove the internal fixation instrument and bone grafting.

Objective To explore the gender difference in Chinese version of autism spectrum rating scale (ASRS)-in terms of parents and teacher rating.Methods A total of 701 children (380 boys,321 girls;6 -11 years old)were recruited from one Primary School in Minhang District of Shanghai.The parents and teachers of the children completed the ASRS.While the data were imput into the database,the software automatically calculated T -standard score according to the formula for the raw score of screening scale,treatment scale,and Diagnostic and Statistical Manual of Mental Disorders -Ⅳ -Turning Reversion scale.In the end,the gender differences in Chinese version of ASRS for both parents and teacher ratings were analyzed.Results A total of 671 parents rating scale were retrieved (response rate was 95.7%),involving 368 boys (54.8%);685 for teacher rating scale (response rate was 97.7%), involving 381 boys (55.6%).No significant difference was observed in the distribution of gender(P >0.05).Minor influence of age was found among ASRS sub -scales for both parents and teacher ratings in boys and girls (parent rating scale r =-0.219 1 to -0.124 4,teacher rating scale r =-0.328 0 to -0.120 0).In parent and teacher ver-sion of ASRS,boys had higher T -standard scores in all subscales by 1 -3 and 2 -4 points than those in girls,respec-tively,and apart from abnormal behavior,adult socialization,behavioral rigidity,and sensory sensitivity there was no sig-nificant difference(all P >0.05)in parents rating scale while others had significant differences(all P <0.05),and all sub -scales had significant difference in teacher rating scale(all P <0.01).Conclusions The T -standard scores of both ASRS questionnaire has the gender differences in ASRS subscales,so it is necessary for researchers to understand the Chinese version of ASRS questionnaire from different angles.

Objective To investigate the prevalence of chronic kidney disease (CKD)and risk factors in the adult population of Tianshan district in Urumqi, Xinjiang. Methods A total of 2131 residents from 4 communities in Tianshan district of Urumqi city were randomly selected using a stratified, multistage sampling. All the residents were interviewed and tested for morning spot urine of albumin to creatinine ratio (ACR) (abnormal ≥ 30 mg/g), morning spot urine dipstick of hematuria ( abnormal ＞3 red blood cells/HP or greater) and pyuria ( abnormal＞ 5 white blood cells/HP) confirmed by microscopy. Renal function was determined with abbreviated MDRD equation [reduced estimated glomerular filtration rate (eGFR)＜60 ml ·min-1 ·(1.73 m2)-1]. The associations of kidney damage indicators with age, gender, hypertension, diabetes mellitus, income,education, cholesterol, triglyceride and smoking were examined. Results Eligible data of 2131 subjects were collected in the study. After the adjustment of age and gender component, the prevalence of albuminuria was found in 2.63％ (95％CI:1.78％-3.48％) of subjects, hematuria in 7.43％(95％CI:6.11％-8.75％) and reduced renal function in 1.72％(95％CI:1.08％-2.35％).Approximately 9.99％(95％CI:8.47％-11.55％) of subjects had at least one indicator of kidney damage. Multivariate logistic regression revealed that albuminuria, hematuria, age and hyperuricemia were independently associated with reduced renal function. Hematuria and reduced renal function were independently associated with albuminuria. Albuminuria, reduced renal function and female were independently associated with hematuria. Conclusion In urban adult population over 35 years old of Urumqi, a big city in western China, the prevalence of CKD is 9.99％, the recognition is 2.44％ and the risk factors of CKD are similar to those of other domestic big cities and western developed countries.

Estrogen Receptor (ERalpha) is a member of superfamily of ligand-activated transcription factors which play critical roles in many biological processes. To screen novel modulators of ERalpha for drug development and biological function research, we developed a mammalian one-hybrid-based high-throughput screening model for ERalpha modulator. We cloned the ERalpha LBD gene from the total mRNA of fat tissue by RT-PCR and fused it with the GAL4 DNA binding domain of pBIND-GAL4 plasmid to construct a chimara expression plasmid pBIND-GAL4-Eralpha(LBD). The L02 cells was cotransfected with pBIND-GAL4-ERalpha(LBD) and a GAL4-responsive luciferase reporter plasmid pGL3-GAL4, and following treatment with test compounds for 24 h, the activities of luciferase were detected to evaluate the transactivities of ERalpha modulators. After manner optimizations of transfection conditions, Estradiol, an agonist control, induced the expression of luciferase in a dose-dependent with EC50 of 0.17 micromol/L, the maximum folds of induction was about 28.1. Tamoxifen, an antagonist control, efficiently suppressed the estradiol-mediated luciferase induction with EC50 of 0.10 micromol/L. Using this screening model, we discovered four ERalpha agonists from 2000 natural and synthetic compounds.
3T3-L1 Cells ; Animals ; Chimera ; metabolism ; DNA-Binding Proteins ; biosynthesis ; genetics ; Estrogen Receptor Modulators ; chemistry ; isolation & purification ; Estrogen Receptor alpha ; agonists ; Genes, Reporter ; genetics ; Genistein ; chemistry ; isolation & purification ; HeLa Cells ; Humans ; Luciferases ; genetics ; metabolism ; Mice ; Models, Chemical ; Saccharomyces cerevisiae Proteins ; biosynthesis ; genetics ; Transcription Factors ; biosynthesis ; genetics ; Transfection