Objective:To investigate the effect of miRNA-1-3p (miR-1-3p) on expression of myocyte enhancer factor 2A (MEF2A) and the biological function of osteosarcoma cells.Methods:The tumor tissues and adjacent normal tissues of 20 patients with osteosarcoma who were clinically diagnosed in Shanxi Provincial Cancer Hospital from January 2019 to January 2020 were collected, and the expression of miR-1-3p in the samples was detected by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR). The expression of miR-1-3p in osteosarcoma cell lines U2-OS, SAOS-2, MG63, SW1353 and human normal osteoblast cell line hFOB1.19 was detected by qRT-PCR, then the cell line with the lowest expression of miR-1-3p was selected for follow-up experiments. An overexpression miR-1-3p vector was constructed (miR-1-3p mimcs). The miR-1-3p overexpression group was transfected with miR-1-3p mimcs, and the control group was transfected with empty vector (miR-1-3p nc). CCK-8 method was used to detect the proliferation activity of cells; flow cytometry was used to detect the changes of cell apoptosis and cell cycle. miRwalk database was used to predict the miR-1-3p target gene, and the target gene was verified by dual-luciferase reporter gene assay; Western blot was used to detect the expression of MEF2A protein in cells of each group.Results:Compared with adjacent tissues, the expression of miR-1-3p in osteosarcoma tissues was down-regulated (0.31±0.14 vs. 0.62±0.21), and the difference was statistically significant ( t = 5.31, P<0.01). The expression of miR-1-3p in U2-OS cells was the lowest; compared with the control group, the proliferation activity of U2-OS cells was inhibited in miR-1-3p overexpression group (48 h absorbance value 0.56±0.01 vs. 0.77±0.03, t = 2.77, P<0.01; 72 h absorbance value 0.87±0.02 vs. 1.40±0.03, t = 2.93, P<0.01); G 1/S cell cycle arrest increased [G 1 phase (38.24±0.55)% vs. (32.11±0.80)%, t = 9.27, P = 0.01; S phase (61.24±0.90)% vs. (67.78±0.83)%, t = 7.52, P = 0.02]; early apoptotic rate increased [(11.20±0.12)% vs. (1.50±0.12)%, t = 2.91, P<0.05], miRwalk database predicted that the miR-1-3p target gene was MEF2A. The result of dual-luciferase reporter gene assay showed that miR-1-3p bound to MEF2A 3'UTR, and the luciferase activity of U2-OS cells in miR-1-3p overexpression group was lower than that in the control group (renilla luciferase/firefly luciferase activity ratio 0.53±0.06 vs. 1.00±0.04, t = 4.04, P < 0.05). Western blot showed that the expression of MEF2A protein in U2-OS cells of miR-1-3p overexpression group was lower than that of the control group (protein relative expression 0.41±0.14 vs. 0.77±0.12, t = 3.93, P < 0.05). Conclusions:The low expression of miR-1-3p may be associated with the proliferation, apoptosis and cycle changes of osteosarcoma cells. miR-1-3p can negatively regulate the expression of MEF2A protein and regulate the occurrence and development of osteosarcoma.

Objective To probe the proper management of dural tear sustained during operation on the lumbar spine and cerebrospinal fluid leakage. Methods Sixty-eight patients sustained dural tears, which were repaired during the operation. According to the type of the dural tear, different ways was used to treat the cerebrospinal fluid leakage.Results Fifteen patients of the sixty-eight had cerebrospinal fluid leakage. None have any longterm deleterious effects. Conclusions Repair the dura during the operation was the best way to treat the dural tears, and closed subarachnoid drainage can successfully treat the cerebrospinal fluid leakage.

Objective To prepare and optimize fibrin-gel-coated vaneomycin alginate beads (FG-Vanco-AB)and investigate their possible use in treatment of osteomyelitis or prevention of infection.Methods Vancomycin alginate beads were produced by dropping vancomycin and alginate mixed liquor into calcium chloride solution.Beads including high vancomycin content were prepared and chosen by optimizing different concentrations of vancomycin solution and alginate solution.These beads were coated with fibrin gel formed by different concentrations of fibrin and the same concentration thrombin.The optimized beads were selected based on available release time,when vancomycin in medium could kill Staphylococcus aureus(ATCC25923). Results Higher content of vancomycin in bead resulted in increase of vancomycin concentration and alginate concentration in mixed liquid.The highest vancomycin content beads were prepared by 16%alginate and 50 mr/ml vancomycin,up to(27.36±0.90)%.The further results showed that vancomycin concentrations from beads coated with fibrin at 75 mg/ml and thrombin at 400 IU/ml could kill Staphylococcus aureus and remained above the breakpoint sensitivity for 19 days.Conclusion The available release time is prolonged,and the possibility of clinical use is conspicuously increased after vancomycin beads are optimized by adjusting the rate of mixed component and fibrin gel coat.

Objective To compare the effect of pancreatic duct stent internal versus external drainage in the prevention of postoperative complications after pancreaticoduodenectomy through the method of Meta analysis.Methods PubMed,Embase and the Cochrane Library,were searched for randomized controlled trials (RCTs) concerning pancreatic duct stent in the prevention of postoperative complications after pancreaticoduodenectomy.All these databases were searched from their establishment to March 31,2015.The data was reviewed and extracted by two investigators independently.Then,the Cochrane network RevMan 5.3 software was used for statistic analysis.Results As a result,this meta analysis has got 3 RCTs,including 362 participants.The outcomes in our study design were classified as major and minor one.The former was the outcomes of the major postoperative complications,like postoperative pancreatic fistula and delayed gastric emptying.The minor outcome were postoperative morbidity,mortality and intestinal obstruction.The results of meta analysis were:(1) Postoperative total pancreatic fistula rate (A/B/C):three studies showed a statistic difference between the internal and external drainage groups (OR =0.59,95％CI:0.36-0.97,P =0.04).(2) Postoperative pancreatic fistula rate (B/C):three studies showed a statistic difference between the internal and external drainage groups (OR =0.44,95％ CI:0.20-0.97,P =0.04).(3) Postoperative incidence rate of delayed gastric emptying:three studies showed a certain statistic difference between the internal and external drainage groups (OR =0.42,95 ％ CI:0.23-0.79,P =0.007).(4) Post-operative incidence rate of total mortality:three studies showed no certain statistic difference between the internal and external drainage groups (OR =0.81,95 ％ CI:0.23-2.86,P =0.74).(5) As for the postoperative incidence rate of total complications and intestinal obstruction,the heterogeneity was bigger than 50％.So we made an analysis of the cause of heterogeneity.We deduced that it may be caused by the different and complicated perioperative management.Then,we used the random effect model rather than the fixed effect model to make a quantitative analysis.No statistical difference was found eventually in both this two marks.Conclusions By comparing the outcomes in both internal and external drainage groups,we found pancreatic duct stent external drainage could effectively decrease the incidence rate of postoperative pancreatic fistula rate and delayed gastric emptying.But when the limit studies and sample size considered,this conclusion still need to be certificated with more high-quality clinical research.

ObjectiveTo discuss the selection of surgical approach,operative methods,and stability of reconstitution of affection of cervicothoracic junction.MethodsFrom January 2001 to February 2009,86cases with affection of cervicothoracic junction were treated surgically.The mean age of patients at the time of surgery was 43.1 years (range,17-70).Fifty-seven patients were treated with anterior approach (fixation with autologous bone grafts was done in 38 patients,Cage fixation in 5,titanium mesh in 14),21 with posterior approach(the fixation of lateral mass screw combined with pedicle screw was used in 12 patients,fixation with pedicle screw in 9),and 8 with anterior combined posterior approach.The neurological function of 53 cases of injury of cervicothoracic junction was assessed by American Spinal Injury Association (ASIA) criteria,and the rest was assessed by Japanese Orthopaedic Association(JOA) criteria; bone arthrodesis and restoring lordosis of cervical spine were assessed by Bohlman radiographic criteria.ResultsSixty-nine cases were followed up for an average of 12.4 months(range,3-45).ASIA score increased from 1.8 preoperatively to 2.3 postoperatively,and JOA score increased from 10.3 preoperatively to 12.8 postoperatively.Bone fusion reached in all patients,and lordosis of the cervical spine of 62 cases was restored.One case with respiratory dysfunction,1 case with cerebrospinal fluid leakage,2 cases with hoarseness,and 1 case with loose lateral mass screw were found after the surgery.ConclusionThe advantages of surgical reconstitution of the cervicothoracic junction included promoting recovery of neurological function,restoring the alignment and lordosis of the cervical spine,decreasing the rate of complication,which were dependent on the suitable surgical indications,surgical approach,and way of surgical reconstitution.The selection of reconstituted methods depends on disease,lesion site,type and degree of injury,experience of doctor,decompression and spinal stabilization synthetically.

Objective To study the effects of CYP3A5 * 1 and CYP3A5 * 3 genotypes on tacrolimus dose requirement.Method We tested archival peripheral blood of 69 kidney recipients for CYP3A5 genotyping by polymerase chain reaction.The dose,blood concentrations and dose-normalized blood concentrations of tacrolimus were measured at 1st and 2nd month after the renal transplantation.Result There were 6 cases of CYP3A5 * 1/* 1 (8.7％),22 cases of CYP3A5 * 1/* 3 (31.9％),and 41 cases of CYP3A5 * 3/* 3 (59.4％).At 1st and 2nd month after the renal transplantation,the carriers of CYP3A5 * 1 genotype had a higher mean tacrolimus dose than CYP3A5 * 3/* 3 genotye (both P＜0.000 1),and those of CYP3A5 * 1 genotype had lower mean tacrolimus concentrations than CYP3A5 * 3/* 3 genotye (P=0.020 8,and P =0.019 1 respectively).Meanwhile,the carriers of CYP3A5 * 1 genotype had lower dose-normalized blood concentrations of tacrolimus than CYP3A5 * 3/* 3 genotye (P＜0.000 1) at 1st month after the renal transplantation,as well as at 2nd month after the renal transplantation (P =0.0191).Hepatic and renal function showed no significant effect on tacrolimus dose adjusted concentration at 1st and 2nd month after transplantation.Gender did not show a significant impact on tacrolimus dose.Conclusion CYP3A5 * 1 carriers needhigher tacrolimus dose than CYP3A5 * 3 homozygote to achieve the target blood concentration.CYP3A5 genotyping is a new approach for detecting tacrolimus dose requirement in kidney recipients.

Objective To determine the conversion equation for X(copies/ml, lg) quantity values of domestic HCV RNA quantitative fluorescence amplification assays approved by SFDA and Y( IU/ml, lg)reference values of standard substance. Methods The second generation WHO International Standard (NIBSC code:96/798) was mixed with human AB blood type serum to create 7 different dilutions which included 100 000, 50 000, 25 000, 10 000, 5 000, 2 500 and 1 000 IU/ml. Two different batches of each three domestic hepatitis C virus RNA real-time fluorescence quantitative PCR assays and 2 different batches of each assay were employed to detect the 7 different concentration samples with real-time PCR. Each test was performed 4 times repeatedly. Results The correlations between X( copies/ml,lg) values of domestic HCV RNA assays and Y(IU/ml,lg) reference values of standard substance were as follow,Assay A:Y =0. 902 0 X+0.284 9,R2 =0.953 3,P＜0. 01,n =56;Assay B: Y=0. 875 7 X +0.562 4,R2 =0.956 5,P＜0.01,n =56; Assay C: Y = 0. 843 8 X + 0. 560 5, R2 = 0. 945 8, P ＜ 0. 01, n = 56. Conclusions All the conversion equations are different among the quantity value of three assays and the reference values of standard substance, that suggests it is necessary to perform more stringent traceability analysis for the quantity values of 3 assays. Through standardizing the quantity values preliminarily, the conversion equation can enhance the comparability between the quantity values of different assays, and provide a standard of HCV RNA virus load detection for clinical diagnosis and treatment monitoring of HCV infection.

Objective To explore the clinical effect of the open and laparoscopic surgery in the treatment of ectopic pregnancy to retain the clinical efficacy of reproductive function analysis. Methods Four hundred and fifty-six patients with ectopic pregnancy were selected as our subjects. Two hundred and eleven cases were served as laparotomy group( open),and 245 cases were laparoscopic group( laparoscopic). The data of intraoperative blood loss,operative time,postoperative anal exhaust time,average hospital stay,tubal patency rate,intrauterine pregnancy rate and again repeat ectopic pregnancy rate were recorded. Results Patients in two groups were successfully completed surgery. The operative time,intraoperative blood loss,postoperative anal exhaust time,average hospitalization time were(55. 1 ± 13. 5)min,(63. 5 ± 18. 3)ml,(25. 7 ± 5. 6)h,(6. 1 ± 2. 0)d respectively in laparotomy group,and(41. 3 ± 15. 5)min,(41. 1 ± 13. 3)ml,(13. 5 ± 5. 1)h,(3. 6 ± 1. 4)d respectively in laparoscopic group,and the differences were significant( t =2. 045,2. 263,3. 131, 3. 152,P﹤0. 05). The tubal patency rate,intrauterine pregnancy,ectopic pregnancy rate in laparotomy group were 81. 0%( 171/211 ),59. 7%( 126/211 ) and 26. 5%( 56/211 ) respectively,and 75. 5%( 185/245 ), 53. 5%( 131/245 ) and 22. 9%( 56/245 ) respectively in laparoscopic group,and the differences were not significant(χ2 =2. 254,2. 130,1. 242;P ﹥0. 05 ). Conclusion Laparoscopic surgery in the treatment of ectopic pregnancy is superior to laparotomy in terms of small trauma,less bleeding,faster recovery,shorter hospitalization time,and postoperative tubal patency rate.

Objective To analyze the metabolic profile in serum between normal and orthotopic xenograft nude mice burdened with three human pancreatic cancer cell lines,which were differentiated differently.Methods Human pancreatic cancer lines SW1990,BxPC-3 and Panc-1 were subcutaneously injected into the nude mice,respectively.When the tumor volume reached 1.0 cm3,the nude mice were euthanized and the tumor tissues were removed and implanted to the pancreas to establish the orthotopic xenograft mice model.The serum from three orthotopic xenograft tumor nude mice and the normal controls were collected and then analyzed by 1H nuclear magnetic resonance spectroscopy.Results The three orthotopic xenograft nude mice models were successfully established.In SW1990,BxPC-3 and Panc-1 group,the orthotopic xenograft tumor formation rate was 79％ (11/14),93％ (13/14) and 86％ (12/14),while the mortality was 7％ (1/14),0 and 7％ (1/14),respectively.Compared with control group,the content of metabolites in the serum of orthotopic xenograft tumor nude mice was increased including creatine,alanine,glutamine,1-methylhistidine,isoleucine,lactate,phenylalanine,tryptophan and valine,but the glycerolphosphocholine (GPC) and glucose levels were reduced.As the tumors progressed to be more malignant,the content of valine and isoleucine tended to increase.Conclusions The establishment of the orthotopic implantation tumor nude mice model was stable and reliable with high tumor formation rate.Obvious metabolic differences of glucose,lipid and amino acids were observed between normal and human pancreatic cancer tumor burdening nude mice models.The common metabolic features identified in all three nude mice models burdened with human pancreatic cancer could be used as the potential markers for diagnosing human pancreatic cancer.

Objective To investigate the clinical value of serum metabonomic profile of pancreatic cancer using nuclear magnetic resonance (NMR)-based metabonomics.Methods The retrospective case-control study was adopted.The clinical data of 23 patients with pancreatic cancer (PC group) and 16 healthy volunteers (control group) who were admitted to the Fujian Medical University Union Hospital between December 2013 and December 2014 were collected.The serum of the 2 groups was measured by 1H NMR spectroscopy.Multivariate statistical analyses were performed to identify the characteristic metabolites in the 2 groups,including principal component analysis (PCA),partial least squares discriminant analysis (PLS-DA) and orthogonal partial least squares discriminant analysis (OPLS-DA).Observation indicators included:(1) multivariate statistical analysis of serum metabonomic profile,results of PCA,PLS-DA and OPLS-DA,(2) screening of metabolites.Measurement data with normal distribution were presented as x ± s.The comparison between groups was evaluated with the t test.The count data were analyzed using the chi-square test.Results (1) The multivariate statistical analysis of serum metabonomic profile:results of PCA showed that expression rates of principal component 1 (PC1) and principal component 2 (PC2) to original data were 54.9％ and 23.5％,with both cumulative contribution rate of 78.4％.Results of PLS-DA showed that the separative trend between PC group and control group was appeared,and variance of X and Y matrixes and predictive value were 0.254,0.816 and 0.385.Results of OPLS-DA showed that the differences of samples between the 2 groups were further increased,and differential metabolites were screened according to the distinction of scores between the 2 groups,value of R2X,R2Y and Q2 was 0.254,0.816 and 0.433.(2) Screening of metabolites:35 serum metabolites were detected in the 2 groups.Compared with the control group,levels of 3-hydroxybuyarate,citrate,formate,glutamate,isoleucine,methionine and phenylalanine in the PC group were elevated (r =0.524,0.511,0.656,0.566,0.503,0.498,0.648,P ＜0.05),and levels of 3-methylhistidine,alanine,glutamine,LDL and VLDL in the PC group were decreased (r =-0.607,-0.508,-0.560,-0.568,-0.559,P ＜ 0.05).Conclusions Compared with healthy controls,several amino acids,citrate and lipoproteins demonstrate the metabolic differences in the serum of patients with pancreatic cancer.NMR based metabonomic profile technology can distinguish the difference of serum metabolites between patients with pancreatic cancer and healthy controls.NMR based metabonomic technology may be a promising method for the diagnosis of pancreatic cancer.