BACKGROUND:Inducing factor and chondrogenic microenvironment is a primary factor, which influences chondrogenic differentiation and chondrogenesis of bone marrow-derived mesenchymal stem cells (MSCs). OBJECTIVE:To explore the feasibility of in vivo chondrogenesis by co-culture of bone marrow-derived MSCs and chondrocytes. DESIGN, TIME AND SETTING:A randomized controlled animal experiment was performed at Department of Pathology, Stomatological Hospital, Fourth Military Medical University of Chinese PLA between September 2004 and March 2005. MATERIALS:Fifteen New Zealand rabbits of clean grade were used for cell-scaffold construct transplantation. The rabbits were randomly divided into co-culture, chondrocyte, and bone marrow-derived MSC groups, with 5 rabbits in each group. Five neonatal New Zealand rabbits, aged 1-3 days, were used for isolation and culture of bone marrow-derived MSCs and chondrocytes. Polyglycolic acid (PGA) scaffold material (Shanghai Yikuo Company, China) has a fiber diameter of 15 μm, with an average interval of 150-200 μm, an interval porosity of 97% and 2-mm thickness. METHODS:In the co-culture group, bone marrow-derived MSCs and chondrocytes were mixed at a ratio of 3:1. The mixed cells were seeded onto a pre-wetted PGA scaffold (5 mm×5 mm )at the ultimate concentration of 6.0×1010 L-1. Dulbecco's modified Eagle's medium (DMEM) supplemented with fetal bovine serum was dropwise added to peripheral compound for 1 week of culture. In the chondrocyte, and bone marrow-derived MSC groups, chondrocytes and bone marrow-derived MSCs of the same ultimate concentration were seeded respectively onto the PGA scaffold. Then, the cell-scaffold constructs were transplanted into subcutaneous tissue of adult rabbits. MAIN OUTCOME MEASURES:Gross observation and hematoxylin-eosin & Masson staining of neo-cartilage were performed after in vivo culture for 8 weeks. RESULTS:Cell in all groups had a fine adhesion to the scaffold. In both co-culture and chondrocyte groups, the cell-scaffold constructs could maintain the original size and shape during in vivo culture and formed homogenous mature cartilage after 8 weeks of in vivo culture. Furthermore, the neo-cartilages in both groups were similar to each other in gross appearance and histological features. In the bone marrow-derived MSCs group, connective tissue rather than cartilage was found during in vivo culture. CONCLUSION:Chondrocytes can provide a chondrogenic microenvironment to induce a chondrogenic differentiation of bone marrow-derived MSCs and thus promote the chondrogenesis of bone marrow-derived MSCs in vivo.

Objective:To explore the clinical value on application of laparoscopic ultrasonography (Lap US) in the adnexal operation. Methods:Eleven patients including 7 cases of tubal pregnancy, 3 cases of teratoma of ovary and 1 case of endometrial cyst of ovary were examined by LapUS, then operated with laparoscopy.Results:The results showed that the modality presented is a big progress over the traditional operative management for adnexal diseas. Conclusions:It is a good approach for micro-surgery in the adnexal operation by laparoscopy.

Objective To determine the genetic background and evolutional route of macrolideresistance Streptococcus pneumoniae(MRSP)strains in Shanghai.Methods Forty-seven MRSP clinical isolates were genotyped by pulse field gel electrophoresis(PFGE)to detect the donal relationship of therm.Serotyping and muhilocus sequence typing(MIST)on 6 multi-resistant strains wag used to investigate the evolutional relationship between serum types and international epidemic strain among MRSP strains in Shanghai.Results There were 2 epidemical clones(type A 45%,type B 17%)in MRSP clinical isolates containing both the ermB and the mefE genes in Shanghai.MIST analysis showed that all 6 multi-resistant strains whose PFGE pattern was A type belonging to Asia clonal complex-CC236(Taiwan19F-14 clone).There was a novel ST-ST2116,which might derived from CC236 due to the recombination with alldic change.Conclusion,The high prevalence of erythromycin-resistant Streptococcus pneumoniae containing both the ermB and the mere genes in Shanghai is partly due to the clonal spread of a few mtdtidrng-resistant clones.

Objective To establish a new method, applying high resolution melt, to discriminate the VEB-3 hypotype from the clinical gram negative isolates. Methods From January to December 2003,292 consecutive and non-repetitive gram-negative bacteria producing VEB extended spectrum β-lactamase (ESBL) were collected. Extract the DNA of clinical gram negative isolates with phenol-chloroform. PCR was performed to amplify the VEB gene with the DNA being template. After that, we amplify the fragment of VEB gene containing the position 168. Then we detect the high resolution melt curve and analyze them. At last, we analyze the results of sequence and high resolution melt( HRM ). Results VEB-1 and VEB-3 gene are markedly different through HRM analysis. Conclusion It is accurately and quickly for us to identify the VEB-3 from other hypotype through the technology of HRM.

Objective To explore the influence of plasma matrix metalloproteinase-7 ( MMP-7 ) levels and genetic polymorphism of MMP-7 - 181 A/G on the stability of carotid plaque.Method According to carotid ultrasound examination, 503 patients with carotid atherosclerotic lesions were consecutively recruited and divided into vulnerable plaque group (n = 118) and stable plaque group (n = 385).Plasma MMP-7 levels were measured by enzyme-linked immunosorbent assay (ELISA), and MMP-7 -181 A/G genotypes were determined by polymerase chain reaction-restiction fragment length polymorphism (PCR-RFLP).Results Plasma MMP-7 levels in carotid vulnerable plaque group were significantly enhanced as compared to stable plaque group (t =5.49, P =0.00).The frequency of MMP-7 -181G allele in vulnerable plaque group was significantly higher than that in stable plaque group (11.4% vs 7.0% ,χ2 = 4.78, P= 0.029).Compared to AA genotype, the genotypes with - 181G allele (AG + GG) significantly increased susceptibility to carotid vulnerable plaque ( χ2 = 5.01, OR = 1.81, P = 0.025 ) .When further analyzing the relationship between genotype and plasma MMP-7 levels, no significant differences of plasma MMP-7 levels were observed between AA genotype and AG + GG genotype in stable plaque group.However, in vulnerable plaque group, plasma MMP-7 levels of AG + GG genotype were significantly higher than that of AA genotype( t = 2.62, P = 0.01).Conclusion The present findings suggest that plasma MMP-7 level may be a biomarker for carotid vulnerable plaque.Genetic polymorphism of - 181 A/G in MMP-7 promoter may affect the expression of MMP-7, and seems to be implicated in susceptibility to carotid vulnerable plaque.

Antimicrobial resistance is a serious problem for anti-infective treatment. Molecular technology can quickly, sensitively and accurately detect the mechanism of drug resistance of bacteria, improving the efficacy of anti-infection treatment and the level of infection control. The construction of quality assurance system is a guarantee for the effective application of molecular diagnostic technology in the detection of bacterial resistance. However, due to the complex mechanism of drug resistance, coupled with genetic mutations and other factors, there are problems such as false negatives, false positives, and inconsistency between mechanisms and phenotypes, there are certain restrictions on the application of molecular detection technology. With the development of molecular technology and deepening of people′s understanding of the drug resistance mechanism, the application of molecular diagnostic technology in the detection of bacterial resistance will be more widespread.

Objective To discuss the importance of data coding standards for the application of information technology , and put forward a method to solve the identification of complex medical materials in database structure design .Methods utilization of standardized coding , establishment of coding rules and a series of basic data dictionary , improvement of property fields and association dictionaries were used to regulate the generation , collection and processing of information , and to establish data standards for military medicine warehouse management information system .Results The design and implementation of data standards could be applicable for a variety of databases .Conclusion The establishment of standards and regulations of the data were critical to system design , develop-ment, implementation and even the successful operation , especially in management information systems , which had complex manage-ment form including property management , administration, and packaging assembly management .

Objective:To investigate the urine output threshold of acute kidney injury in patients with acute pancreatitis(AP) and to guide early fluid therapy.Methods:The clinical data of AP patients from Medical Information Mart for Intensive Care Ⅳ (MIMIC-Ⅳ) were collected. The 24-h urine output rate [24-h urine output·kg-1·24-h-1, 24-UR mL/ (kg·h) ] and 48-h urine output rate [48-h urine output·kg-1·48-h-1, 48-UR mL/ (kg·h) ] were calculated, and according to the occurrence of acute kidney injury within 7 days (7-AKI), AP patients were divided into the 7-AKI group and non-7-AKI group. The receiver operating characteristic (ROC) curve was drawn to evaluate the predictive value of 24-UR and 48-UR on 7-AKI in AP patients. 24-UR and 48-UR were grouped according to the optimal cut-off value obtained from the ROC curve. Logistic regression was used to analyze the risk factors of 7-AKI, and Kaplan-Meier (KM) survival curve was drawn to analyze the effect of 24-UR and 48-UR on in-hospital mortality of AP patients.Results:A total of 713 AP patients were included, ROC curve analysis showed that the area under the ROC curve (AUC) of 24-UR in predicting 7-AKI in AP patients was 0.76. Based on the maximum Youden index, the cut-off value of 24-UR was 0.795 mL/ (kg·h) , and the AUC of 48-UR was 0.78 and the cut-off value of 48-UR was 0.975 mL/ (kg·h) . Logistic regression analysis showed that 24-UR≤0.795 mL/ (kg·h) was an independent risk factor for 7-AKI compared with 24-UR>0.795 mL/ (kg·h) ( OR: 4.22, 95% CI:1.50-11.85, P=0.006). Similarly, compared with 48-UR>0.975 mL/ (kg·h) , 48-UR0.975 mL/ (kg·h) was an independent risk factor for 7-AKI ( OR: 3.75, 95% CI: 1.45-9.72, P=0.007). The KM survival curve showed that the cumulative in-hospital survival rate in the high 24-UR group was higher than that in the low 24-UR group. Conclusions:24-UR can be used to guide early fluid therapy in AP patients.

Objective:To evaluate the effect of invasive arterial blood pressure (IBP) monitoring on the prognosis of patients with sepsis.Methods:Patients with sepsis from the MIMIC-Ⅳ database were collected and divided into IBP and non-invasive blood pressure monitoring (NIBP) groups according to whether IBP monitoring was performed. Baseline variables that were considered clinically relevant or showed a univariate relationship with the outcome were entered into a multivariate logistic regression model as covariates.Propensity score matching(PSM) and inverse probability of treatment weighing(IPTW) were used to adjust confounders to ensure the robustness of findings.Subgroup analysis were conducted to evaluate the effect of differences in IBP onset and duration on outcome.Results:The 28-day mortality is lower in IBP group compared with NIBP group( OR=0.54, 95% CI 0.46-0.62, P<0.001), the conclusion maintain robust after PSM and IPTW.Then we conducted a series of logistic regression regarding to different initial IBP time(<24 h,24 h-48 h,>48 h) and the initial IBP time within 24 h showed the same results compared to primary outcoms( OR=0.42, 95% CI: 0.36-0.49, P<0.001). IBP duration varied (≤1day, ≤2days, ≤3days, ≤4days, >4days) all showed a statistically significant association with decreased 28-day mortality in the IBP group. Conclusions:IBP is associated with decreased 28-day mortality in patients with sepsis, and the optimal time of IBP is within 24 hours.

Objective To evaluate the changing pattern of antibiotic resistance inKlebsiella strains isolated from the patients in 19 hospitals across China based on the data from CHINET Antimicrobial Resistance Surveillance Program during the period from 2005 through 2014.Methods Kirby-Bauer disk diffusion and automated susceptibility testing methods were used to test the susceptibility ofKlebsiella isolates to the commonly used antibiotics. The results were interpreted according to the criteria of the Clinical and Laboratory Standards Institute (CLSI) Performance Standards for Antimicrobial Susceptibility Testing (CLSI-2014).Results A total of 61 406Klebsiella strains were identified between 2005 and 2014, includingK. pneumoniae (56 281 strains), K. oxytoca(4 779),Klebsiella pneumoniae subsp.Ozaenae (300) and otherKlebsiella species (46). Most (89.0%, 54 664/61 406) of theKlebsiella strains were isolated from inpatients, and 60.0% (36 835/61 406) were from respiratory tract speciems. About 16.7% (10 248/61 406) of the strains were isolated from pediatric patients aged 0-17 years and 83.3% (51 158/61 406) from adult patients. The prevalence ofKlebsiella spp. increased with time from 10.1% in 2005 to 14.3% in 2014. Based on the surveillance data during the 10-year period, we found a marked increase of resistance to imipenem (2.9% to 10.5%) and meropenem (2.8% to 13.4%) inKlebsiella spp. The prevalence of ESBLs-producing isolates inK. pneumoniae andK. oxytoca decreased from 39.0% in 2005 to 30.1% in 2014. The resistance to amikacin, ceftazidime, ciprolfoxacin, pipracillin-tazobactam and cefoperazone-sulbactam was on decline. The resistance rate to cefotaxime remained high about 49.5%. Carbapenem resistantance was identiifed in 5 796 (9.4%) of the isolates, including 5 492 strains ofK. pneumoniae and 280 strains ofK. oxytoca. Overall, 4 740 (7.8%) strains were identiifed as extensively-drug resistant (XDR), including 4 520 strains ofK. pneumoniae and 202 strains ofK. oxytoca. The carbapenem-resistant strains showed high (>60%) resistance rate to majority of the antimicrobial agents tested, but relatively low resistance to tigecycline (16.8%), amikacin (54.4%), and trimethoprim-sulfamethoxazole (55.5%).Conclusions During the 10-year period from 2005 through 2014, carbapenem resistance amongKlebsiella isolates has increased dramatically in the hospitals across China. The level of resistance to other antibiotics remains stable.