Objective To investigate the changes of plasma microRNA-223(miR-223) and HMGB-1 in pediatric sepsis patients.Methods There were 49 children with sepsis enrolled in the study (sepsis group),severe sepsis group (n=25) and general group (n=24). Meanwhile, 50 healthy children (normal control group) were selected as control group. The expression levels of plasma miR-223and HMGB-1 (high mobility group box 1) were detected. The predictive values of miR-223and HMGB-1 in plasma of children with sepsis were evaluated by receiver operatingcharacteristic (ROC) curve.Results The plasma miR-223 and HMGB-1 expression levels in severe sepsis group and general group were up-regulated compared with those in the normal control group (F=63.02, 76.32,P<0.05). The area under ROC curve of miR-223,HMGB-1 predicting sepsis were 0.904 (95%CI 0.821-0.998), 0.748 (95%CI: 0.625-0.903). There was positive correlation between miR-223 and HMGB-1 (r=3.532, P<0.05). Conclusions The expression levels of plasma miR-223 in children with sepsis are signiifcantly up-regulated, which can be used as early diagnostic markers to relfect the severity of inlfammation in some degree.

Objective: To establish a method for determining the contents of hesperidin and arctiin in the root of Arctium lappa L.from different habitats.Methods: The chromatographic separation was performed on an Acquity UPLC BEH C18 column (50 mm×2.1 mm, 1.7 μm).The mobile phase consisted of acetonitrile (A)-0.1% formic acid (B) with gradient elution at the flow rate of 0.3 ml·min-1.The detection wavelength was 280 nm, and the temperature of column was 25℃.Results: The linear range of hesperidin and arctiin was 4.950-29.700 μg·ml-1 and 6.250-37.500 μg·ml-1(r=0.999 9), respectively.The average recovery of hesperidin and arctiin was 99.0% and 98.5%, respectively.Conclusion: The developed method is sensitive, rapid and accurate, which can be used for the quality control of the root,of Arctium lappa L.

Objective To evaluate the necessity and practicability of optimizing the management of type 2 diabetes mellitus in community after acquiring the peer education and the music therapy to their physical and mental issues and sleep problems. Methods Totally 179 type 2 diabetes mellitus patients who were followed up in Ruijin 2nd community health service center, the random numbers table used to randomize the patients into 2 groups:control group ( 97 cases) and experiment group ( 96 cases), the conventional treatment was used in control group. Besides the conventional treatment measures, the peer support was used by patients in the experiment group. In the second step, 45 patients were met the inclusion and the exclusion criteria, the random numbers table used to randomize the patients into 2 groups:the multiple intervention group (22 cases) and the conventional treatment group (23 cases), the multiple intervention included the music therapy, the peer support and the sleep health education, the conventional treatment included the conventional treatment and the sleep health education. The t test was used to compare the patient's HbA1c and other quantitative data in two groups of patients after the intervention. Results In the first stage of research, compared with the control group patients, the patients 'HbA1c in intervention group was significantly improvement after 6 months(7.26%±1.37%vs.7.53%±1.63%,t=2.148, P<0.05),besides, the intervention group individuals achieved significant improvement in diabetic self-management behaviors and self-efficacy after 6 months, and the improvement in self-efficacy of peer support group was significant different compared with routinely educated patients(104.09±16.40 vs.110.96± 13.86,t=2.120,P<0.05), and the PHQ-9(5.95 ± 4.02 vs.2.55 ± 1.67,t=2.630,P<0.05)between the two group had significant difference, while no improvement was found in PSQI, BMI, and WHR between intervention group and control group. Conclusions Peer support could improve the blood glucose metabolism in patients with type 2 diabetes. With the effect of yoga music and physical exercises, peer support can improve the quality of sleep and decrease depression in T2DM patients, who also have sleep disorders and mild depression.

BACKGROUND: Key point for subculture of human embryonic stem cells (hESCs) is to inhibit spontaneous differentiation and make sure totipotency of cells. Although mouse embryonic fibroblasts (MEF) or human foreskin fibroblasts (HFF) used as the feeder layer can maintain undifferentiated state of embryonic stem cells, cell clone is still imperfect and parallel arranged. OBJECTIVE: To establish mixed feeder layer of mouse embryonic fibroblasts plus human foreskin fibroblasts and to observe the hESCs growth.DESIGN: Multi-sample observation and comparison.SETTING: Medical Center of Reproduction, the Affiliated Hospital of Hainan Medical College. MATERIALS: This study was performed at the Medical Center of Reproduction, the Affiliated Hospital of Hainan Medical College from April 2006 to July 2007. Foreskin was derived from the children who underwent circumcision and came from Urinary Surgery, the Affiliated Hospital of Haihan Medical College. The children's family members provided the informed consent, and the experiment received confirmed consent from the local ethic committee. hESCs line HN-1 was separated from human blastula. Eleven 12.5-14.5-day-old fetal mice of clean grade were selected in this study. The experimental animals were disposed according to ethical criteria. METHODS: Heads, four extremities, and viscera were removed from fetal mice under anesthesia. Subsequently, cell suspension was prepared using routine trypsinase digestion and inoculated. When cells were cultured in confluent monolayer, some primary cells were frozen, processed with mitocin-C for 2.0-3.0 hours, and inoculated based on the density of 1×108 L-1 in gelatin-coated dish, I.e., MEF feeder layer. The HFF separation and culture and the preparation of HFF feeder layer were the same as above-mentioned processing. In addition, the two fibroblasts were respectively counted and mixed together according to the ratios of 1∶0, 3∶1, 1∶1, 1∶3, and 0:1. And then, the mixture was inoculated based on the density of 1×108 L-1 in gelatin-coated dish, I.e., mixed feeder layer. The growth of subcultured hESCs in vitro was observed in three different feeder layers, and hESCs in the mixed feeder layer underwent alkaline phosphatase test, OCT-4 expression immunohistochemical measurement, and OCT-4 and telomerase mRNA expression RT-PCR detection. Finally, differentiation in vitro of hESCs was observed after removing the feeder layer.MAIN OUTCOME MEASURES: ① Growth of hESCs in the three different feeder layers; ② Growth of hESCs in the mixed feeder layer based on different mixed ratios; ③ undifferentiated state of hESCs in the mixed feeder layer; ④ differentiation in vitro.RESULTS: ① hESCs clone in the MEF and HFF feeder layers was thin and flat and imperfect, but hESCs clone in the mixed feeder layer was perfect and thick and solid. Apparently, the clone form in the mixed feeder layer was superior to MEF and HFF feeder layers. ② When MEF and HFF was mixed together according to the ratio of 1∶1, hESCs grew in apparent accumulation; clone border was clear; eminentia was apparent and perfect. However, there were no changes according to the ratio of 1∶3. The ratio of 1∶1 was superior to the ratios of 1∶0, 3∶1, and 0∶1. ③ Alkaline phosphatase staining and OCT-4 antigen expression were strongly positive. Specific straps of OCT-4 and telomerase mRNA expression were observed at 200-300 bp and 300-400 bp, respectively. ④ Embryoid bodies were formed. hESCs could differentiate into multi-morphological cells after attachment.CONCLUSION: ① The mixed feeder layer may well support in vitro subculture of hESCs to acquire excellent clone form compared to MEF or HFF feeder layer. ② The mixture of MEF and HFF has excellent effect according to the ratio of 1∶1.

A) in COL1A1 gene resulting in OI in a Chinese family. The detailed molecular and clinical features will be useful for extending the evidence for genetic and phenotypic heterogeneity in OI and exploring the phenotype-genotype correlations in OI.

Objective To evaluate the capability of multilocus variable-number tandem-repeat ( VNTR) analysis ( MLVA) and pulsed-field gel electrophoresis ( PFGE) for genotyping Salmonella enterica serovar Typhi (S.Typhi) isolates.Methods Five polymorphic VNTRs (SAL02,SAL11,SAL16,SAL20, and TR4699 ) that were observed in S.Typhi strains from previous studies were selected to establish MLVA . Twenty-one epidemiologically unrelated S.Typhi strains that were isolated from Shenzhen ,China from 2002 to 2007 were genotyped by the established MLVA , and the results were compared with those by PFGE . Results The Simpson′s index of diversity ( D value ) for all five different VNTRs ranged from 0.838 to 0.943 .A total of 19 MLVA profiles and 19 PFGE profiles were found , respectively .The D value for both MLVA and PFGE were 0.99 and the test results from two analyses were identical .Conclusion The five polymorphic VNTRs analysis could be used as an alternative typing scheme for epidemiologic investigation of S.Typhi infection .

Objective To investigate the prevalence and molecular characteristics of the extended -spectrum β-lactamase ( ESBL) and AmpC enzyme-producing Proteus mirabilis ( P.mirabilis) strains isola-ted in Shenzhen People′s Hospital.Methods The production of ESBLs and AmpC enzymes by P.mirabilis isolates were detected by a screening and confirmatory test for ESBLs and AmpC disk test , respectively .The PCR assays followed by DNA sequencing of the products were employed to analyze the multiple genes inclu -ding the ESBLs genes, AmpC genes, insertion sequences (ISs) upstream of the ESBLs or AmpC genes, plasmid -mediated quinolone resistance ( PMQR ) determinants , quinolone resistance-determining region (QRDR) genes , the integrase genes, and class1 integron cassette.The epidemiological analysis of the iso-lates was performed by pulsed field gel electrophoresis .Results There were 130 P.mirabilis clinical iso-lates collected from Shenzhen People′s Hospital in China during the year 2004 to 2010.Among them, 13 isolates (10%) produced ESBLs, that accounted for 0%-9.1%in the year 2004-2009 and up to 29.4%in 2010, and 3 isolates (2.3%) produced AmpC enzymes.The predominant genotype of ESBLs -producing isolateswas b al CTX-M-14(n=7), followed by blaCTX-M-65(n=3), blaCTX-M-55(n=1), blaCTX-M-24(n=1) and blaPER-1 (n =1).The clinical isolate of PER-1-producing P.mirabilis was reported for the first time in China.Twoisolates carried an AmpC β-lactamase gene of blaCMY-2 and one isolate carried an unidentified AmpC gene .ISEcp1 located upstream of blaCTX-M and blaCMY-2 were detected in 91.7% (11/12) of CTX-M-producing isolatesand one CMY-2-producing isolate, respectively.ISPa12 was present upstream of blaPER-1 in one studiedisolate.Approximately 66.7% (10/15) of ESBL and /or AmpC-producing isolates harbored PMQR genes including2 carrying qnrD, 5 carrying aac-Ib-cr and 3 carrying both qnrD and aac-Ib-cr.Twelve ESBL and /orAmpC-producers with high level of resistance to ciprofloxacin carried the similar mutation profiles of S 83I inGyrA, S80I or S80R in ParC and among them, six strains showed E466D mutation in GyrB.Approximately86.7% (13/15) of ESBL and/or AmpC-producing isolates carried class 1 integron.Fourteen PFGE typeswere observed among 15 ESBL and/or AmpC-producers.Conclusion The prevalence of CTX-M β-lactamasesin P.mirabilis isolates contributed to the increased resistance to extended -spectrum cephalosporins.The qnrD and/or aac-Ib-cr genes were detected among the most of ESBL and /or AmpC-producing P.mirabilis clinical isolates.

Objective To study effects ofXiangsha Liujunzi Decoction (XSLJZ) on gastric emptying function, pepsin activity and expression of HIF-1α gene and protein of gastric mucosa in chronic atrophic gastritis (CAG) of rats with deficiency of spleen and stomach.Methods Rats were randomly divided into normal group and model group. The models of CAG rats with deficiency of spleen and stomach type were induced by synthetic methods. After the modeling, models rats were divided into model group, positive control group and XSLJZ high-, medium-, and low-dose groups. Normal group and model group were given 10 mL/kg distilled water every day for gavage; XSLJZ high-, medium-, and low-dose groups were given 24, 12, and 6 g/kg XSLJZ Decoction for gavage; positive control group was given 0.30 g/kg Vatacoenayme for gavage, for successive 120 d. Gastric emptying function and pepsin activity were detected, and HIF-1α gene and protein expression in gastric mucosa were detected by RT-qPCR and IHC. Results Compared with the normal group, the gastric emptying function and pepsin activity in the model group were much lower (P<0.01); expressions of HIF-1α gene and protein in gastric tissues was much higher (P<0.01). Compared with model group, XSLJZ could increase the gastric emptying function and pepsin activity significantly (P<0.05), and decrease expressions of HIF-1α gene and protein (P<0.05).Conclusion XSLJZ has functions of improving the gastric emptying function, promoting pepsin activity, and reducing expressions of HIF-1α gene and protein.

Objective To discuss the effects of electroacupuncture intervention in different acupoints of rats models with diarrhea type irritable bowel syndrome (D-IBS).Methods The rat models were established with acute and chronic stimulation. The rats were divided into the normal control group, model group, “Taichong” (LR3) + “Zusanli” (ST36) group, and “Tianshu” (ST25) + “Zusanli” (ST36) group. Electroacupuncture was applied to bilateral corresponding acupoints for 20 minutes, every other day for 7 times. The physical signs of each rat were observed. The contents of 5-HT, NE and CGRP in the rats’ plasma were detected by sandwich ELISA. Results Compared with the normal group, the contents of 5-HT and NE in the model group were obviously higher and the content of CGRP decreased (P<0.05). Compared with the model group, the contents of 5-HT and NE in all treatment groups were lower and the content of CGRP increased (P<0.05). There was no difference between the “Taichong” (LR3) + “Zusanli” (ST36) and “Tianshu” (ST25) + “Zusanli” (ST36) groups.Conclusion The mechanism of electroacupuncture may be realized through lowering the contents of 5-HT and NE in the model rats and increasing the content of CGRP to treat D-IBS.

ObjectiveTo perform pharmacology problem-based learning (PBL) and evaluate its effects.MethodsPBL was performed for the clinical medicine class of grade 2007 and the satisfactory degree of students to teaching effects was observed with questionnaire. Results The students thought that PBL teaching had substantial contents and proper schedule and increased learning interest. Students' participating degree, mutual communication and controlling discussion procedure were fine,which reached the expected learning objective. ConclusionsThe effects of PBL teaching were excellent and most of our students could accept it.