BACKGROUND:Classic media and fetal bovine serum are commonly used in the culture of umbilical cord mesenchymal stem cells, but the potential risk of serum culture limits its clinical application. OBJECTIVE:To use human platelet lysate alternative to fetal bovine serum for large-scale production of mesenchymal stem cells for therapeutic applications. METHODS:Human platelet lysate was prepared by repeated freezing and thawing, centrifugation, filtration, and concentration. Human umbilical cord mesenchymal stem cells were cultured in Iscove’s Modified Dulbecco’s Medium containing 5%concentrated platelet lysate (experimental group) or 10%fetal bovine serum (control group). After separation and enzymatic digestion, human umbilical cord mesenchymal stem cells were subcultured at a concentration of 3 000/cm2 up to the fifth generation. Then, cellmorphology and diameter, immune phenotype, osteogenic and adipogenic differentiation, and cloning efficiency were detected and compared between two groups. RESULTS AND CONCLUSION:Human umbilical cord mesenchymal stem cells cultivated in human platelet lysate-supplemented media showed a smal er size and more elongated morphology than those in fetal bovine serum-supplemented media. Colony forming unit-fibroblast analyses further showed no significant differences in colony efficiency. Human umbilical cord mesenchymal stem cells cultivated in human platelet lysate-supplemented media showed an increase of proliferation capacity;whereas, similar immunophenotypes remained in the two groups. In vitro assays revealed intact differentiation potential. Moreover, human umbilical cord mesenchymal stem cells cultivated in human platelet lysate-supplemented media showed a significantly higher capacity to differentiate towards osteocytes, indicating human platelet lysate is an alternative to fetal bovine serum for low-density production of mesenchymal stem cells for therapeutic applications.

Objective To investigate the serum level of placental growth factor (PLGF) and explore its relationship with left ventricular structure and function in chronic kidney disease (CKD) patients.Methods Seventy-two non-dialysis CKD patients and sixteen age-and sex-matched healthy controls were included in this study.Serum PLGF level was measured by ELISA.Cardiac structure and function were assessed by two dimensional echocardiography.Results (1)The serum level of PLGF in CKD patients[3.32(2.97,19.77) ng/L] was significantly higher compared to the healthy controls [2.33(2.27,2.49) ng/L] (P ＜ 0.01).It progressively increased with the decline of renal function (P＜ 0.05/6).(2)The interventricular septum (IVS),left ventricular posterior wall (LVPW) was significantly higher while the ejection fraction was significantly lower in CKD patients.(3)The serum PLGF level was higher in patients with left ventricular hypertrophy (LVH) than those without LVH [19.05(3.31,21.05) ng/L vs 2.99(2.60,3.32) ng/L,P ＜ 0.05].The prevalence of LVH in the group above median PLGF level was significantly higher than that in the group below the median PLGF level (70％ vs 18％,P ＜ 0.01).(4)PLGF level was positively correlated with left ventricular mass index (LVMI),systolic pressure,diastolic pressure,24 h urine protein,Scr,UA,BUN,iPTH,the history of high blood pressure and was negatively correlated with LVEF,hemoglobin,albumin,eGFR (P ＜ 0.05).Multiple regression results showed that UA,Scr,LVEF,Hb were associated with PLGF level independently (P ＜ 0.01).Conclusions CKD patients have elevated level of PLGF.It has a relationship with cardiac structure and function.It may participate in the occurrence of cardiovascular events.

Objective:To explore the role of the fat mass and obesity-associated protein (FTO) in human renal tubular epithelial cells (HK-2) suffering ischemia-reperfusion injury (IRI).Methods:The in vitro IRI mo-del was established in HK-2 cells by induction with antimycin A, A23187 and 2-deoxy-D-glucose.The cells were divided into control group and ischemia-reperfusion group (I/R group). The mRNA and protein expressions of FTO, B-cell lymphoma / leukemia 2(Bcl-2)-associated X(Bax), Bcl-2 and cleaved cysteinyl aspartate specific proteinase(cleaved Caspase-3) in HK-2 cells before and after IRI were detected by real-time fluorescent quantitative PCR(qPCR) and Western blot, respectively.Cell apoptosis was measured using flow cytometry.The level ofe N 6-methy-ladenosine (m 6A) RNA was detected by colorimetry. Results:(1) The mRNA expressions of FTO (0.15±0.05 vs.1.00±0.23) and Bcl-2 (0.14±0.07 vs.1.02±0.25) in I/R group were significantly lower than those in control group; While those of Bax (3.10±0.35 vs.1.00±0.13) and cleaved Caspase-3 (4.21±0.56 vs.1.00±0.09) were significantly higher ( t=6.28, 5.84, -9.83, and -9.84, respectively, all P<0.01). (2) The protein expressions of FTO (0.69±0.14 vs.1.37±0.02) and Bcl-2 (0.50±0.12 vs.1.25±0.21) were significantly lower in I/R group than those of control group; While those of Bax (1.04±0.08 vs.0.57±0.06) and cleaved Caspase-3 (0.99±0.05 vs.0.36±0.07) were significantly higher ( t=8.10, 5.49, -8.22, and -12.09, respectively, all P<0.05). (3) Compared with the control group, the apoptosis rate of HK-2 cells in I/R group was significantly higher [(61.70±1.01)% vs.(0.16±0.10)%, t=63.80, P<0.01]. (4) Compared with the control group, the percentage of m 6A modification level in total RNA in I/R group was significantly higher [(3.13±0.21)% vs.(1.10±0.26)%, t=-10.61, P<0.01]. Conclusions:FTO-mediated RNA m 6A modification may affect renal IRI by regulating the apoptosis of HK-2 cells.

Objective To evaluate the effect of family doctor contracted service system in Shenzhen and explore the influencing factors for families to contract their family doctors. Methods A total of 480 residents were selected from 12 first-class community medical rehabilitation centers in Luohu, Futian and Nanshan districts of Shenzhen. The contents of the survey included the contracting status between the contracted and non-contracted residents, the utilization of four basic public health services, and the difference of satisfaction with community medical rehabilitation centers.Meanwhile, logistic regression was used to explore the important influencing factors of the contracted family doctors. Results The overall contracting rate of residents was 51.9% (248/478), and the lowest contracting rate was 23.1% (27/117) (P<0.05), found among patients under 40 years old.The utilization rate of contracted residents for growth and development examination, maternal health care as well as guidance of chronic disease prevention and treatment was higher than that of non-contracted residents ( P <0.05 ). fees and medical environment, service attitude,medical fees was higher than that of non-contracted residents(P<0.05).The satisfaction for convenience in medical environment, service attitude,medical fees and access to medical service was higher than that of non-contracted residents(P<0.05).The results of binary logistic regression analysis showed that age, education level, occupation ( retirement ) and chronic diseases were important factors affecting the signing of contracts.Conclusions The family doctors tend to be embraced by residents better than before. Family doctors contracted service system is conducive to improving the utilization rate of basic health services, acceptance rate of health services and satisfaction of residents. At the same time, the young population at large requires greater efforts before they can embrace the family doctor system.

Objective To study the mechanism of brain death-induced heart damage by observing the change patterns of morphological damage to the heart and related inflammatory factors after brain death and provide the experimental basis for heart transplantation by brain-dead donor.Methods The 30 rabbits were equally divided into two groups by the random digital table method:sham-operation group and brain death group.The rabbit brain death model was established in the brain death group,and the sham-operation group was given slow intracranial pressure.The rest treatments in the two groups were the same.At 2nd,6th and 8th h after operation,blood pressure,heart rate and respiratory rate were recorded.The damage of heart tissues was observed by HE staining.The plasma concentrations of IL-1,IL-6 and IL-8 were tested by ELISA.The expression of some inflammatory factors in heart issues was detected by RT-PCR and immunohistochemistry.Results At 8 h after brain death,there was no signifiant difference in blood pressure and heart rate between two groups (P＞0.05).The damage of heart issues in the brain death group was more serious than in the shamoperation group.With the prolongation of brain death,the plasma concentrations of IL-6 and IL-8 increased significantly in the brain death group (P＜0.05),but the concentration of IL-1β showed no siginificant difference between the two groups at 2 h after brain death (P＜0.05).Besides,the expression of HSP27 and HSP70 mRNA as well as the protein expression of ICAM and NF-κB was significantly increased in the brain death group as compared with that in the sham-operation group (P＜0.05).Conclusion With the prolongation of brain death time,the inflammatory factors in the heart tissues and plasma interleukin were increased,suggesting the inflammatory reaction occurs in donor heart under the condition of brain death,which influences the quality of donor in the heart transplantation.

OBJECTIVE: To screen new serum metabolic biomarkers for different drug resistance profiles of pulmonary tuberculosis (TB) and explore their mechanisms and functions. METHODS: We collected serum samples from TB patients with drug sensitivity (DS), monoresistance to isoniazid (MR-INH), monoresistance to rifampin (MR-RFP), multidrug resistance (MDR), and polyresistance (PR). The metabolites in the serum samples were extracted by oscillatory and deproteinization for LC-MS/MS analysis, and the results were normalized by Pareto-scaling method and analyzed using Metaboanalyst 4.0 software to identify the differential metabolites. The differential metabolites were characterized by function enrichment and co-expression analysis to explore their function and possible pathological mechanisms. RESULTS: Compared with the DS group, 286 abnormally expressed metabolites were identified in MR-INH group, 362 in MR-RPF group, 277 in MDR group and 1208 in PR group by LC-MS/MS analysis. Acetylagmatine ( < 0.05), aminopentol ( < 0.05), and tetracosanyl oleate ( < 0.05) in MR-INH group; Ala His Pro Thr ( < 0.001) and glycinoprenol-9 ( < 0.05) in MR-RFP group; trimethylamine ( < 0.05), penaresidin A ( < 0.05), and verazine ( < 0.05) in MDR group; and PIP (18:1(11Z)/ 18:3(6Z, 9Z, 12Z)) ( < 0.001), Pro Arg Trp Tyr ( < 0.001), N-methyldioctylamine ( < 0.001), and phytolaccoside E ( < 0.05) in PR group all showed significant differential expressions. Significant differential expressions of phthalic acid mono-2-ethylhexyl ester ( < 0.05) and eicosanoyl-EA ( < 0.05) were found in all the drug resistant groups as compared with DS group. CONCLUSIONS Acetylagmatine, aminopentol, tetracosanyl oleate, Ala His Pro Thr, glycinoprenol-9, trimethylamine, penaresidin A, verazine, PIP(18:1(11Z)/18:3(6Z, 9Z, 12Z)), Pro Arg Trp Tyr, N-methyldioctylamine, phytolaccoside E, phthalic acid mono-2-ethylhexyl ester, and eicosanoyl-EA are potentially new biomarkers that indicate monoresistance, multi-drug resistance and polyresistance of Mycobacterium tuberculosis. The combined use of these biomarkers potentially allows for assessment of drug resistance in TB and enhances the diagnostic sensitivity and specificity.
Biomarkers ; Chromatography, Liquid ; Humans ; Tandem Mass Spectrometry ; Tuberculosis, Multidrug-Resistant ; Tuberculosis, Pulmonary

OBJECTIVE：To overview and analyze the current evidence of systematic reviews of drug treatment for children with tic disorder （TD），and to provide evidence-based reference for clinical practice. METHODS ：Retrieved from Medline ，Embase， CBM，CNKI，VIP and Wanfang database ，and so on ，systematic reviews about the efficacy and safety of drug treatment for TD were collected. AMSTAR- 2 tools were used to evaluate the quality of included studies ；the intervention measures ，diagnosis standard，outcome index ，the types of included studies ，main conclusion ，control measures ，effect sizes of different drug treatment were also extracted and analyzed descriptively. RESULTS ：A total of 27 systematic reviews were included ，of which 5 were medium-quality，8 low-quality and 14 very-low-quality. Meta-analysis showed that although typical psychiatric drugs could control tic symptoms ，ADR were prominent and the safety was not good ；risperidone and aripiprazole as atypical psychiatric drugs had good efficacy and safety ，and the research evidence was relatively sufficient. Clonidine as α2 adrenergic agonist could effectively control tic symptoms with fewer ADR ，especially for patients with affention deficit and hyperactivity disorder （ADHD），and research evidence was sufficient. Tiapride could effectively improve tic symptoms ，and the overall tolerance was better. Tomoxetine and methylphenidate could significantly improve the tic symptoms of tic patients with ADHD. CONCLUSIONS ：In the clinical treatment of TD in children ，ADR induced by typical antipsychotics are prominent and the safety is not good ；risperidone， aripiprazole，clonidine and tiapride have good efficacy and safety. Clonidine ，tomoxetine and methylphenidate could significantly improve tic symptoms of TD patients with ADHD. But the quality of systematic review methodology for tomoxetine and methylp- henidate is not good and further improvement is needed.