Objective This study was to analysis the epidemiologic characteristics of hand-foot-month (HFMD) in Shenzhen during 2015-2016,to provide reference for the prevention and treatment of HFMD.Methods 7 758 statistical data from Shenzhen children's hospital clinical lab during 2015-2016 was included.We used real time fluorescent reverse transcription polymerase chain reaction (RT-PCR) to detect enterovirus general (EV),enterovirus 71 (EV71) and coxsackievirus A group 16 (CoxA16),and analyzed the age,sex and epidemic time of the patients.Results In 2015 and 2016,the positive rate of EV was 67.19％ (2679/3987) and 52.61％ (1 984/3 771) respectively,with statistically significant difference (x2 =71.84,P ＜ 0.05).The radio of male to female children was 1.91∶1 (1 757/922) and 1.83∶1 (l 283/701) in 2015 and 2016,with statistically significant difference (P ＜ 0.05).The age of the children was ＜ 5 years old,accounting for more than 90％ of the total number of patients.April to July and September were the two peaks of HFMD.The enterovirus of hand foot and mouth disease in Shenzhen was dominated by other enteroviruses (more than 82％ of the total number of patients).With the increase of age,the proportion of EV71 in children with severe hand and foot was increasing,and the proportion of other enteroviruses was decreasing.Conclusions Vaccination is one of the important measures of HFMD control.It's beneficial for the diagnosis and treatment of HFMD to collected epidemiologic characteristics data about HFMD in Shenzhen.

Objective To investigate the genotype and mutation frequency of thalassemia in child patients of Shenzhen region so as to provide evidences for the gene diagnosis and genetic counseling of thalassemia.Methods A total of 1 206 child patients suspected with thalassemia were retrospectively analyzed.The gene deletion of α-thalassemia was detected by Gap-PCR.The point mutations of α-thalassemia and β-thalassemia were determined by reverse dot blot(RDB)-PCR.The specimens suspected with HKαα and rare gene mutations were determined with nested PCR and gene sequencing,respectively.Results The detection rate of thalassemia was 76.9％ (927/ 1 206).Among them,α-thalassemia accounted for 40.5％ (489/1 206),and--SEA/αα was the most common gene mutation(75.1％);β-thalassemia accounted for 33.7％ (406/1 206),and the main IVS-2-654 (C→T) and CDM1-42 (-TCTT) heterozygous mutations accounted for 35％ and 32.5％,respectively.In addition,there were 32(2.7％) β-thalassemia patients with α-thalassemia mutation,1 patient with HKαα/ααQS,1 α-thalassemia patient with CD61 (AAG→TAG)/--SEA and 1 β-thalassemia patient with CD5 (CCT→C).Conclusion The are complicated gene mutation types and rare gene mutations of thalassemia in child patients of Shenzhen region.

Since the reform of the supply side has been put forward by General Secretary Xi Jinping,the research and practice of the health industry has been strengthened for the reform of the supply side,and the beneficial exploration has been carried out on the supply side structure reform.The various forms of medical alliance are the feasible way to solve the problem of supply side.Through such a reform,it can make full use of medical resources to meet the needs of the people's medical treatment.The construction of medical alliance practice in Beijing Chaoyang Hospital is analyzed,and on this basis,proposals of the medical alliance construction are put forward.

Objective To investigate the effects of tongue cancer resistance-associated protein 1 (TCRP1) in proliferation of chronic myeloid leukemia cells (CML),and explore the new thoughts of pathogenesis of CML.Methods The expression of TCRP1 was detected in the peripheral blood mononuclear cells (PBMC) of CML with real-time quantitative polymerase chain reaction (PCR) and Western blot.After the expression of TCRP1 was interfered in K562 cells,the proliferation of cells was detected by 3-(4,5-dimenthylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay and soft agar colony forming assay,and the expression of protein kinase B (AKT) and its phosphorylation were tested by Western blot.Results In PBMC of CML patients,the mRNA and protein levels of TCRP1 were significantly higher than those of normal controls.The results of MTS assay and soft agar colony forming assay showed that the proliferation of K562 cells was significantly decreased after the expression of TCRP1 was interfered.After knockdown of TCRP1 in K562 cells,the phosphorylation of AKT was significantly decreased while the expression of total AKT did not change.Conclusions The expression of TCRP1 was increased in CML cells.High expression of TCRP1 might contribute to proliferation of K562 cells via the phosphorylation of AKT.

Objective To establish the pipeline and evaluate the feasibility of high-throughput sequencing method used in the detection of severe pneumonia pathogens.Methods Clinical control study was used.Bronchi alveolar lavage fluids (BALF) samples from 76 patients with severe pneumonia (treatment group) and 18 patients with tracheal malacia (control group) and without clinical detected pathogens were collected during March 2015 to December 2016 in Shenzhen Children′s Hospital.The pathogens in the samples were detected using clinical tests and high-throughput sequencing respectively.The results of high-throughput sequencing were confirmed by real-time quantitative PCR and the high-throughput sequencing method used in the detection of severe pneumonia pathogens was evaluated.The χ2 test was used to analyze the correlation of detection rate between the high-throughput sequencing group and the non high-throughput sequencing group.Results The pipeline and method of high-throughput sequencing used in the severe pneumonia pathogens detection was established.The pipeline included sample collection, DNA extraction, library construction, sequencing, and bioinformatic analysis.In 76 cases of patients with severe pneumonia, the results of high-throughput sequencing in 66 cases of bronchoalveolar lavage fluid specimens were positive.The sensitivity was 86.84%, which was significantly higher than the total sensitivity of traditional clinical detection methods including bacterial culture, immunofluorescence and quantitative PCR(68.42%,52/76),χ2=7.426,P<0.001.A total of 13 pathogens were detected in 66 positive samples of high-throughput sequencing, including Mycoplasma pneumoniae, Streptococcus pneumoniae, Haemophilus influenzae and adenovirus, etc.Nine kinds of pathogens were detected in these samples through non-high-throughput sequencing.In the experimental group, the results obtained by clinical test and high-throughput (80.26%) were entirely consistent in 61 samples and not completely consistent in 15 samples (19.74%) specimens.These inconsistent results were mainly concentrated in the detection of adenovirus, Streptococcus pneumoniae and Haemophilus influenzae through high-throughput sequencing, whereas clinical cultures and immunofluorescence methods were not able to detect these pathogens.PCR validation showed that there was no significant difference between the results of high-throughput sequencing and the PCR tests (χ2=0.517,P=0.472), and the difference between the results of high-throughput sequencing and traditional clinical detection methods was statistically significant (χ2=11.671,P<0.001).Conclusion The method for the detection of severe pneumonia pathogens based on high-throughput sequencing is highly sensitive and can detect unknown pathogens, which is superior to those used in traditional clinical detection.

Objective To perform a risk factor analysis of central retinal vein occlusion (CRVO) and branch retinal vein occlusion (BRVO),and compare the difference in risk factors between CRVO and BRVO.Methods Retrospective observational casecontrol study included 46 CRVO patients,33 BRVO patients and 79 control subjects with senile cataract or refractive error,the risk factors and blood lipid spectrum analysis were performed and compared.Results Multivariate linear regression analysis showed that higher serum levels of homocysteine (P ＜ 0.000 1),total cholesterol (P =0.003 0),lipoprotein (a) (P =0.027 0),hypertension (P =0.022 0) and shorter axial length (P ＜0.000 1) were significantly correlated with CRVO.BRVO was associated with higher serum levels of homocysteine (P ＜0.000 1),total cholesterol (P =0.008 0),hypertension (P =0.002 0),body mass index (P =0.004 0) and shorter axial length (P =0.001 0).There was no significant difference in risk factors between CRVO and BRVO patients on multivariate analysis.Conclusion Multiple systemic (hyperlipidemia,hypertension and hyperhomocystinemia) and ocular (shorter axial length) risk factors are associated with both CRVO and BRVO,but these risk factors are not different between CRVO and BRVO.

Objective To investigate the effect of racecadotril granules combined with montmorillonite on infantile rotavirus enteritis and myocardium enzymogram.Methods Retrospective a total of 275 infants and young children with rotavirus enteritis treated in Seventh Hospital of Ningbo from September 2014 to January 2016,were divided into the control group and the combined treatment group according to the treatment method.The combined treatment group were treated with montmorillonite powder and racecadotril granules, the control group were treated with montmorillonite powder.The negative rate of stool were observed three days after treatment,compared the symptom relief time,the level of inflammatory cytokines and myocardial enzyme spectrum between two groups before and after treatment .Results The stool negative rate of combined treatment group ( 79.31%) higher than the control group(67.69%) after three days of treatment;the levels of IL-18,IL-6 and hs-CRP in the combined treatment group were lower than those in the control group(P<0.05);the antiemetic time,dehydration correction time and recovery time of stool were shorter in combination therapy group than those in control group(P<0.05);there was no difference in the levels of myocardial enzyme between the two groups before treatment.The level of CK-MB,LDH,CK and AST in the two groups after treatment were lower than before treatment,and the level of CK and CK-MB in the combined treatment group was lower than the control group ( P<0.05 ) .Conclusion Racecadotril granules combined with montmorillonite powder has a good therapeutic effect on infantile rotavirus enteritis can significantly improve clinical symptoms, reduce the levels of inflammatory cytokines in patients.

Objective To investigate the clinical effect of combined treatment of children allergic purpura with traditional Chinese and western medicine and preventive effect on renal damage ,and provide the basis for clinical therapy.Methods According to the different treatment methods ,96 children with allergic purpura were divived into the treatment group and the control group ,each group in 48 cases.The treatment group treated by combined western and Chinese medicine ,the control group was treated with western medicine ,the symptoms disappear time observation of treatment of the two groups of children,urinary N -acetyl -D beta glucosidase(NAG),urine albumin(mALB) changes,compared two groups of children with renal damage occurrence time and the incidence .Results The treat-ment group purpura disappearance time ,joint pain and swelling disappeared time ,abdominal pain disappeared time were (5.15 ±2.26)d,(3.12 ±0.81) d,(2.06 ±0.16) d,the control group respectively were (8.03 ±3.08) d, (5.13 ±1.09)d,(4.76 ±1.04)d.The difference between the two groups had statistical significance (t =2.01, 11.03,24.54,all P<0.05);After three months of treatment,the treatment group abnormal urine rate,NAG,mALB were 16.7%,(5.46 ±3.20) u/L,(7.24 ±7.62) mg/L,the control group were 37.5%,(7.58 ±4.32) u/L, (19.40 ±3.48)mg/L,the difference between the two groups was statistically significant (χ2 =6.67,t=2.72,10.13, all P<0.05).The treatment of renal damage occurred time [(36.4 ±12.4) d] later than that of the control group [(22.1 ±10.6)d](t=6.08,P<0.05),the incidence of renal damage in treatment group (16.7%) was lower than that in the control group (41.2%) (χ2 =7.26,P<0.05).Conclusion Compared with simple western medicine treatment,the treatment of children allergic purpura with combination of TCM and western medicine can reduce the symptoms disappeared time,reduce the content of NAG,mALB,reduce the incidence of renal damage ,and prevent kidney damage .

OBJECTIVETo investigate the protective mechanism of stearoyl-CoA desaturase 1 (SCD1) over-expression against the pro-apoptotic affects of palmitic acid on hepatocytes using the rat BRL cell line.

METHODSConcentration effect curves were generated using the trypan blue exclusion test to assess the death rate of BRL cells upon exposure to a dilution series of palmitic acid. The multiplicity of infection (MOI) of a lentiviral expression vector, pGC-FU-GFP, was determined for the BRL cells. Unmanipulated BRL cells were divided into two groups: the non-palmitate groups were composed of ordinary cultured cells (CON) alone, infected with lentivirus empty expression vector (negative control, NC), and infected with lentivirus overexpressing SCD1 (SCD1-LV); the palmitate groups were composed of ordinary cultured cells plus palmitate (CON+) alone, infected with lentivirus empty expression vector plus palmitate (NC+), and infected with lentivirus overexpressing SCD1 plus palmitate (SCD1-LV+). SCD1 mRNA expression was detected by real-time PCR. Propidium iodide (PI) single-staining was used to detect apoptosis and assess the cell cycle. Inter-group differences were analyzed statistically.

RESULTSThe death rate of BRL cells increased significantly after 72 h of exposure to 400 mumol/L palmitate (P less than 0.01). The MOI of pGC-FU-GFP in BRL cells was 20. The expression of SCD1 was significantly higher in the SCD1-LV and SCD1-LV+ groups than in the respective controls (vs. CON: F = 289, P less than 0.01; vs. CON+: F = 1522, P less than 0.01). Palmitate exposure led to decreased expression of SCD1 (CON+ vs. CON, F = 22, P less than 0.05 and NC+ vs. NC: F = 34, P less than 0.05). The ratio of S stage cells was similar in all non-palmitate groups (CON, NC and SCD1-LV, P = 0.137). However, there was a significant apoptotic peak and lower ratio of S stage cells in the control palmitate groups (CON+ and NC+) and the activity of cell proliferation was decreased as well. The ratio of apoptotic cells was decreased significantly in the SCD1-LV+ group compared to the CON+ group (P less than 0.01).

CONCLUSIONThe expression of SCD1 and its desaturation activity increased in BRL cells upon infection with the pGC-FU-SCD1-GFP lentiviral vector, suggesting that SCD1 over-expression can decrease palmitic acid-induced toxicity and apoptosis in hepatocytes.

Animals ; Apoptosis ; Cell Line ; Genetic Vectors ; Hepatocytes ; metabolism ; Lentivirus ; genetics ; Palmitic Acid ; toxicity ; Rats ; Stearoyl-CoA Desaturase ; metabolism

Objective To evaluate the performance of ACL TOP coagulation analyzer system in the laboratory of children′s hospital. Methods According to the documents of CLSI, the analytic characteristics including precision, accuracy, linearity, interference and carryover rate were examined; specimens from healthy children were collected and assayed to determine the reference range of Prothrombin Time (PT), Activated Partial Thromboplastin Time (APTT), Thrombin Time (TT), Fibrinogen (FIB) and D-Dimer (D-D) from children on ACL TOP. Results The within-run and between-day coefficient of variability (CV) were within an acceptable range; The accuracy deviation of PT , APTT and FIB were less than 1/2 allowed total errors; The results of determination of FIB linearity test were correlated with the results of calculation: Y = 1.002 1X-0.122, R2 =0.998 2; The extent of influence of low to middle grade of jaundice , fat and hemolysis on each test were all less than 1/2 allowed total error; The carryover rates were lower than 1.81% and within an acceptable range; The reference range of PT, APTT, TT and FIB were PT (9.1-13.1 s), APTT (24.9-42.1 s), TT (12.6-21.1 s), FIB (1.924-4.011 g/L). Conclusion The ACL TOP coagulation analyzer has good repeatability, stability, linearity and capability of anti-interference and anti-carryover.