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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.
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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.
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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.
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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.
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Objective To study the therapeutic effect of adjuvant chemotherapy and radiotherapy after operation in patients with mucoepidermoid carcinoma of parotid gland,and to screen the indicators ralated to the prognosis of tumor.Methods Eighty patients with mucoepidermoid carcinoma of parotid gland in First People′Hospital of Yibin of Sichuan Province from January 2005 to December 2009 were analysed retrospectively in our research.We studied the survival of patients who were treated wtih simple operation(30 cases)or postoperative adjuvant therapy(50 cases).Then we further analyzed the relationships between the prognosis of the patients and some variables (age,gender,smoking,alcohol drinking,lymph node metastasis,distant organ metastasis,treat-ment method,differentiation degree and T grading).Results Kaplan-Meier survival curves showed that patients with postoperative adjuvant therapy had longer PFS and OS than those without adjuvant therapy (94.4 months vs 69.3 months;114.9 months vs 96.7 months),with statistical significance (χ2 =11 .246,P =0.001 ;χ2 =15.803,P =0.001 ).COX univariate analysis showed that gender (χ2 =22.346,P =0.000),smoking (χ2 =7.891 ,P =0.041 ),lymph node metastasis (χ2 =12.371 ,P =0.005),distant organ metastasis (χ2 =9.81 3, P =0.002),treatment method (χ2 =25.261 ,P =0.000),differentiation degree (χ2 =4.361 ,P =0.006)and T grading (χ2 =5.336,P =0.01 4)were related to the PFS of patients.COX multivariate analysis showed that lymph node metastasis (χ2 =11 .003,RR =2.827,95%CI:1 .965-3.851 ,P =0.011 ),distant organ metastasis (χ2 =7.611 ,RR =0.472,95%CI:0.240-0.775,P =0.016),treatment method (χ2 =24.542,RR =5.390, 95%CI:3.585-9.602,P =0.000),degree of differentiation (χ2 =3.221 ,RR =2.1 1 8,95%CI:1 .845-4.719, P =0.009)and T grading (χ2 =4.336,RR =0.804,95%CI:0.681 -0.916,P =0.024)were related to the PFS of patients.COX univariate analysis showed that smoking (χ2 =4.551 ,P =0.008),alcohol drinking (χ2 =11 .742,P =0.048),lymph node metastasis (χ2 =14.886,P =0.009),distant organ metastasis (χ2 =6.71 3, P =0.005),treatment method (χ2 =22.411 ,P =0.000),degree of differentiation (χ2 =8.1 16,P =0.012)and T grading (χ2 =14.443,P =0.035)were related to the OS of patients.COX multivariate analysis showed that lymph node metastasis (χ2 =11 .711 ,RR =2.985,95%CI:1 .521 -3.999,P =0.005),distant organ metastasis (χ2 =5.390,RR =0.400,95%CI:0.201 -0.793,P =0.009),treatment method (χ2 =19.327,RR =5.086, 95%CI:3.241 -8.006,P =0.000),degree of differentiation (χ2 =7.084,RR =2.301 ,95%CI:1 .908-4.503, P =0.001 )and T grading (χ2 =1 3.229,RR =0.561 ,95%CI:0.348-0.867,P =0.040)were related to the OS of patients.Conclusion Adjuvant radiation and chemotherapy can obviously prolong the PFS and OS for the patients with mucoepidermoid carcinoma of parotid gland.Lymph node metastasis,distant organ metastasis,treat-ment method,differentiation degree and T grading can greatly influence the prognosis of patients with mucoepider-moid carcinoma of parotid gland,which can be used as independent prognostic indicators for the patients with mucoepidermoid carcinoma of parotid gland.
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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.
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Objective To analyze the epidemiological characteristics of mycoplasma pneumoniae (MP) infection among the out-patients and inpatients children in Shenzhen area during 2010-2012 and to explore the significance of the results of the laboratory routine tests in the diagnosis of MP infection .Methods The children patients with respiratory tract infection from 2010 to 2012 were selected and the MP infection and the non-MP infection were screened out .The epidemiological characteristics of gender ,age , etc .,among the children patients with MP infection during these 3 years were analyzed .The differences in the laboratory routine tests and high sensitivity C reactive protein (hsCRP) were compared between the MP infection and the non-MP infection .Results The positive detection rate of MP-DNA in males was slightly higher than that in females ,the difference had no statistical signifi-cance (P>0 .05);MP infection occurred in different age groups ,the positive detection rate of MP-DNA was lowest in the children patients aged <1 year old and highest in the children patients aged 3 - < 6 years (P< 0 .05);the routine laboratory tests and hsCRP level had no specificity in the diagnosis of MP infection .Conclusion The MP molecular epidemiology in Shenzhen area shows that MP infection has the seasonality ,the laboratory routine tests and hsCRP level can not be used as the basis of the MP la-boratory diagnosis .
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<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>Concentration 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.</p><p><b>RESULTS</b>The 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).</p><p><b>CONCLUSION</b>The 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.</p>