Objective To.investigate the accuracy and reliability of monocyte count of three methods.Methods The specimens classified by blood cell analyzer WBC monocytes ＜ 15％,15％-20％,＞ 20％ were taken randomly,draw 20 μ l after mixing,put in monocyte direct count dilution and count,and underwent routine smear,stained by Swiss-Gibbs and manual classified by microscope,and then compared the results of the three methods.Collected one copy each of the high,medium and low proportion of monocytes specimens,and counted 30 times by the three counting method.Results Blood cell analyzer in WBC monocytes 15％-20％,＞ 20％ had poor correlation with the results of the other two methods.Repeatability comparison of three methods,medium and low sample were better with method 1 and 2,high value sample was good with method 3.Conclusion The specimens of monocytes ≥ 15％,especially scatter plot is abnormal,must be confirmed by microscopic examination.

Objective To evaluate the effects of Pidomorph on serum immune cell level and T cell subsets in children with bronchial asthma.Methods 140 pediatric patients with bronchial asthma from January 2014 to October 2016 in our hospital were randomly divided into experimental group and control group according to the order of admission,70 cases in each groups.The control group was treated with conventional glucocorticoid drugs,and the experimental group was treated with pidotimod.The total effective rate of clinical treatment,the rate of rapid breathing in the flow rate,forced expiratory peak value,forced breathing vital capacity were compared between two groups three weeks after treatment,and the serum levels of immune cells(IgA,IgG,IgM)and T cell subsets(CD3+,CD4+,CD8+)were compared before and after treatment.Results The total effective rate was 92.86％(65/70)in the experimental group and 72.86％(51/70)in the control group,the total effective rate in the experimental group was significantly higher than that in the control group(P<0.05); Forced breathing speed in the speed,forced expiratory peak value,forced breathing vital capacity in the experimental group were better than the control group(P<0.05); After treatment,the levels of serum immune cells(IgA,IgG,IgM)in the experimental group were significantly higher than those in the control group(P<0.05); After treatment,the levels of T cell subsets(CD3+,CD4+,CD8+)in the experimental group were significantly higher than those in the control group(P<0.05).Conclusion Pidotimod is effective and safe in the treatment of bronchial asthma in children,it can effectively regulate T cell subsets and improve the immune function of children with asthma.

ObjectiveTo observe the effect of ginsenosides Rb1 on cerebral blood flow of rat models with cerebral ischemia/reperfusion (I/R) injury, which could provide a new theory of cerebral protective mechanism about ginsenosides Rb1.Methods Twenty-four rats were randomly divided into sham-operation group, model group, normal saline control group and ginsenosides Rb1 group, 6 rats in each group. The middle cerebral artery occlusion (MCAO) model was established by thread embolism method. At the end of I/R, in the rat of ginsenosides Rb1 group, ginsenosides Rb1 40 mg/kg was immediately intraperitoneally injected, while in the rat of normal saline control group, an equal volume of normal saline was injected intraperitoneally. After I/R for 24 hours, the cerebral local amount of blood flow was measured, the rats' behavior score was observed, and the volume of cerebral infarction was monitored by 2, 3, 5-triphenyl tetrazolium chloride (TTC) staining.Results The percentage of volume of cerebral infarction [(64.23±8.12)% vs. 0%] and behavior score [3.0 (2.0-4.0) vs. 0 (0-0),P< 0.05] in model group were significantly higher than those in sham-operation group, while the cerebral local amount of blood flow in model group was obviously lower than that in sham-operation group (mL/min: 125.75±57.65 vs. 225.01±78.25,P< 0.05); Compared with the model group and normal saline control group, the percentage of volume of cerebral infarction [(23.62±8.74)% vs. (64.23±8.12)%, 56.72±8.92] and behavior score [0.5 (0.0-2.0) vs. 3.0 (2.0-4.0), 3.5 (1.0-4.0)] in the ginsenosides Rb1 group were significantly lower, the cerebral local amount of blood flow was markedly increased in the ginsenosides Rb1 group (177.25±75.36 vs. 125.75±57.65, 132.65±58.65,P< 0.05).Conclusion Ginsenosides Rb1 can increase the cerebral blood flow in rats with cerebral I/R injury, which maybe one of the mechanisms of cerebral protection of Ginsenosides Rb1.

Objective To investigate the genetype distribution of mycoplasma pneumoniae(MP) by denaturing high-performance liquid chromatography(DHPLC).Methods A total of 300 cases nasopharyngeal aspirate were collected from our hospital.The MP genes of standard strains and clinical specimens isolates were amplified by PCR followed by DHPLC and genetype determination.Results A total of 110 cases were positive after 24 hours fermentation from 300 cases with pharyngeal swab.By the specific primers of MP-129,MP-FH standard strain and specimens,2 280 bp and 2 580 bp gene fragments were made out respectively.One hundred and ten strains of clinical isolates were detected by DHPLC.One hundred and seven strains of P1-Ⅰ were 1b subtype,3 were type P1-Ⅱ which were all 2a subtype.Conclusion The genetype of MP infection in children from our hospital is P1-Ⅰ,1b subtype by using DHPLC technology.

Through analyses of rare diseases management of several countries and regions , successful practice has been discussed .Legislation of rare diseases should be activated as soon as possible to release specific standard of rare diseases .Accelerating development of translational medicine is a feasible approach to improve the level of diag -nosis and treatment of rare diseases .Strengthening the orientation of management policy should be put into a more significant position to promote incentives of stakeholder in this area .Both enhancing the joint collaboration of differ-ent departments and perfecting medical insurance system could be forceful accelerants to create and optimize the management system of rare diseases in China .

Objective To discuss the effectiveness of anterolateral thigh propeller flaps for treatment of inguinal skin and soft tissue defects.Methods From June,2009 to October,2014,12 patients with inguinal skin and soft tissue defects were treated with anterolateral thigh propeller flaps pedicled with perforator of descending branch of lateral circumflex femoral artery.Of them there were 8 males and 4 females,aged from 22 to 51 years.The left side was involved in 3 cases and right side 9 cases.Defects were caused by traffic accident injury in 5 cases,crash injury of heavy object in 7 cases.There were mere skin and soft tissue in 2 cases,combined with bone fractures,nerves,vessels and muscles injury in 10 cases.The area of defects ranged from 9.0 cm×5.0 cm to 22.0 cm×9.0 cm.During operation,anterolateral thigh propeller flaps ranging from 11.0 cm×7.0 cm to 24.0 cm×1 1.0 cm were used to repair the wounds.Results All flaps and skingrafts survived after operation and the wounds obtained primary healing.After 8-24 months follow-up,all flaps were characterized by soft texture,good color,and satisfactory appearance.According to the Britain's Medical Research Council at 8 months after operation,the sensation of the flaps were recovered to S2 ～ S3+,No obvious scar contracture and chromatosis were observed at donor site.Conclusion Anterolateral thigh propeller flaps pedicled with artery descending branch of lateral circumflex femoral perforator is an ideal choice for the reconstruction for inguinal skin and soft tissue defects.

Objective To examine the association of urinary neutrophil gelatinase-associated lipocalin(NGAL) and interleukin 18(IL-18) with acute kidney injury (AKI) in patients after cardiac surgery. Methods Thirty-three patients undergone cardiac surgery were divided into AKI group and non-AKI group according to the AKI criteria. The Scr, urinary NGAL and IL-18 were measured at different time points. Results Nine of 33 patients (27.27%)developed postoperative AKI, and Scr concentration in AKI group reached its peak within 12-48 hours after cardiac surgery. Urinary concentrations of NGAL and IL-18 at 2 h and 4 h after cardiac surgery were significantly higher than those before operation in AKI patients (P<0.01). The urinary concentrations of NGAL at each time point and that of IL-18 at 2 h and 4 h after cardiac surgery in AKI patients were significantly higher than those in non-AKI patients. After correction by urinary creatinine, the differences of NGAL/Ucr and IL-18/Ucr ratios were still significant (P< 0.01). For concentrations of urinary NGAL, IL-18 and ratios of NGAL/Ucr, IL-18/Ucr at 2 h after surgery, sensitivities and specificities were good with cutoff values at 250 μg/L, 250 μg/mmol and 1800 ng/L, 1800 ng/mmol, respectively. Urinary concentration of NGAL at 2 h after cardiac surgery was positively correlated with Scr at 12 h postoperation in AKI group (r=0.638, P<0.05).Conclusions The incidence of AKI in patients after cardiac surgery is quite high. Urinary concentrations of NGAL, IL-18 and ratios of NGAL/Ucr, IL-18/Ucr at 2 h after cardiac surgery are the early diagnostic markers for AKI, among which urinary NGAL/Ucr is the most sensitive one.

Objective To investigate the value of percutaneous transhepatic cholangioscopy (PTCS) for diagnosis and treatment of biliary cast after liver transplantation. Methods Data of 11 patients with biliary cast after liver transplantation, who underwent PTCS from April 2008 to November 2010, were retrospectively analyzed. Results In 11 patients , one had biliary cast in common bile duct, 3 in right intra-hepatic bile duct, 4 in left intra-hepatic bile duct, and 3 distributed in intra- and extra-hepatic bile ducts. A total of 68 times of PTCS were performed in 11 patients, achieving significant decrease in levels of serum transaminase and bilirubin in 10. Occasional fever occurred in 1 patient after closure of drainage tube,which was managed by replacement with a thinner one. There were no severe complications such as biliary fistula or uncontrollable bleeding. Partial rupture of fistula occurred in 1 case. All patients were followed up for 10-30 months and were all in good condition except one patient died from other disease during the followup. Conclusion PTCS is a safe, effective and applicable method to treat the biliary cast after liver transplantation.

Objective : To analyze the epidemiological characteristics of food-borne diseases in Jinshan District, Shanghai from 2014 to 2020, so as to provide the evidence for formulating the public health strategy for food-borne diseases control. Methods : The medical records of patients with food-borne diseases were collected from 16 monitoring hospitals in Jinshan District from 2014 to 2020, and the basic information, clinical symptoms, history of suspicious dietary exposure and disease diagnosis were extracted. The crowd distribution, temporal distribution, spatial distribution, history of suspected dietary exposure and etiological characteristics of patients with food-borne diseases were descriptively analyzed. Results : A total of 1 060 cases with food-borne diseases were reported in Jinshan District from 2014 to 2020, including 1 057 cases with infectious diseases ( 99.72% ) and 3 poisoning cases ( 0.28% ). The male/female ratio of the cases was 0.94∶1, and 47.55% ( 504 cases ) were at ages of 15 to 44 years. Working ( 402 cases, 37.92% ) and farming ( 218 cases, 20.57% ) were predominant occupations, and the detection of food-borne diseases was concentrated between May and October, with two peaks seen in August and May. The suspicious food exposure was predominantly meat and meat products (215 cases, 20.28%), and the suspicious food exposure place was predominantly at home ( 363 cases, 34.25% ). In addition, the positive rate of food-borne infection was 23.03% in 712 samples, including 123 samples with Vibrio parahaemolyticus infections ( 17.83% positive rate ), and 26 samples with Salmonella infections ( 3.65% positive rate ). Conclusion Food-borne diseases were highly prevalent in summer in Jinshan District from 2014 to 2020, and infectious cases were predominant. Young people, workers and farmers are at high risk of food-borne diseases, and Vibrio parahaemolyticus and Salmonella were predominant pathogens.

To obtain and analyze the sequence of the nucleocapsid gene from bovine coronavirus, and to produce the fusion protein of the N gene in E.coli in order to use this recombinant protein for the study of bovine coronavirus. The N gene of BCV-DQ strain was amplified by RT-PCR, in which the primers were designed on the basis of N gene sequence of BCV-Mebus strain. The PCR products of 1 347 bp in length were cloned and sequenced, and then inserted into the prokaryotic vector pET30a. The recombinant plasmids were then transformed into Escherichia coli BL21 and identified by SDS-PAGE and Western blot assay. ELISA assay was optimized of N protein as the coating antigen to detect the viruses in the clinical samples. In comparison with 6 BCV strains in GenBank, the sequence identity was proved to be more than 98.3%. Result in SDS-PAGE showed that the fusion protein had a molecular weight of 60 ku, and could be specifically recognized by mouse serum against BCV. The indirect ELISA was used to test 256 serum samples collected from Heilongjiang province and 65.23% samples were positive. On testing field samples, an overall agreement of 95.31% was generated between the the neutralization test of viruses (VN) and indirect ELISA. It is apparent that the N gene was highly conservative and is expressed in E. coli in high level,also the prokaryotic expression products of this gene show a fine reactiongenicity in immune responses. It was also suggested that the N protein may be a useful antigen for sero-diagnosis and epidemiological investigation of BCV.