Objective To establish the reference ranges of peripheral blood T lymphocyte subsets among 333 healthy adults in Shanghai,to analyze and discuss the change characteristics through the establishment of 95%confidence interval,and to provide the reference for immune status analysis and clinical diagnosis and treatment. Methods Multiparameter three-color immunofluorescence was used to stain the peripheral blood T lymphocytes,and BD FACSCalibur flow cytometry was used to acquire and analyze the percentages and absolute counts of T lymphocyte subsets.Results Among females,the CD3 +percentage,CD4 +/CD8 +ratio,CD3 +CD4 +percentage and CD4 +CD45RA+percentage (72.6% ±8.1%),1 .57(0.74 -4.70),(44.8% ±8.0%)and (1 .71% ±6.3%)were respectively higher than those among males (70.9%±8.5%),1 .43(0.51 -4.1 2),(40.6%±9.3%)and (1 5.1%±6.8%).The lymphocyte absolute count and CD8 +CD28 +absolute count in males (2 069.8 ±474.1 )cells/μL and (237.5 ±95.7) cells/μL were higher than those in females (1 932.8 ±469.7 ) cells/μL and (21 8.1 ± 1 1 1 .1 )cells/μL.The percentage of T lymphocyte subsets increased with the increase of age.There were statistical significances in >50 year-old group with the other groups.The absolute count of 333 healthy adults for peripheral blood T lymphocyte was (1 069.1 -2 935.6)cells/μL.The percentages of T lymphocyte subsets were:CD3 +55.3% -87.6%,CD3 +CD4 +25.2% -57.6%,CD3 +CD8 +1 5.0% -42.1%,CD4 +CD25 +2.5% -1 0.5%,CD4 +CD45RA+5.0%-27.2 % and CD8 +CD28 +4%-1 9.3%,and CD4 +/CD8 +ratio was 0.7 -2.9.Conclusions There are differences for the percentage distribution of T lymphocyte subsets among healthy adults between sex and age. Since these differences can be ignored,the reference ranges of peripheral blood T lymphocyte subsets among healthy adults could be established in Shanghai.

Objective To detect the expression of soluble B7H3in serum of lung cancer patients,and to explore the clinical significance of soluble B7H3in diagnosis and treatment.Methods 196specimens out of the corresponding patients who had been diagnosed with lung cancer by Longhua Hospital Shanghai University of TCM were collected.50patients with other pulmonary diseases(OPD)were chosen as OPD group.60specimens of healthy people were collected as normal group.All the sB7H3levels of the specimens were detected by ELISA and then the expression characteristic of sB7H3could be analyzed.Results The sB7H3levels in lung cancer patients were significantly higher than that of OPD group and normal group(P<0.01).There was no significance between OPD group and the normal group(P=0.139).No significant difference of sB7H3levels was observed in patieits of different sex,age,histological subtype or tumor stage(P=0.503,0.133,0.810,0.074).The sB7H3levels of lung cancer patients with operation were significantly higher than those without operation(P=0.014).Conclusion sB7H3is a valuable biomarker for lung cancer.And it suggests a clinical significance in the anriliary diagnosis of lung cancer.

Objective To explore and evaluate the clinical value of the detection of C-reaction protein in children's respiratory infections.Methods To retrospectively analyze the infection situation of 7 respiratory viruses [parainfluenza Ⅰ(PIVⅠ),parainfluenza Ⅱ(PIVⅡ),parainfluenza Ⅲ(PIVⅢ)],respiratory syncytin virus(RSV),influenza virus A(IVA),influenza virus B(IVB) and adenovirus(ADV)) among 779 children with lower respiratory tract infections in Longhua Hospital.And then explore these results combined with their serum levels of CRP.Results Among 779 samples,189 samples were shown to be viral positive which including 117 cases of single virus infection(61.90%) and 72 cases of mixed infection(38.10%).The total positive ratio was 24.26%.The serum level of C-reaction protein was shown to be positive in 44.44% children with more than two virus infection while that to be 24.79% as single virus infection,the difference was statistically significant(P<0.05).Conclusion The children have more possibility of bacterial infections when they have mixed respiratory viral infections because their immune functions are inhibited.So serum level of CRP might be used as one of the important monitoring indicators.

Objective To investigate the coorelation of pathological diagnosis,TNM stage and the levels of tumor markers with different scenarios of traditional Chinese medicine syndrome in patients with lung cancer.Methods A total of 196 patients had been diagnosed with lung cancer.The correlation analysis of pathological diagnosis [non-small cell lung cancer(squamous carcinoma,adenocarcinoma,large cell carcinoma and mixed cell carcinoma) and small cell lung cancer],TNM stage and tumor markers[cytokerantin-19-fragment(CYFRA21-1),carcinoembryonic antigen(CEA),carbohydrate antigen 125(CA125),pro-gastrin-releasing peptide(ProGRP) and squamous cell carcinoma antigen(SCC Ag)] with different scenarios of traditional Chinese medicine syndrome in patients with lung cancer was performed.Results Most of the traditional Chinese medicine syndromes in lung cancer patients were lung and spleen deficiency syndrome(119 cases,60.71%),Qi and Yin deficiency syndrome(76 cases,38.7%) and spleen and kidney deficiency syndrome(1 case,0.52%).However,the distribution of the traditional Chinese medicine syndrome among the different pathological diagnosis in lung cancer patients showed no significant difference(P=0.325).Lung and spleen deficiency syndrome was concentrated in TNM stage Ⅰ-Ⅲ patients,while Qi and Yin deficiency syndrome was concentrated in TNM stage Ⅳ patients.The CYFRA21-1 levels in Qi and Yin deficiency syndrome patients were higher than those in lung and spleen deficiency syndrome patients(P=0.017).The differences of CEA,CA125,ProGRP and SCC Ag levels were not significant(P0.05).Conclusions Most of the lung cancer patients are deficiency syndrome.It indicates that advanced stage lung cancer patients would be in unstable conditions and have bad prognosis when Qi and Yin deficiency.CYFRA21-1 and other tumor markers could be the microscopic evidence to traditional Chinese medicine syndrome diagnosis.

Objective To analyze the infection situation of 7 common viruses [parainfluenza virus Ⅰ(PIVⅠ),parainfluenza virus Ⅱ(PIVⅡ),parainfluenza virus Ⅲ(PIVⅢ),respiratory syncytial virus(RSV),influenza virus A(IVA),influenza virus B(IVB) and adenovirus(ADV) ]among children with acute lower respiratory tract infection.Methods Nasopharyngeal secretion was collected from 2 425 hospitalized children with acute respiratory tract infection,and 7 respiratory viruses were detected to analyze the clinical epidemiological characteristics by age group [1 526 cases of infants(≤3 years old),650 cases of preschools(4-7 years old) and 250 cases of children(≥8 years old) ],virus distribution and seasons.Results Among 2 425 children,552 cases were shown to be positive for 7 viruses which included 360 cases of single virus infection(65.22%) and 192 cases of mixed infection(34.78%).The total positive rate was 22.76%.In different age groups,the number of infected infants were 377 cases(68.29%),including 242 cases of single virus infection(43.84%) and 135 cases of mixed infection(24.46%).Among the 7 viruses,the infections were mainly related to PIV Ⅲ infection with 196 cases(35.51%),followed by RSV infection [183 cases(33.15%) ].In 4 seasons,the positive rates were 17.79%(spring),19.73%(summer),22.37%(autumn) and 31.01%(winter),respectively.The positive rate in winter was the highest.Conclusions PIVⅢ is the major virus in children with respiratory tract infection.The respiratory tract viral infection rate in infants(≤3 years old) is the highest.The respiratory viruses are most prevalent in winter.

Objective To analyze the characteristics of precore/core gene mutations in hepatitis B virus carriers with coexistence of HBsAg and HBsAb(double positivity).Methods Specimens collected from 18 cases with double positivity were detected for the mutation of precore/core gene and S gene.Results 7 cases with precore/core gene mutations were demonstrated,who were with more S gene mutations and higher mutation rate than those cases without precore/core mutation(P<0.05).4 of the cases with precore/core gene mutations were positive with nt1896 mutation in precore gene,of which 1 case was HBeAg-negative and 3 cases HBeAg-positive.Conclusion Hepatitis B virus carriers with double positivity might be not only with more S gene mutations,but also with more precore/core gene mutations.nt1896 mutation in hepatitis B virus carriers with double positivity could be more common in HBeAg-positive patients than HBeAg-negative patients,which might be associated with the complexity of HBV isolates.

Retrospective analysis on the results of antinuclear antibody test,to investigate the correlation between indirect immunofluorescence assay for screening ANA and line immunoassay for specific ANA,752 specimens were tested by IIF-ANA and LIA-ANAs.Among 752 cases,the positive rate of the two methods were 27.26%(IIF) and 27.66%(LIA),and two methods showed no statistically difference(P=0.875,kappa=0.463).ANA-positive rate of men and women showed significant difference(15.5% vs.32.71%,P=0.000).The detection rate of IIF for specific autoantibodies such as anti-Jo-1,anti-PM-Scl and anti-Scl-70 is low.IIF+/LIA+ patients had high-titer≥ 1∶1000,accounting for 64.3%,while IIF+/LIA-patients had low-titer 1∶100,accounting for 50.5%.IIF can be used for screening the antinuclear antibodies,then LIA for confirmation.For those IIF+/LIA-patients: if they have obvious clinical symptoms of AID but IIF test is negative,LIA test should be performed for confirmation;for those IIF+/LIA-patients with high titer,especially in those patients with subclinical symptoms,clinical attention should be given,and regular follow-up is recommended.

To evaluate the correlation of T cell subsets,serum immunoglobulin and complement levels with different kinds of syndromes of ZhongYi of post hepatitis liver cirrhosis,84 patients have been diagnosed as four kinds of syndrome of Zhong Yi including 40 patients with syndrome of accumulated dampness-heat(group①),20 patients with syndrome of blockade of dampness due to qistagnation(group②),12 patients with syndrome of Yin deficiency of liver and kidney(group③),11 patients with syndrome of Yang deficiency of spleen and kidney(group④).T-lymphocyte subsets(by flowcytometry) and serum immunoglobulin levels were measured for all the patients.Compared with control group,the IgA、IgG levels were increased significantly in all group(P0.05,P0.01).CD8+ T cells and NK amounts were decreased(P0.05)in group①.CD8+T cells were decreased significantly(P0.01),and CD4+ T cells were decreased(P0.05)in group②.CD4+/CD8+ ratio was significantly increased in group ② than in any other groups.The different kinds of syndrome of ZhongYi were related to status of immunity,and the syndrome of ZhongYi changed with the status of immunity.CD4+,CD8+,CD4+/CD8+ ratio,immunoglobulin levels determinations may help to define syndrome of ZhongYi and used to clinical diagnosis and treatment.

Objective To study the double-peak phenomenon on the direct sequencing chromatograms from HBsAg+/HBsAb+patients in previous study,observe the change of nucleotide mutations on after cloning and analyze its homology.Methods HBsAg+/HBsAb+patients with the double-peak phenomenon were tested as an experimental group (n=3),and HBsAg+/HBsAb- patients with the double-peak phenomenon were served as a control group (n=4).The S gene of HBV DNA was amplited,cloned and sequenced.Results The sequencing results were not same.Furthermore,the rate of mutation of the experimental group was signficantly higher than contrd group.In addition,there were no statistically significant difference in homology among both groups.Conclusion The HBV genome of the experimental group is more complex than that of the control group.There are some kinds of different HBV strains in one patient,which is the cause of the double-peak phenomenon in direct sequencing.However,the different nucleotide mutations between HBV strains were too little to represent a different genotype or serotype,which is called a quasispecies.

Objective To investigate the relationship between hepatitis B e antigen(HBeAg) and alanine aminotransferase(ALT) levels among chronic hepatitis B(CHB) patients with different loads of hepatitis B virus(HBV) DNA,and to provide the reference for clinical treatment.Methods A total of 214 CHB patients with ALT within normal range and 200 healthy subjects with HBsAg-negtive and normal ALT levels were enrolled.The ALT levels′ differences were compared among HBeAg(+) group of CHB patients,HBeAg(-) group of CHB patients and healthy subjects.The 214 cases of CHB patients with normal ALT levels were classified into 3 groups according to HBV DNA load(GroupⅠ: HBV DNA103 IU/mL,Group Ⅱ:103 IU/mL≤HBV DNA104 IU/mL and Group Ⅲ: HBV DNA≥104 IU/mL).ALT levels and the positive rate of HBeAg of these groups were analyzed.Results Among the 214 CHB patients,the ALT levels of HBeAg(+) group and HBeAg(-) group were higher than that of the healthy group(P0.01),and the ALT levels of HBeAg(+) group were higher than those of HBeAg(-) group(P0.05).The positive rate of HBeAg of Group Ⅰwas significantly lower than those of Group Ⅱ and Group Ⅲ(P0.05).The ALT levels of Group Ⅲ were significantly higher than those of Group Ⅰ and Group Ⅱ(P0.05).Conclusions The ALT level is at the upper limit of normal range,and the positive rate of HBeAg is rather high in CHB patients with ALT within the normal range and HBV DNA≥104 IU/mL,which suggests that more attention should be paid for the patients in the clinical course of treatment.

Objective To analyze relationship between genotypes and mutations in S gene sequence of chronic hepatitis B patients with positive results for both HBsAg and HBsAb tests(HBsAg+/HBsAb+). Methods Specimens collected from 43 cases of chronic hepatitis B patients with HBsAg+/HBsAb+ were used as experimental group,and specimens collected from 35 cases of newly diagnosed chronic hepatitis B patients with HBsAg+/HBsAb-were used as control group.The S gene region of HBV DNA was amplified and sequenced.The amino acid sequences were classified to different genotypes and the mutations were analyzed based on the sequencing results. Results The experimental group contained 8 patients of the B genotype and 35 patients of the C genotype,whereas the control group contained 9 patients of the B genotype and 26 patients of the C genotype.In experimental group,the average age of the C genotype patients [(50.2±16.3) years] was obviously older than that of the B genotype patients [(34.4±13.4) years](P=0.015).The mutation rates for the B and C genotypes in the experimental group were 0.77% and 1.64%,respectively(P=0.005).Whereas the mutation rates were 0.59% and 0.49% for the B and C genotypes in control group(P=0.597).In addition,among the C genotype patients,the experimental group had a marked increase in S gene amino acid mutations,and a higher amino acid mutation rate was observed in the major hydrophobic region(MHR,aa100～169),especially in the first loop(aa124～137) of α determinant(aa124～147).However,only few patients with the B genotype had mutations in α determinant.Conclusion The C genotype patients with HBsAg+/HBsAb+ are likely to have more mutations than the B genotype patients,especially the mutations in α determinant.The mutations in this area may lead to changes of antigenicity and simultaneous HBsAg+/HBsAb+ finally.