Objective To investigarte the immune status in the patients with dengue fever .Methods The flow cytometry was used to detect the T lymphocytes in the patients with dengue fever for analyzing their immune status .Results Compared with the reference range in the healthy individuals ,it was found that the T lymphocyte proportion in the patients with dengue fever was sig‐nificantly reduced compared with the healthy individuals .The percentages of CD3+CD4+ and CD3+CD8+ lymphocytes were (36 .54 ± 9 .78)% and (17 .7 ± 10 .01)% respectively ,which had statistical difference compared with the control group(P<0 .05) ,CD3+CD4+ lymphocyte count was (49 .98 ± 240 .2)cells/μL ,the difference was statistically significant (P<0 .05) ,CD3+ CD8+ lympho‐cytes count was (380 .9 ± 364 .6)cells/μL ,the difference was statistically significant(P<0 .05) .Conclusion The immune status in the patients with dengue fever is abnormal ,T lymphocyte percentage is significantly reduced compared with the healthy individuals .

detection of HIV-1 quasispecies in HIV-1 infected populations with low level viral load.

Objective To investigate the characteristics of V3 loop amino acid sequences of human immunodeficiency virus type 1 (HIV-1) quasi-species in long-term non-progressors (LTNP)infected with HIV. Methods End-point limiting dilution polymerase chain reaction (EPLD PCR) was used to amplify the env gene c2-v3-c3 region of single HIV-1 provirus from five LTNPs at sequential time points. The PCR products were then sequenced and the amino acid sequences of V3 loop were analyzed by sequence confirm analysis technology. Results The results showed that there were one to ten kinds of polymorphisms in the V3 region of HIV-1 quasi-species which were found from the serial samples of the five LTNP. However, the sequences of the predominant strains were either completely consistent or at most changed at one or two residues in the serial samples of individual patient. The tetramer compositions of the tip of V3 loop were consistent in each patient. It was GPGR in four patients and GPGK in one patient. It was speculated the co-receptor of HIV-1 was CC chemokine receptor (CCR)-5 based on the amino acids at the residue 11 and residue 25 of V3 loop and the net charge of V3 loop. Conclusions There are various polymorphisms at the HIV V3 loop in LTNP. However, the tetramer composition of the tip part of V3 loop is stable. The LTNP are very likely infected with non-syncytium inducing (NSI) strain.

Objective To identify the pathogens that cause hand, foot and mouth disease (HFMD) in adults and analyze the nucleotide sequences characteristics of enterovirus 71 (EV71). Methods The reverse transcription-polymerase chain reaction (RT-PCR) method was used to detect the enterovirus from the samples of four adult HFMD patients. The 227 bp amplified segments of EVT1 were then sequenced and compared with the sequences of previously isolated EVT1 strains available from GenBank by homogeneity and phylogenetic tree analyses. Results All the results of RT-PCR with enterovirus universal primers and EVT1 specific primers were positive. The EV71 sequences analysis showed that the four new sequences (named as GZ19610, GZ99310, GZ99355 and GZ46477) shared 96.0% to 99.1% nucleotide identify themselves and shared 96.9% to 100.0% homology with the strain Fuyang/17.08/3 isolated in 2008 from Fuyang, Anhui Province. Phylogenetic tree analysis showed that the genotype of the four new sequences was all subtype C4, they were the same sub-genotype as those strains isolated from Chinese mainland and Chinese Taiwan in 2004, and the genetic distance between them was most closely. Conclusions EV71 can cause adult HFMD. Compared with the nucleotide sequences of EV71 strains that isolated now and formerly in China, there is no large variation of the EV71 sequences isolated from four adult HFMD patients in Guangzhou this time. The adult HFMD patients should be isolated for treatment to avoid them transmitting the virus and causing disease spreading.

Objective:To study the characteristics of chromogranin(CgA),synaptophysin(SYN) and neuron-specific enolase(NSE) in the cortex of adrenal gland.Methods:Immunohistochemical technique was used to detect the expressions of CgA,SYN and NSE in 56 routinely processed tissue specimens from human adrenal cortical neoplasm and hyperplasia.Results: CgA immunoreactivity was regularly detected in the cortex of adrenal neoplasm,but not in the hyperplasia and normal adrenal gland.The immunoreactive materials appeared in the cytoplasm and in the form of vacuole or grains.Adrenal cortical neoplasm and hyperplasia showed NSE positive cells in the cortex,but not in the normal adrenal gland.SYN positive materials were shown in the cortex of hyperplasia and adrenocorticoadenoma,but not in the adrenocorticoadenocarcinoma and normal adrenal gland.Conclusion: The distributive characteristics of CgA,SYN and NSE in the adrenal cortex help the differential diagnosis of adrenal hyperplasia.The adrenal cortex is closely connected with medulla.Besides the classical hypothalamus-pituitary-adrenal axis(HPAA) and rennin-angiotensin-aldosterone system(RAAS),the adrenal cortex and medulla are mutually regulated on the basis of anatomy and endocrinology.

Objective To study the characteristic of chromogranin A(CgA),synaptophysin(SYN) and neuron-specific enolase(NSE) in cortex and medulla of adrenal.Methods Immunohistochemical method was used to detect the expression of CgA,SYN and NSE in routinely processed tissue specimens from human adrenal neoplasm and hyperplasia.Results The expression of CgA was detected in cortex of adrenocortical adenoma,adrenal neoplasm and chromophile tumor,but not in that of hyperplasia and normal adrenal.The expression of NSE was detected in cortex of adrenal cortical hyperplasia,cortical adenomas and pheochromocyte carcinoma,but not in that of pheochromocytoma and normal adrenal.The expression of SYN was detected in cortex of corticohyperplassia,cortical adenomas,medull neoplasm,chromophile tumor and pheochromocyte carcinoma,but not in that of cortical adenocarcinoma and normal adrenal.The expressions of CgA,NSE and SYN were detected in the medulla of all cases.Conclusion The characteristic of CgA,SYN and NSE in cortex and medulla of adrenal are useful in the differential diagnosis of tumor and hyperplasia in the adrenal.The relationship between the adrenal cortex and medulla is close.Besides the classical hypothalamus-pituitary-adrenal axis(HPAA) and rennin-angiotensin-aldosterone system(RAAS),the adrenal cortex and medulla have the basis of anatomy and endocrinology.

Objective To investigate the relationship between HBV mutations in the precore (PC)/core promoter region and the liver histological changes in HBeAg negative CHB patients. Method A total of 71 HBeAg negative CHB patients with liver biopsy from April 2012 to Dec 2013 were enrolled. DNA was extracted from blood serum, then the HBV S gene and PC/core promoter region were amplified by semi-nested PCR and sequenced. The relationship between significant liver histological changes and viral factors were analyzed by Logistic regression analysis. Results The incidence of significant necroinflammation (15.8% vs. 27.3%, χ2 =1.398, P = 0.237) and significant fibrosis (71.1% vs. 84.4%, χ2= 1.926, P = 0.165) were found to be similar between patients infected with HBV genotype B and genotype C . By Logistic regression analysis including risk factors of age, sex, HBV genotype and mutations (T1753V,A1762T/G1764A,A1846T and G1896A), the A1762T/G1764A mutation in HBV associated with significant necroinflammation (OR = 4.296, P = 0.037), while factors of age, sex, genotype and other mutation were not associated with significant liver histological changes. (all P > 0.05). Conclusion Mutation in PC/core promoter region of HBV may act as a marker to evaluate the liver histological changes.

Objective To investigate a non-invative , sensitive and specific method to diagnose bladder cancer, and evaluate the value of the combination of FISH with NMP22 in the diagnosis of bladder cancer. Methods Urine from 68 patients suspected suffering from bladder cancer were used by FISH, NMP22 expression and cytologi-cal examination, respectively. The results of above detections were compared to the subsequent histopathology as-say to analyze the specicficity, sensitivity of diagnosis for bladder cancer. Results The sensitivity of FISH, NMP22 expression and the cytological examination was 81.4%,86.0% and 39.5% respectively, and the specifici-ty of FISH, NMP22 expression and the cytological examination was 84.0%, 72.0% and 96.0%, respectively. The sensitivity and specificity of the combination of FISH with NMP22 was 79.1% and 92.0%. Conclusions The combined diagnosis of FISH and NMP22 for bladder cancer showed greater sensitivity than that of the conventional cytology, with similar specificity as the conventional cytology. The combination of FISH with NMP22 test shows more value for the diagnosis of bladder cancer than any single assay.

Background and purpose: The clinical and pathological characteristics and treatment strategy for the patients with testicular diffuse large B cell lymphoma still need to be further studied. This study was done to evaluate the diagnosis and strategy for the disease by retrospective analysis of 21 patients with testicular diffuse large B cell lymphoma. Methods: 21 patients with pathologically proved as diffuse large B cell lymphoma after surgery in the Sun Yat-sen University Cancer Center and The Guangzhou General Hospital of PLA from September 2002 to April 2009 were accrued. There were 3 in stage Ⅰ_E , 4 in stage Ⅱ_E , 5 in stage Ⅲ_E and 9 in stage Ⅳ according to Ann Arbor stage standard. All patients received adjuvant chemotherapy with CHOP regimen initially after operation. Radiotherapy or other salvage chemotherapy regimen was given after failure of the initial treatment. The follow-up information was collected for to all of the patients. Results: Follow-up time ranged from 10 to 83 months. After completion of first-line chemotherapy(CHOP), overall response rate was 72.2%(13/18)with complete remission (CR) rate of 33.3%(6/18)and partial response rate(PR) of 38.9%(7/18). 3 patients in stage Ⅰ_E survived free from disease until now. Tumor relapsed and refractory eases were observed in 10 patients, and they all showed little response to chemotherapy with a response rate of 30.0%(3/10). Among the 21 patients, 11(52.3%) cases died of the disease with a median survival time of 28 months. Of these 21 patients, 7 patients were CD10(+), 5 patients overexpressed bel-6, 12 patients were bcl-2(+), and 15 patients overexpressed MUM1.Overexpressed bcl-2 and MUM1 indicated bad prognosis. Conclusion: Most of the testieular diffuse large B cell lymphoma patients were non-GCB. And they were sensitive to CHOP chemotherapy. The results suggested that all patients should receive chemotherapy after surgery. It was curable in the early stage. However,the options of treatment method for the patients with relapsed and refractory cases are limited and need to be further explored.

Objective To identify the sequence of hepatitis B virus S gene"a"determinant in patients with positive HBsAg and anti-HBs.Methods Nested-PCR Was used to amplify the HBV S gene in 4 patients with positive HBsAg and anti-HBs,and the PCR products were sequenced directly or after cloning.The sequences of"a" determinants were then analyzed by sequence alignment.Results Direct sequencing of PCR products showed that there was one amino acid (aa)residue in"a"determinant less conserved region emerging polymorphism in all 4 patients.Clone sequencing showed that aa residue at 126 of "a"determinant in patient 1 miSht be Thr,Ile and Set,at 134 might be Phe and Set;the aa at 126 in patient 2 misht be Ala and Thr.and in patient 3 might be Ile and Asn;aa polymorphism was not found in patient 4.Conclusion The polymorphism of"a"determinant in HBV S gene might be associated with positivity of both HBsAg and anti-HBs in hepatitis B patients.