[Effect of immunotherapy of recombinant chimeric epitopes of major allergen group 1 from Dermatophagoides farina on asthma of mice].

OBJECTIVE: To investigate the effect of immunotherapy of recombinant chimeric epitopes of major allergen group 1 from Dermatophagoides farina on asthma of mice. METHODS: Forty mice were randomly divided into 4 groups: a negative control group, an asthma group, an immunotherapy group of Der f 1, and an immunotherapy group of Der f lA. On the 1st, 7th and 14th day, the mice in the asthma group, immunotherapy group of Der f 1, and immunotherapy group of Der f 1A were injected intraperitoneally with the extract of D. farina 3 times to sensitize; and on the 21st day, the atomized inhalation was carried out for 7 days. In the control group, phosphate buffer solution (PBS) was applied for sensitization and inhalation. In the immunotherapy groups, Der f 1 and Der f 1A were applied to carry out the specific immunotherapy respectively for 30 min before the inhalation. Then, the leukocytes in the bronchoalveolar lavage fluid (BALF) were numbered and the pathological sections of lung tissues were observed; IL-5 and IFN-γ in BALF and spleen cell culture supernatants (SCCS) as well as the specific IgE, IgG2a in the sera were detected. RESULTS: Compared with the asthma group, the lung inflammation of mice in the immunotherapy groups was lightened, and the total numbers of leukocytes in BALF were significantly reduced; IL-5 was significantly reduced and IFN-γ was significantly increased in BALF and SCCS of mice in the immunotherapy groups; and the specific IgE was significantly reduced and IgG2a was significantly increased in the sera of mice in the immunotherapy groups (all P< 0.01). CONCLUSION: The recombinant chimeric epitopes of major allergen group 1 from D. farina could effectively relieve the symptom of asthma in mice, so as to provide the evidence for specific immunotherapy.

[Construction and expression of a chimeric gene with T-/B-cell epitopes of major allergen group 1 from Dermatophagoides farina].

OBJECTIVE: To construct and express a chimeric gene with T-/B-cell epitopes of the major allergen group 1 from Dermatophagoidesfarina (Der f 1). METHODS: Two chimeric genes, Der f 1A and Der f 1B, were synthesized as B1-T1-B2-T2- B3-T3-B4-T4-B5-T5-B6 and B1-B2-B3-B4-B5-B6-T1-T2-T3-T4-T5 pattens. Two recombinant vectors, pET-28a(+)-Der f 1A and pET-28a(+)-Der f 1B, were constructed for prokaryotic expression. These chimeric genes were induced by 1 mmol/L IPTG (final concentration), digested with restriction enzymes and sequenced. The chimeric proteins were analyzed by SDS-PAGE and Western blotting. RESULTS: After digestion by restriction enzymes and sequencing, the recombinant vectors were constructed successfully. The specific bands for chimeric proteins were visible by SDS-PAGE and Western blotting, suggesting that these proteins were purified successfully. Further analyses were performed for IgE-binding properties of Der f 1A and Der f 1B to sera from patients sensitized to house dust mite. Compared with the parental allergens Der f 1, Der f 1A and Der f 1B had reduced IgE -binding capacity (both P < 0.05), whereas the difference between Der f 1A and Der f 1B was not statistically significant (P > 0.05). CONCLUSION: Two chimeric proteins are expressed successfully, which contain T-/B-cell epitopes of Der f 1 and provide a basis for specific immunotherapy.

[Experimental study on polysaccharide of Cipangopaludina chinensis against HBV in vitro].

OBJECTIVE: To evaluate the biological activity of polysaccharide of Cipangopaludina chinensis (PCC) against HBV in vitro. METHOD: HepG2 2. 2. 15 cells were taken as the in vitro experimental model. The cell toxicity was observed by MTT. PCC of different safe concentrations and positive control medicine 3TC were added into the cells. Cell control without medicine was set at the same time. Cultural supernatants were collected at 9 d. HBsAg and HBeAg in cultural supernatants were tested by ELISA. The content of HBV-DNA was detected by TaqMan probe fluorescence quantitative PCR. RESULT: TC0 and TC50 of PCC in HepG2 2. 2. 15 cell culture were 1 g . L-1 and >10 g . L-1, respectively, suggesting low toxicity in cells. IC50 of PCC in HepG2 2. 2. 15 cells HBsAg and HBeAg were 0. 501, 0. 401 g. L-1, with SI being >19.96 and >24. 94, respectively. PCC could effectively inhibit the secretion of HBsAg and HBeAg, and have a better effect on HBeAg than on HBsAg. PCC had a significant inhibitory effect on HBV-DNA in HepG2 2. 2. 15 cells at concentrations of 0. 1, 1 g . L-1 P <0.05). CONCLUSION: PCC has the effect against HBV activity in vitro to some extent, with low toxicity, thereby having a good prospect for application.

[Human enterovirus 71 (EV71) resisted type I interferon induced antiviral effect].

OBJECTIVE: To investigate human enterovirus 71 (EV71) resistance to type I interferon induced antiviral effect. METHODS: After type I interferons (alpha, beta) were incubated with HeLa cells, recombinant type I herpes simple virus (HSV-1) with green fluorescent protein (GFP) was inoculated onto the HeLa cells. HSV-1 proliferation was observed by GFP expression and PCR. After EV71 was inoculated onto HeLa cells incubated with the same quantity of interferon, proliferation of EV71 were detected by RT-PCR of 2A gene. RESULTS: Recombinant HSV-1 GFP expression and viral DNA replication obviously decreased in HeLa cells incubated with type I interferon (alpha, beta). However, EV71 effectively proliferated in the interferon irritated HeLa cell by RT-PCR. CONCLUSION: HeLa cell irritated by type I interferon (alpha, beta) produced antiviral substance that inhibits HSV-1 proliferation. EV71 resisted the antiviral substance induced by type I interferon and could significantly replicate in the HeLa cells.

[Development of a GeXP based multiplex RT-PCR assay for simultaneous detection of eight arboviruses related to encephalitis].

Multiplex reverse transcription-polymerase chain reaction (mRT-PCR) is currently available in virus detection and defined as the simultaneous amplification of two or more DNA/RNA targets in a single reaction vessel. In this study, we attempted to modify the conventional mRT-PCR technique on a basis of GenomeLab Genetic Analysis System (GeXP). Initially, we optimized the analytical validation of the GeXP analyzer and its design of workflow and simultaneously detected eight arboviruses that related to epidemic encephalitis by verifying the specificity of mRT-PCR with Japanese encephalitis virus(JEV) cell cultures and positive strains identified previously and determining the sensitivity with in vitro-transcribed RNA of serial dilutions. The GeXP system after optimization could amplify the specific fragments related to the viruses and exposed specifically a total of 13 target genes out of eight types of arboviruses at the level of 10(2) copies/microL, and the findings suggest that the novel protocol we developed can be high-throughput and highly specific and sensitive as well as quickness in screening of the encephalitis viruses, and is promising in detection of encephalitis-associated viruses for molecular epidemiological studies.

[Molecular interaction between PrP protein and the signal protein 14-3-3 beta].

The molecular interaction between PrP and 14-3-3 beta and the possible interactional domain between two proteins were studied by co-immunoprecipitation, pull down and FRET assays. The results showed that PrP protein could interact with 14-3-3 beta in vitro and in vivo. The domain which responded for the interaction was located at C-terminal of PrP (amino acid residues 106 to 126). This study of the interaction between PrP and 14-3-3 protein further provided the insight into the potential role of 14-3-3 in the biological function of PrP and the pathogenesis of prion disease.

[Study on the phenomenon of splashes and sprays from virology].

OBJECTIVE: To investigate the phenomenon of accidental splashes and sprays from manipulation of recombinant virus material and to measure the approximate spilled distance when recombinant virus material inadvertently dropped in the biosafety laboratory. METHODS: first, two groups owning different experience simulated the course of accidental spills and splashes by recombinant adenovirus (rADV) which expressed green fluorescence protein (GFP), the GFP signal were observed in 96 well cell plate after spills appeared; Second, the routine two heights (75 cm and 110 cm) and capacity (1 ml, 1.5 ml, 4 ml and 8 ml) of virus were chose to simulate the experiment of unexpected dropping. RESULTS: First, the positive quantity of the first group owning 5 years' experience is much less than the second group owning 2 years' work experience, the former was 7 positive wells, the latter was 81 positive when they used the pipette to operation. Second, when the unclosed test tubes (1 ml, 1.5 ml, 4 ml and 8 ml recombinant virus) inadvertently dropped, the largest spill distance was 0.92 m, 1.57 m, 2.63 m and2.68 m respectively. CONCLUSION: The better experience is important to make sure safety when we make infectious material; the contaminated distance increased with the amount of recombinant virus material.

[Study of Scutellaria baicalensis and Baicalin against antimicrobial susceptibility of Helicobacter pylori strains in vitro].

OBJECTIVE: To compare the antimicrobial activities of Scutellaria baicalensis and Baicalin against Helicobacter pylori (H. pylori) in vitro. METHODS: The crude alcohol extraction of Scutellaria baicalensis was obtained by successive extractions with ethanol. Baicalin was extracted by using organic extraction methods and the purity was analyzed by high-performance liquid chromatography (HPLC). The crude alcohol extraction of Scutellaria baicalensis and Baicalin were used in broth dilution assays to test for antibacterial properties. RESULTS: The content of Baicalin was 5.01 g from 100 g Scutellaria baicalensis. The purity of Baicalin was 96.8%. The crude alcohol extraction of Scutellaria baicalensis and Baicalin were tested for their ability to inhibit H. pylori in vitro by using broth dilution assays. The MIC50 and MIC90 of Baicalin against ten strains of H. pylori were 1.04 and 1.30 mg/ml respectively. The MIC50 and MIC90 of the crude product against ten strains of H. pylori were 2.60 and 3.26 mg/ml respectively. CONCLUSION: The Baicalin and Scutellaria baicalensis are bactericidal against H. pylori. The antimicrobial activity of Baicalin is greater than that of Scutellaria baicalensis.

[The construction and identification of the PRNP vectors with ubiquitin or the lysosome-targeting signal].

OBJECTIVE: To evaluate PrP expression characteristic of PRNP nucleic acid vaccine vector with ubiquitin or the lysosome-targeting signal. METHODS: The gene of ubiquitin and lysosome-targeting signal were ligated to PRNP and pcDNA3.1 vector that is, pcDNA3.1-UPrP and pcDNA3.1-PrPL were constructed. The expression characteristics of PrP with two signals were evaluated by Western Blot and the localization was observed by indirect immune fluorescence. RESULTS: The protein expressed by pcDNA3.1-UPrP and pcDNA3.1-PrPL with ubiquitin and lysosome-targeting signal can be recognized by prion-specific antibody. The protein has three glycosylation molecules form as native PrP.PrP with ubiquitin was degraded gradually with time extension,whereas quantity of PrP with lysosome signal reduced in 48 h after transfection. The protein with two location signals can direct fusion proteins to cytoplasm. CONCLUSION: The PRNP vectors with ubiquitin or the lysosome-targeting signal were constructed and expressed in eukaryocyte successfully. There will be one of good foundation on PRNP nucleic acid vaccine.

[Investigating the risk of two model virus in the laboratory].

OBJECTIVE: To study the survival time of recombination rival in environment and inactivation ability of different disinfectant and ultraviolet radiation against virus. METHODS: NC membranes absorbed the recombinant adenovirus (rADV) or herpes simplex virus (rHSV) with green fluorescence protein (GFP) were laid, or immersed in various concentration of different disinfectants such as ethanol, sodium hypochlorite, lysol and geramine and then taked out them every 15 min, or exposed under ultraviolet radiation, then the NC membranes were adsorbed 1 h in cell, 37 degrees C 5% CO2 48 h. The results were observed under the fluorescence microscope. RESULTS: (1) the average survival time of rHSV under environment is less than 60 min, rADV is almost up to 2 h. (2) The infection ability of rHSV and rADV was inactived 15 min by both ethanol (100%, 70% and 50%) and sodium hypochlorite (5%, 2.5% and 1.25%). (3) Two virus can be killed by 0.1% bromogeramine. (4) Both 5% and 2.5% lysol, but rADV can not lost the infection on Vero Cell until 75 min by 1.25% Lysol. (5) The rHSV was inactivated under ultraviolet radiation, but rADV was not. CONCLUSION: The survival time of is different from both envelope rival and the no-envelope viral under nature environment and the inactivate ability of disinfectant also is different between two model virus; Disinfectant should be choose according to virus type.

[Survey on natural nidus of Trichobilharzia in Huainan area].

The life cycle of Trichobilharzia sp. can be completed in Radix auricularia and domestic or wild ducks, and people can contract cercarial dermatitis through water contact. Natural nidus of Trichobilharzia exists in Huainan area.

Effect of liniment levamisole on cellular immune functions of patients with chronic hepatitis B.

AIM: To explore the effects of liniment levamisole on cellular immune functions of patients with chronic hepatitis B. METHODS: The levels of T lymphocyte subsets and mIL-2R in peripheral blood mononuclear cells (PBMCs) were measured by biotin-streptavidin (BSA) technique in patients with chronic hepatitis B before and after the treatment with liniment levamisole. RESULTS: After one course of treatment with liniment levamisole, the levels of CD3(+), CD4(+), and the ratio of CD4(+)/CD8(+) increased as compared to those before the treatment but the level of CD8(+) decreased. The total expression level of mIL-2R in PBMCs increased before and after the treatment with liniment levamisole. CONCLUSION: Liniment levamisole may reinforce cellular immune functions of patients with chronic hepatitis B.

Cloning and sequencing of cagA gene fragment of Helicobacter pylori with coccoid form.

AIM: To clone and sequence the cagA gene fragment of Helicobacter pylori (H pylori) with coccoid form. METHODS: H pylori strain NCTC11637 were transformed to coccoid form by exposure to antibiotics in subinhibitory concentrations. The coccoid H pylori was collected. cagA gene of the coccoid H pylori strain was amplified by PCR. After purified, the target fragment was cloned into plasmid pMD-18T. The recombinant plasmid pMD-18T-cagA was transformed into E.coli JM109. Positive clones were screened and identified by PCR and digestion with restriction endonucleases. The sequence of inserted fragment was then analysed. RESULTS: cagA gene of 3,444 bp was obtained from the coccoid H pylori genome DNA. The recombinant plasmid pMD-18T-cagA was constructed, then it was digested by BamH I+Sac I, and the product of digestion was identical with the predicted one. Sequence analysis showed that the homology of coccoid and the reported original sequence H pylori was 99.7%. CONCLUSION: The recombinant plasmid containing cagA gene from coccoid H pylori has been constructed successfully. The coccoid H pylori contain completed cagA gene, which may be related to pathogenicity of them.

Helicobacter pylori L-form and patients with chronic gastritis.

AIM: To study the relationship between infection with Helicobacter pylori L-forms and chronic gastritis and its association with possible changes of cellular immune function. METHODS: Gastric mucosal biopsies were taken from 428 patients with chronic gastritis to detect H pylori L-form by Gram staining and immunohistochemistry staining. Peripheral venous blood samples of patients were taken to detect the percentage of CD3+, CD4+ and CD8+ by the biotin-streptavidin (BSA) assay and the levels of IL-2, IL-6 and IL-8 by ELISA. RESULTS: The rate of infection with H pylori L-forms was 48.83% (209/428). The rate was 50.47% (216/428) and 52.80% (226/428), respectively, as detected by immunohistochemistry staining and Gram staining (P>0.05). The rate of H pylori L-forms in males and females was 57.8% (136/235) and 37.28% (73/193), respectively, (chi2=17.05, P<0.01). Furthermore, the rate increased with age, with the rate being significantly greater in patients > or =40 years old than in those <40 years old (P<0.01). The percentage of CD3+, CD4+, CD8+, the ratio of CD4+/CD8+, and the levels of IL-2, IL-6, IL-8 in H pylori-positive patients were 47.58+/-4.44%, 25.51+/-4.74%, 22.77+/-7.46%, 1.44+/-0.51%, 1.56+/-0.47 mg/L, 103.62+/-5.85 ng/L, and 109.79+/-7.18 ng/L, respectively. Compared with H pylori-negative patients, the percentage of CD3+, CD4+ and the ratio of CD4+/CD8+ and the IL-2 level decreased, but the levels of IL-6, IL-8 increased (P<0.001-P<0.01). Moreover, the percentage of CD3+, CD4+, CD8+, the ratio of CD4+/CD8+, and the levels of IL-2, IL-6 and IL-8 in patients infected with both H pylori L-forms and vegetative forms were 46.67+/-5.21%, 30.75+/-4.89%, 22.15+/-6.45%, 1.32+/-0.47%, 1.16+/-0.38 mg/L, 116.45+/-5.44 ng/L, and 118.64+/-6.24 ng/L, respectively. Compared with patients infected with only vegetative forms, the percentage of CD4+, the ratio of CD4+/CD8+and IL-2 level decreased, but the levels of IL-6 and IL-8 increased (P<0.001 or P<0.01). CONCLUSION: L-form variation often occurs in patients with chronic gastritis and is commonly found in male patients and associates with ages. The L-form variation may be an important factor causing disorder of cellular immune function in the patients with H pylori-induced chronic gastritis.

Clinical and epidemiological features of patients with clonorchiasis.

AIM: To study the clinical and epidemiological features of patients with clonorchiasis so as to provide scientific evidences for the diagnosis and prevention of clonorchiasis. METHODS: Stools from 282 subjects suspected of having clonorchiasis were examined for helminth eggs with modified Kato's thick smear and sedimentation methods, and their sera were tested for HAV-DNA, HBV-DNA, HCV-RNA, HDV-RNA and HEV-RNA with polymerase chain reaction (PCR). Clinical symptoms of patients with clonorchiasis only were analyzed, and their blood samples were tested for circulating antigen (CAg) with Dot-ELISA, eosinophilic granulocyte count, and alanine aminotransferase (ALT). Meanwhile, they were asked to provide data of occupation, eating habit, hygienic habit and knowledge of clonorchiasis. In addition, the ecosystem of the environment in epidemic areas was surveyed. RESULTS: Among the 282 patients, 61 (21.43%) were infected with clonorchis sinensis only, 97 (34.64%) were co-infected with clonorchis sinensis and other pathogens, 92 (32.86%) were infected with hepatitis virus only and 31 (11.07%) neither with clonorchis sinensis nor hepatitis virus. Among the 61 patients with clonorchiasis only, there were 14 (22.95%) subjects with discomfort over hepatic region or epigasfrium, 12 (19.67%) with general malaise or discomfort and inertia in total body, 6 (9.84%) with anorexia, indigestion and nausea, 4 (6.56%) with fever, dizziness and headache (6.56%), and 25 (40.98%) without any symptoms; sixty one (100%) with CAg (+), 98.33% (59/60) with eosinophilic granulocytes increased and 65.00% (39/60) with ALT increased. B-mode ultrasonography revealed 61 cases with dilated and thickened walls of intrahepatic bile duct, and blurred patchy echo acoustic image in liver. Twenty-six cases had stones in the bile duct, 39 cases had slightly enlarged liver with diffuse coarse spots in liver parenchyma. Twenty cases had enlarged gallbladder with thickened coarse wall and image of floating plagues, 9 cases had slightly enlarged spleen. By analysis of epidemiological data, we found that the ecologic environment was favorable for the epidemiology of clonorchiasis. Most patients with clonorchiasis were lack of knowledge about the disease. Their living environment, hygienic habits, eating habits and their occupations were the related factors that caused the prevalence of the disease. CONCLUSION: The clinical symptoms of clonorchiasis are non-specific, and the main evidences for diagnosis of clonorchiasis should be provided by etiologic examination, B-mode ultrasonography and clinical history. The infection of clonorchis sinensis is related to occupations, bad eating habits and lack of knowledge about prevention of the disease.

Cloning and expression of vacA gene fragment of Helicobacter pylori with coccoid form.

BACKGROUND: Helicobacter pylori (H. pylori) can transform, in vivo as well as in vitro, from dividing spiral-shaped forms into nonculturable coccoid forms, whose importance in disease transmission and antibiotic treatment failures is unclear. The aim of the present study was to clone and express the vacA gene of coccoid H. pylori and to infer its possible pathogenesis. METHODS: Firstly, coccoid form was obtained from strain NCTC 11637 by exposure to antibiotics in subinhibitory concentrations and collected. Secondly, vacA gene of the coccoid H. pylori was amplified by PCR. After being purified, the target fragment was cloned into plasmid pMD-18T, and the recombinant plasmid pMD-18T-vacA was transformed into E. coli JM109. The sequence of inserted fragment was analyzed. Thirdly, vacA gene from recombinant plasmid pMD-18T-vacA was digested with restriction enzyme and was inserted into expression vector pET32a (+). The positive recombinants were transferred into E. coli BL21 and identified by restriction enzyme digestion and PCR. Finally, the genetically engineered bacteria including pET32a (+)-vacA plasmids were induced by IPTG, the expression was analyzed by SDS-PAGE and gel densitometric scanning. RESULTS: The results revealed that vacA gene of 3888bp was obtained from the coccoid H. pylori genome DNA, recombinant plasmid pMD-18T-vacA constructed were successfully digested by BamH I +Sac I, and the product of digestion was identical with the predicted 1. Sequence analysis also showed that the homology of coccoid and the reported original sequence was 99.8%. Plasmid pET32a (+)-vacA could express a specific 156kDa protein in E. coli BL21, and the protein accounted for 15.5% of the total protein of recombinant bacterial. CONCLUSIONS: The present data indicate that coccoid H. pylori contains complete vacA gene, and could synthesize its protein, which may be related to the disease relapse and transmission when coccoid H. pylori recovers virulence under suitable conditions.

Detection of serum anti-Helicobacter pylori immunoglobulin G in patients with different digestive malignant tumors.

AIM: To investigate the seroprevalence of Helicobacter pylori infection in patients with different digestive malignant tumors. METHODS: Enzyme linked immunosorbent assay (ELISA) was used to detect serum anti-Helicobacter pylori IgG antibody in 374 patients with different digestive malignant tumors and 310 healthy subjects (normal control group). RESULTS: The seroprevalence of Helicobacter pylori infection was 61.50% (230/374) and 46.77% (145/310), respectively, in patients with digestive tumors and normal controls (P<0.05). The seroprevalence was 52.38% (33/63), 86.60% (84/97), 83.14% (84/101), 45.24 (19/42), 51.13% (18/35) and 44.44% (16/36), respectively in patients with carcinomas of esophagus, stomach, duodenum, rectum, colon and liver (P<0.01). In patients with intestinal and diffuse type gastric cancers, the seroprevalence was 93.75% (60/64) and 72.73% (24/33), respectively (P<0.05). In patients with gastric antral and cardiac cancers, the seroprevalence was 96.43% (54/56) and 73.17% (30/41), respectively (P<0.05). In patients with ulcerous and proliferous type duodenal cancers, the seroprevalence of H pylori infection was 91.04% (61/67) and 52.27% (23/44), respectively (P<0.05). In patients with duodenal bulb and descending cancers, the seroprevalence was 94.20% (65/69) and 45.20% (19/42), respectively (P<0.05). CONCLUSION: H pylori infection is associated with occurrence and development of gastric and duodenal carcinomas. Furthermore, it is also associated with histological type and locations of gastric and duodenal carcinomas.

Detection of T lymphocyte subsets and mIL-2R on surface of PBMC in patients with hepatitis B.

AIM: To study the levels of T lymphocyte subsets and membrane interleukin-2 receptor (mIL-2R) on surface of peripheral blood mononuclear cells (PBMCs) of patients with hepatitis B and its role in the pathogenesis of hepatitis B. METHODS: The levels of T lymphocyte subsets and mIL-2R in PBMC before and after being stimulated with PHA were detected by biotin-streptavidin (BSA) technique in 196 cases of hepatitis B. RESULTS: In patients with hepatitis B, the levels of CD(3)(+), CD(4)(+) cells, and the ratio of CD(4)(+) cells/CD(8)(+) cells were lower, but the level of CD(8)(+) cells was higher than those in normal controls (42.20+/-6.01 vs 65.96+/-6.54, 38.17+/-5.93 vs 41.73+/-6.40, 0.91+/-0.28 vs 1.44+/-0.31, 39.86+/-6.36 vs 30.02+/-4.54, P<0.01). The total expression level of mIL-2R in PBMC before and after being stimulated with PHA was also lower than those in normal controls (3.47+/-1.55 vs 4.52+/-1.49, 34.03+/-2.94 vs 37.95+/-3.00, P<0.01). In all the patients with hepatitis B, the levels of T lymphocyte subsets and mIL-2R in PBMC with HBV-DNA (+) were lower than those with HBV-DNA (-), which were significantly different (39.57+/-7.11 vs 44.36+/-5.43, 34.36+/-7.16 vs 40.75+/-5.87, 37.82+/-6.54 vs 41.72+/-6.21, 0.88+/-0.33 vs 0.99+/-0.27, 2.82+/-1.62 vs 3.85+/-1.47, 31.56+/-3.00 vs 35.84+/-2.83, P<0.01). In addition, the levels of CD(3)(+), CD(4)(+), CD(8)(+) cells, the ratio of CD(4)(+) cells /CD(8)(+) cells and mIL-2R among different courses of hepatitis B were all significantly different (F=3 723.18, P<0.01. F=130.43, P<0.01. F=54.01, P<0.01. F=2.99, P<0.05. F=7.16, P<0.01). CONCLUSION: Both cellular and humoral immune functions are obviously in disorder in patients with hepatitis B, which might be closely associated with the chronicity in patients.