Objective To investigate the significance of human cyomeg Movirus(HCMV)pp65 IgG antibody avidity index(AI)for the clinical diagnosis of HCMV primary infection through the experimental model of HCMV primaly infection in BALB/c mice.Methods 6～8 weeks,female,specific-pathogen-free BALB/c mice were divided into 5 groups.6 mice in each group. And he mice were injected with 2×106 PFU/m1,2×105 PFU/mi,2×104 PFU/ml,2×103 PFU/ml and 2×102 PFU/ml of HCMV intraperitoneally respectively. Another 6 mice were injected intraperitoneally with the maximum dose of HCMV kept at 56℃ for 30 min as inactivated virus group.And HF negative control group was established at same time. All the mice ere sacrificed to obtain brain and lung tissues for the following experiments after 1 montll.(1)Tissue samples obtained from mice were inoculated in human embryo fibroblasts(HF)monolayers after routine treatment for virus isolation.HCMV specific eytopathie effect(CPE) was observed bv inverted phase-contrast microscopy.HCMV UL83 DNA in the ultures as tested by PCR and pp65 antigen was detected by indirect immunofluorescence.(2)Extracted mRNA from tissue samples and HCMV pp67 mRNA were analyzed by reverse transcriptase PCR(RT-PcR).(3)Immunoglobulin M(IgM)antibody and immunoglobulin G(1gG)antibody avidity Was investigated for their usefulness in distinguishing primary genital HCMV infections rom nonprimary infections with ELISA kit using truncated pp65 protein.ResultsHCMV can be isolated in the tissues from the mice injected with 2×106 PFU/ml and 2×105 PFU/m1.RT- PCR and ELISA showed positive results in the same groups.The infective rates were 100%.The analysis of the low doses groups,inactivated group and HF negative ontrol group all showed negative results.Conclusions BALB/c mice can be infected with HCMV and appeared as primary infection after1 month.Determination of HCMV pp65 IgM and HCMV p065 IgG-AI by ELISA incorporated with virus isolation and RT-PCR are helpful for distinguishing primary infections from nonpfimary infections.The detection of HCMV p65 IgM and HCMV pp65 IgG-AI by ELISA utilizing recombinant protein pp5 as antigens can be used for preliminary screening.

Objective To study the value of the liver and spleen interventional therapy in primary carcinoma of the liver with hyperspleenism. Methods The treatment was given to 25 patients. Using the method of Seldinger,the 5FRH into artery hepatica propria to infuse 2/3 of the overall dose of chemotherapy medicine and embolism was inserted into the liver segment. Then 1/3 of it was infused into artery lienalis and gelatin sponge was infused into artery lienalis Rr. lienales. Combined regimens were used in chemotherapy: 5-Fu 750mg, EADM 40mg, MMC 10mmg,and 10 of the 25 added DDP 40mg. LUF and gelatin sponge were used for embolisation. Results Of the 25 patients,the WBC and PLT increased in 25 patients 4 weeks after the operation. The difference was significant. The survival rate at 6 and 12 months was 100% and 60% respectively,with 1 case of CR,8 cases of PR and 6 cases of SD. Patients had fever after the treatment but few with abnormal liver function tests. Conclusion Liver and spleen interventional therapy may relieve hyperspleenism and suggests that the treatment of primary carcinoma of the liver is effective.

Objective To establish a new ELISA method by use of shorted recombinant antigen pp65 for detection of IgM antibodies to human cytomegalovirus (HCMV).Method According to HCMV pp65 sequence of nucleotide and amino acid, the shorted pp65 was decided by computer soft ware of protein and attained through gene engineering technique. The by computer soft ware of protein and attained through gene engineering technique. The recombinant enzyme- linked immunosorbent assay (REC-ELISA ) method was developed using pp65 recombinant protein as antigen and was applied to detecting anti-HCMV-IgM in sera. It was compared to ELISA by use of whole virus as antigen and Biocheck CMV IgM enzyme immunoassay test kit.Results To detect HCMV IgM by REC-ELISA, the best quantity of pp65 was 3.5?g percent cavity, the best dilution of HRP~*anti-HCMV IgM was 1∶[KG-*2]800 and that of serum was 1∶[KG-*2]100. The positive coefficient of variation (CV) was 9.5% in stability assay. The average CV of Inter- assay and Intra-assay was 4.5% and 9.6% respectively. The positive value of REC- ELISA was 44%, that of ELISA using whole virus was 50% and BioCheck was 45%. REC-ELISA using most suitable condition was concordant with BioCheck, which was 97.0%. Its youden′s index was 92.8% and its specificity (98.2%) was higher than that of ELISA using whole virus as antigen (90.9%), too.Conclusion REC-RLISA had good sensitivity, specificity and reproducibility. The method was easy and quick. Recombinant protein was harmless and easy gained compared with the whole virus. It can be large-scale production and standardization. Therefore, the application value and potential of REC-ELISA was very large.

Objective To detect CK-19 mRNA expression by quantitative real-time RT-PCR in axillary drainage fluid of rectal cancer and investigate its clinical significance.Methods Axillary drainage fluids were collected from 59 patients with rectal cancer and 15 patients with benign abdominal lesion from Sep.2010 to Dec.2010.Level of CK-19 mRNA in axillary drainage fluid was detected using specific primers by real-time RTPCR.The data were statistically analyzed to investigate the relationships between CK-19 mRNA level and tumor invasion,lymph node status,tumor stage and tumor differentiation level.Results The positive rate of CK-19 mRNA expression in patients with rectal cancer was 67.8％,which was significantly higher than that in patients with benign abdominal lesion.The expression of CK-19 mRNA was significantly correlated with the depth of tumor invasion,lymphnode status,tumor stage and histopathological differentiation( P ＜ 0.05 or P ＜ 0.01 ).Ck129 mRNA expression was associated with the pathological level,the higher of the lymph node translation level,the higher expression in the axillary drainage fluid after rectal cancer surgery (r =0.674,P =0.021 ).The lower of the lymph node differentiation level,the higher expression in the axillary drainage fluid after rectal cancer surgery (r =-0.741,P =0.014).Conclusion Quantitative detection of CK-19 mRNA in axillary drainage fluid of rectal cancer by RT-PCR could enhance the diagnostic sensibility of colorectal cancer micrometastases.RT-PCR assay is suitable for predicting peritoneal micrometastasis of rectal cancer,which is a reference for postoperative treatment and prognosis prediction.

Objective To explore the value of automatic photography in the observation of results of Schistosoma japoni?cum miracidium hatching experiments. Methods Some fresh S. japonicum eggs were added into cow feces,and the samples of feces were divided into a low infested experimental group and a high infested group(40 samples each group). In addition,there was a negative control group with 40 samples of cow feces without S. japonicum eggs. The conventional nylon bag S. japonicum miracidium hatching experiments were performed. The process was observed with the method of flashlight and magnifying glass combined with automatic video(automatic photography method),and,at the same time,with the naked eye observation meth?od. The results were compared. Results In the low infested group,the miracidium positive detection rates were 57.5% and 85.0%by the naked eye observation method and automatic photography method,respectively(χ2=11.723,P<0.05). In the high infested group,the positive detection rates were 97.5%and 100%by the naked eye observation method and automatic pho?tography method,respectively(χ2= 1.253,P > 0.05). In the two infested groups,the average positive detection rates were 77.5% and 92.5% by the naked eye observation method and automatic photography method,respectively(χ2 = 6.894,P <0.05). Conclusion The automatic photography can effectively improve the positive detection rate in the S. japonicum miracidi?um hatching experiments.

Objective Analysis of the complete genome sequence about the newly emerged pandemic norovirus GⅡ.4 genotype, to understand its variation characteristics.Methods 264 patients were collected with diarrhea.The RNA was extracted from 264 fe-cal specimens and cDNA was synthetized.The positive samples were amplified by PCR,the amplified fragments were sequenced. The complete genome sequences of the norovirus was sequenced and analyzed.Results A new norovirus variant strain of Jingzhou GⅡ.4,a pleiston of Sydney GⅡ.4 was isolated.A large variation was found in the new variant subtype,which was mutated inclu-ding in the hypervariable P2 domain of the major capsid protein VP1.Conclusion The result demonstrates the variant strain of Sydney GⅡ.4 was spread to China.VP1 of norovirus GⅡ.4 is evolving rapidly.The spread and evolution situation of the norovirus GⅡ.4 need to be closely monitored in China for the development of effective vaccines and therapeutic monoclonal antibodies.

Objective　To investigate the effect of congenital human cytomegalovirus（HCMV） infection on the growth of embryo and cerebral cortex. Methods　HCMV (6.0 log TCID50 in 1.0 ml/mice) was separately injected into the intraperitoneum of 10-week-old female mice on pre-mating, 3, 7, 15 gestation days. Cerebral cortices of fetuses were collected by laparotomy on 19th gestation-day just before delivery. The specimens were fixed with 4% buffered solution of paraformal dehyde. And then sectioned and stained with HE. Meanwhile, part of the specimens was used for viral isolation and HCMV DNA test by PCR. Results　In the cerebral cortex of fetal mice, it was found that the capillary was dilated and congested, and the parenchyma was infiltrated with neutrophils, monocyte for experimental group and not for control group. The degeneration, necrosis and apoptosis of neurons co-existed. The rates of virus isolation for the fetal cerebral cortex among pre-mating, 3-, 7-, 15-gestation-day groups were 60%、62%、67%、25%,respectively （χ2=13.475，P＜0.05) and the rate for 15 gestation-day group was significantly lower than others. Average weight of experimental mice for 15-gestation-day was significantly lower than control group and those for other gestation groups were not significantly different on group t-test. Conclusion　HCMV can replicate in the cerebral cortex of ICR mice, infect fetal and initiate congenital infection of cerebral cortex of fetus. In addition, embryo of different gestation stages can be infected by HCMV on experiment. HCMV delays embryo growth and causes the cerebral cortex damage.

Objective: To investigate the roots and canals morphology o f extracted maxillary first premolars. Methods: The root morphol ogy and the root canals of 115 extracted maxillary first premolars were visualiz ed on radiographs taken in the mesio-distal direction, then the teeth were cut transversely and root sections were examined, the root canal systems was analyse d with Vertucci’s classification. Results: 74(64%) teeth were w ith one root, 41(36%) with two roots, all teeth with two roots had two canals a nd each canal with one apical foramen,〔TypeⅠ(1)〕.39% of the single-root teet h demonstrated one canal, whereas 61% of the single-root teeth had two canals, 〔TypeⅡ(2-1), TypeⅣ(2) and TypeⅤ(1-2)〕.Conclusions: The roots of maxillary first premolars possesses a variety of canal system types.

Objective:To survey the continuous change features of cusp angle in anatomic artificial teeth,and to investigate their effects on occlusal balance.Methods:28 artificial teeth of a complete denture were measured by optical three-dimensional(3D)scan-ner and the corresponding digital models were constructed using the reverse engineering software Surfacer 1 0.5.The working side cusp slopes of lateroprotrusive movement including the maxillary buccal cusp distal slopes and the mandibular buccal cusp mesial slopes were studied.The straight lines from bottom to top of cusp were constructed and the corresponding projected curves in occlusal surface were generated.The angulation values of straight line and the continuous change values of projected curve were calculated.Results:The angulation values of artificial teeth cusp inclination including straight line and corresponding projected curve were obtained.The maxillary working cusp inclination varied from 1 3.7°to 25.6°,and the mandibular working cusp inclination varied from 1 1 .3°to 37.7°.Conclusion:The relatively large differences of the cusp inclination among traditional anatomic artificial teeth and the convex form of the cusp are disadvantage factors to set up stable centric stop and unhindered chew course.