Objective:To assess the efficacy of time-resolved fluoroimmuno assay(TRFIA) for quantative determination of?-fetoprotein(AFP) in comparison with radio immuno assay(RIA) and enzme linked immunoabsorbent assay(ELISA). Methods:Thirty serum samples from randomly selected patients were collected to test AFP content with TRFIA,RIA and ELISA respectively.Results:The linear range of AFP was 1 to l 000 ng/ml for TRFIA,5～400ng/ml for RIA and 5～300ng/ml for ELISA.The intraassay coefficients of variation(CV) and interassay CV were 3.3% and 4.7% for TRFIA,5.3% and 8.9% for RIA,and 10.5% and 15.2% for ELISA,respectively.Correlation tests revealed that the results of RIA were more closely correlated with those of TRFIA than with those of ELISA.Conclusion:TRFIA is shown to be a better method for AFP quantitation than RIA and ELISA in terms of precision and linear range.

Objective:To discuss the rule of the prescriptions of professor JIANG Wen-zhao in treating the spleen and stomach ailment. Methods:Use the frequency analysis to analyze the 98 pieces of prescriptions of professor JIANG Wen-zhao in treating the spleen and stomach ailment. Results:To understand medicine state of professor JIANG Wen-zhao in treating the spleen and stomach ailment preliminarily,select the main medicine in treating diseases of the spleen and stomach,and make clear the significance of these medicines to the corresponding diseases. Conclusion:The basic rules of professor JIANG Wen-zhao in treating the spleen and stomach disease are regulating qi,removing dampness and reinforcing asthenia. Qi regulating herbs,diuretics for eliminating dampness,tonifying deficiency herbs,dampness-removing herbs,blood-activating and stasis-removing herbs,heat-clearing herbs are the key herbs.

[Summary] This article reviewed the clinical features in two diabetes cases complicated with nasal‐brain mucormycosis by pathology and pulmonary mucormycosis. In one patient with diabetes combined with pulmonary mucormycosis ,manifestations of chest pain and hemoptysis ,the right upper lung biopsy tip morphology was consistent with Mucor ,successfully cured with amphotericin and operation treatment. The other patient with diabetes‐complicated nasal mucormycosis with headache and nasal cavity clump , showed black caseous matter ,finally hemiplegia. A nasal cavity tumor was proved by pathological examination. After treatment with fluconazole and operation ,the patientdied. Mucormycosisis rare and may be secondary to diabetes and hypoimmunity disease. It needs rapid diagnosis and treatment.

Objective To explore the clinical expression of P53, Livin and PARP in the epithelial ovarian cancer and its correlation with the chemotherapy resistance and clinical prognosis.Methods 74 specimen of epithelial ovarian cancer confirmed from January 2009 to June 2011 in our gynecology department were selected.During the follow-up visit, the subjects were divided into chemotherapy sensitivity group and chemotherapy resistance group according to the recurrence cases, the clinical expression and survival rate for two groups were compared, the influence factors of survival time were analyzed.Results The positive rate of P53, Livin and PARP for chemotherapy sensitivity group was 47.1%, 56.9%and 52.9%;the positive rate for chemotherapy resistance group was 73.9%, 95.7% and 95.7%,the diyforences were significant(P<0.05).After 1, 3 and 5 years of treatment, the survival rate for chemotherapy sensitivity group was 100.0%, 82.4% and 66.7%,The survival rate for chemotherapy resistance group was 87.0%, 26.1% and 8.7%,the diyforences were significant(P<0.05).Based on the Cox regression model, the influence factors of the patient's age, pathological differentiation degree, clinical staging and chemotherapy sensitivity were introduced.It was known that the patient's survival time was greatly influenced by clinical staging and chemotherapy sensitivity (P<0.05).Conclusion For patients with epithelial ovarian cancer, the expression of P53, Livin and PARP is correlated with chemotherapy resistance.Therefore, the clinical effect is predictable, for patients with higher expression, the personalized therapy can improve the patient's prognosis.

Objective To study the features of esophageal motility in the cases with myasthenia gravis(MG).Methods 15 healthy controls and 45 patients with MG were tested by using a low-compliance four-lumen hydraulic infusion system,respectively.Focused on the following parameters:PP,UESP,UEP,MEP and MERP.Results There were significant differences of the parameters in between MG cases and healthy controls PP (73.24 ± 31.40) mmHg (1 mmHg =0.133 kPa) vs.(103.78±29.47) mmHg,P=0.002;UESP(41.75 ±21.04) mmHg vs.(60.59 ±17.97) mmHg,P=0.003;UEP (56.63 ±30.26) mmHg vs.(78.98 ±30.14) mmHg,P =0.016;MEP(53.96 ±23.25) mmHg vs.(75.11 ±23.75) mmHg(P =0.004).However,MERP of MG cases or healthy controls seemed to be similar[(-7.76 ± 5.94) mmHg vs.(-7.58 ± 5.76) mmHg,P =0.91).Additionally,the above-mentioned parameters in the cases with generalized myasthenia gravis or dysphagia were significantly different compared with other subtypes or healthy controls (P ＜ 0.01).However,there were no significant differences of the parameters in between ocular MG and healthy controls,or in between MG with and without thymoma.Conclusion The upper and middle part of esophageal motility dysfunction is very common in MG cases,especially in those with dysphagia or generalized MG,characterizing by the declined pressure.Manometry in MG cases can help us classify the subtypes of MG and verify their esophageal motility functions.

AIM: To explore the effect of collagen and C-erbB-2 protein on the adhesion and the proliferation in hepatocellular carcinoma cells.METHODS: Hepatocellular carcinoma cell line(HepG-2) identified to positive for C-erbB-2 gene was used to study the adhesion and the growth feature by the action of rat tail collagen and C-erbB-2 antibody.RESULTS: The action of rat tail collagen to potentiated the adhesion in HepG-2 cells was significantly but no proliferation effect was observed. C-erbB-2 antibody inhibited the adhesion and proliferation of HepG-2 cells and also abolished the potentiated effect of rat tail collagen on the adhesion in HepG-2 cells.CONCLUSIONS: The signaling transduction mediated by C-erbB-2 protein was correlated to the adhesion and the proliferation of HepG-2. The blockage of C-erbB-2 gene signal transduction may be a strategic target to the treatment of liver cancer in the future.

AIM: To examine the effect of insulin on the proliferation of human hepatocellular carcinoma cell and its possible mechanisms. METHODS: The human hepatocellular carcinoma cell line(HepG-2) was used to study the changes in calcium ion across plasma membrane (Ca 2+ APM)under the action of insulin by the assay of atomic absorption spectrum, and in the proliferation under the action of insulin and calcium ion antagonist (isoptin). RESULTS: The influx of Ca 2+ APM and the proliferation was increased after insulin administration, but the proliferation was inhibited by isoptin. CONCLUSION: Changes in the homeostasis of calcium ion across plasma membrane was involved in the effect of insulin on the proliferation of human hepatocellular carcinoma.

AIM: To explore the effects of PMA(phorbol-12-myristate-13-acetate,a tumor promoter, mimicking the action of diacylglycerol on PKC)and laminin on the adhesion and the proliferation of human hepatocellular carcinoma cells, and provide a new clue to liver cancer treatment. METHODS: Human hepatocellular carcinoma cell line(BEL-7402)was used to identify the endogenous laminin and protein kinase C-?(PKC-?) expression, and the effects of laminin and PMA on the adhesion and the proliferation were also investigated in vitro. RESULTS: By the effect of exogenous laminin, human hepatocellular carcinoma cell (BEL-7402) possessed endogenous laminin expression and increased the adhesion and the proliferation, which was showed the synergistic action by the effect of PMA in combination. By the action of PMA alone,the proliferation and the PKC-? expression increased by exogenous laminin were decreased, and the adhesion and the endogenous laminin expression were increased. CONCLUSIONS: The finding suggested that the adhesion and the proliferation of human hepatocellular carcinoma cell were closely related to the effects of endogenous or exogenous laminin ,which were associated with cPKC-? activity. Therefore,the application of anti-laminin antibody in combination with PKC antagonist might be a new clue to find out the therapy for liver cancer.

Objective To investigate the human papillomavirus 16/18 (HPV16/18) infective status and its relationship with the expression of p16,CK17,Ki-67 in immature squamous metaplasia(IM),cervical intraepithelial neoplasia (CIN) and cervical carcinomas (CC).Methods In situ hybridization (ISH) and immunohistochemistry were applied to detect the expression of HPV16/18,p16,CK17,Ki-67 on tissues from 30 IM,60 CIN and 30 CC.Results The expression rates of HPV16/18 in LSIL,HSIL and CC were 30 ％ (9/30),60 ％ (18/30) and 77 ％ (23/30),higher than that in IM [6.7 ％ (2/30)].HPV16/18 was positively expressed with the progression of cervical squamous epithelial lesions.The expression rates of CK17 in IM was 90 ％ (27/30),higher than that in LSIL,HSIL and CC [17 ％ (5/30),10 ％ (3/30),6.7 ％ (2/30)].The expression rates of p16,Ki-67 were 83 ％ (25/30) and 87 ％ (26/30) in LSIL,90 ％ (27/30) and 93 ％ (29/30) in HSIL,97 ％ (29/30) and 97 ％ (29/30) in CC,higher than that in IM [13 ％ (4/30) and 10 ％ (3/30)].The expression of p16,Ki-67 were particularly seen in HPV16/18-positive HSIL and CC,and the correlation were observed.Conclusion HPV16/18 infection is highly associated with the degree of squamous intraepithelial neoplasia and carcinomas of cervix and upregulated p16 and Ki-67,which suggested that HPV16/18 maybe cause mutation of p16 gene.With the progression of cervical squamous epithelial lesions,positive expression rates of CK17 were decreased compared with increased of HPV16/18,p16,Ki-67.Combined detection of HPV16/18,p16,Ki-67 is helpful for diagnosis and classification of cervical squamous epithelial lesions.

Objective To establish differential protein expressing profiles of human gastric adenocarcinoma cell in primary or metastatic lymph node tissues by two-dimensional differential gel electrophorosis (2-D DIGE) so as to investigate the metastatic molecular mechanism of the gastric cancer.Methods After obtaining 6 samples of human primary gastric cancer and metastatic lymph node tissues,with manual no-staining frozen sections microdissection,human gastric adenocarcinoma cells from primary or metastatic lymph node tissues were isolated,and then the total proteins were extracted and purified.Highly sensitive 2-D DIGE was used to separate the total protein differentially expressed in the cells.The proteins were visualized by using a fluorescence scanner at appropriate wavelengths for Cy2,Cy3 and Cy5 dyes (Typhoon 9400).Image analysis was carried out with the DeCyder-Differential analysis software (Biological Variation Analysis version 5.0).Results Not only can the study procure defined adenocarcinoma cell populations from gastric primary or metastatic lymph node tissues,but also can resolve the problem of the change in 2-D DIGE patterns because of the varying in protein changes owing to dyeing.All these showed that the technique was simple,easy to perform,versatile and of particular usefulness when laser capture microdissection (LCM) was practically unavailable.The 2-D DIGE patterns with high resolution and reproducibility from adenocarcinoma cells in gastric primary or metastatic lymph node tissues were obtained.The number of spots in Gel1,Gel2 were 1 416 (similar 1 062,decrease 277,increase 77),1 299 (similar 1 050,decrease 157,increase 92),respectively.A total of 11 differential proteins were acquired by image analysis with DeCyder-Differential analysis software (Biological Variation Analysis version 5.0).Conclusions In this report,a simple,easy to proform method of protein epuration,manual no-staining frozen sections microdissection is described,and have used the highly sensitive 2-D DIGE for the identification of proteins differentially expressing in human gastric adenocarcinoma cells from primary or metastatic lymph node tissues.These results provide a fundamental basis for further study of metastatic mechanism of gastric cancer and screen its specific markers.