Objective:To construct GM-CSF and EB virus LMP2A fusion gene recombinant adenovirus and identificate it , using recombinant adenovirus to do immunologic study.Methods:With RT-PCR and Western blot to detect GC 2A gene expression of the recombinant adenovirus , with ELISA to analysis antibodies of vAd-GC2A stimulation in murine body , lactate dehydrogenase assay to detect the mouse cellular immunity effect , thereby the recombinant adenovirus immune effect was initially analysis.T-test method was used to do preliminary analysis and evaluation of the immune effect of recombinant adenovirus .Results:GC2A fragment amplified by PCR, the size was 1 961 bp and it was the expected.The results showed, the recombinant adenovirus vAd-GC2A proteins can be recognized by sera antibody of GM-CSF ( LMP2A); the results of ELISA showed that vAd-GC2A can stimulate the generation of antibodies.EBV-positive tumor cell killing rate (%) by CTL induced recombinant adenovirus was (66.7 ±6.9)%, at the same time the adenovirus type 5 wild strains and PBS were (24.3 ±2.5)%and(32.4 ±3.1)%only(P≤0.05).Conclusion:Fusion gene has been successfully constructed , vAd-GC2A in mice have humoral and cellular immune function.The data suggest that the vAd-GC2A should play a potent role in preventing the positive EB virus tumor.

Objective To identify the expression of a fusion gene GCA formed from GM-CSF gene and LMP2A gene of Epstein-Barr virus (EBV) in a recombinant BCG (rBCG) and to study its immunoge-nicity.Methods The rBCG was constructed to express the fusion gene GCA and the expressed products were detected by Western blot assay .ELISA was performed to measure specific antibody titers in serum sam-ples from mice immunized with rBCG .Lactate dehydrogenase assay was used to analyze the cellular immuni-ty of mice.A mouse model of EBV-positive gastric carcinoma was established to evaluate the therapeutic effects of rBCG.Results The target proteins of GM-CSF and LMP2A were successfully expressed in rBCG . The specific antibodies were detected in rBCG immunized mice as indicated by ELISA .The maximum anti-body titer reached 1 ∶27 900 [(326.5±7.8) pg/ml] as injection with rBCG 5×108/mouse.The rBCG in-duced cytotoxicity of cytotoxic lymphocytes (CTLs) to EBV-positive gastric carcinoma cells (GT39) (with a killing rate of 89.6%±6.8%) was significantly higher than that of control group (P<0.05) The sizes of tumor in PBS control group [(1964.0±548.7) mm3] and BCG group [(1268.65±72.4) mm3] were big-ger than those in rBCG group [(168.64±78.80) mm3].Conclusion The rBCG expressing GM-CSF and LMP2A fusion gene was successfully constructed .The rBCG could induce humoral and cellular immune re-sponses in mice and inhibit the growth of tumor .

0.05).Conclusion GH improves FAH of children with GHD.Height at the initiation of puberty is the most significant determining factor for the long-term efficacy.Hence,it is important that the diagnosis should be made and treatment be initiated as early as possible to afford children with GHD the opportunity to make up much of their height deficit before puberty.Adequate dosage of GH should be used for the children taking initial treatment at puberty to attain satisfactory FAH.

OBJECTIVETo investigate the anti-emetic effect of ethanol extract from "WuZhuYu broth" and its mechanism.

METHODThree experiments were carried out test which extract has anti-emetic activity, such as CuSO4-induced pigeon's emetic response, gastric emptying in mice and ACh-induced or 5-HT-induced contraction in vitro gastric muscle in rats. Meanwhile, effect of anti-emetic extract on concentration-response curve to ACh, 5-HT, histamine was investigated.

RESULT50% ethanol extract and 70% ethanol extract were identified as having significantly stronger anti-emetic activities with little side effect, which showed the significant effect on concentration-response curve to ACh, 5-HT, histamine.

CONCLUSION50% ethanol extract and 70% ethanol extract contain more anti-emetic fractions, more anti-emetic fractions can be gained at the concentrations of 50% and 70% ethanol; the mechanism of anti-emetic effect is related to its antagonism to the receptors of ACh, 5-HT, histamine.

Animals ; Antiemetics ; therapeutic use ; Columbidae ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; therapeutic use ; Evodia ; chemistry ; Female ; Gastric Emptying ; drug effects ; Male ; Mice ; Muscle Contraction ; drug effects ; Phytotherapy ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Vomiting ; drug therapy

Objective To investigate the expression of osteopontin and matrix metalloproteinase-9 (MMP9),and the relationship of osteopontin and MMP9 in malignant melanoma.Methods Expression of osteopontin and MMP9 was measured by immunohistochemical SP method in 23 patients with primary cuta- neous malignant melanoma,17 patients with metastatic melanoma and 20 patients with pigmented nevus. Results Osteopontin and MMP9 were expressed respectively in 87.5% and 75.0% of 40 malignant melanoma specimens,15.0% and 10.0% of 20 pigmented nevus specimens.The expression of both osteo- pontin and MMP9 was significantly higher (both P＜0.05) in malignant melanoma than in pigmented ne- vus.There was no correlation between the expression of osteopontin and MMP9,with age,sex,lymph node metastasis or location of lesions (P＞0.05).Twenty-nine cases were positive for both osteopontin and MMP9,4 negative for either osteopontin or MMP9.Conclusion Both osteopontin and MMP9 were over- expressed in malignant melanoma,but neither was related to lymph node metastasis.

Objective To investigate the development trend of drinking-water-borne endemic fluorosis in Jiangsu Province,and to provide the basis for further prevention and treatment of the disease.Methods In 2009,eight major counties were chosen,and in each county all diseased villages were classified into light,moderate and severe disease types according to water fluorine content based on historical data,and one village was chosen from each type.In monitoring villages with improved water,one source water and three tap water samples were collected,respectively.Five water samples were collected in water unimproved monitoring villages according to water well locations of the east,the west,the south,the north and the center.The fluorine content in water was determined according to the Standard Testing Methods for Drinking Water (GB/T 5750-2006).Children aged 8 to 12 were examined for dental fluorosis by Dean method.Residents over the age of 16 were examined for clinical osteofluorosis,and two monitoring counties were chosen,then one village was respectively chosen in each county,and clinically diagnosed patients with skeletal fluorosis were examined again by X-ray.Both clinical and X-ray diagnosis were on the basis of Diagnostic Criteria of Endemic Skeletal Fluorosis (WS 192-2008).Urine samples of 30 children aged 8 to 12 and of 20 adults over the age of 16 were randomly collected and urinary fluoride was determined by F-ion selective electrode method(WS/T 89-1996).Results Of all the 24 villages of 8 counties,20 villages were water improved,and water-improvedprojects ran normally in 18 villages,while scrapped in the rest 2 villages.One hundred and two samples were tested,and the mean of water fluoride in water-improved villages was 1.10 mg/L,while in water unimproved villages and villages with water improved projects scrapped was 1.90 mg/L.The prevalence of dental fluorosis was 42.51％(854/2009).The prevalence of clinical skeletal fluorosis was 23.23％ (2024/8713) and the X-ray detection rate was 32.00％ (24/75).Six hundred and sixty-four urine samples of children were determined,and geometric mean of urinary fluorine was 1.59 mg/L,while 370 adult urine samples were determined,and geometric mean of urinary fluorine was 2.20 mg/L.Conclusions Endemic fluorosis in Jiangsu Province has not been fully controlled and there are signs of recovery.We must pay attention to water improvement measures to reduce fluoride and the management and maintenance of water improvement projects,and further strengthen the prevention and control of endemic fluorosis.

Objective To investigate HPV infection and its correlation with the expression of sur- vivin and proliferating cell nuclear antigen (PCNA) in condyloma acuminatum (CA).Methods HPV6/11 and 16/18 DNA were detected by PCR in 41 cases of CA and 23 specimens of normal skin.The expression of survivin and PCNA was measured by immunohistochemical technique in these two groups. Results The infection rates of HPV6/11 and 16/18 were 87.80% (36/41) and 19.51%(8/41),respective- ly,in CA,and 0 in normal skin epidermis.The expression of survivin and PCNA was found in 53.66% (22/41) and 87.80% (36/41) of CA ,and was 8.70%(2/23)and 47.83% (11/23) in normal skin,respec- tively.The expression of survivin was positively correlated with the high expression of PCNA in CA.The expression of survivin and PCNA was higher in HPV6/11 positive lesions than in HPV6/11 negative lesions. Conclusion HPV infection may up-regulate the expressions of survivin and PCNA,inhibit the apoptosis and promote the proliferation of keratinocytes.

OBJECTIVETo study the pathologic change and molecular regulation in cell proliferation and apoptosis of gastric mucosa in rats with chronic atrophic gastritis (CAG), and evaluate the possible mechanisms.

METHODSRats were administered with 60% alcohol or 2% salicylate sodium, 20 mmol/L deoxycholate sodium and 0.1% ammonia water to establish chronic atrophic gastritis (CAG) models. The gastric specimens were prepared for microscopic view with hematoxylin and eosin (H-E) and alcian blue (A-B) stain. The number of infiltrated inflammatory cells, the thickness of the mucosa gland layer (microm) and the number of gastric glands were calculated. The damage of barrier in mucosa with erosion or ulceration, and the thickness of mucin were examined by scanned electron microscope (SEM). The levels of PGE(2), EGF (epiderminal growth factor) and gastrin in the serum were measured with radioimmunoassay or ELISA method. The immunohistochemistry method was used to observe the number of G cells, the expression of protein of EGFR (EGF receptor), C-erbB-2, p53, p16 and bcl-2 in gastric tissue.

RESULTSUnder SEM observation, the gastric mucosa was diffused erosion or ulceration and the thickness of mucin was decreased. Compared with normal rats, the grade of inflammatory cell infiltration in CAG rats was elevated, whereas the thickness and number of gastric gland were significantly lower (P<0.05). Compared with normal level of (0.61+/-0.28) microg/L, EGF in CAG (2.24+/-0.83) microg/L was significantly higher (P<0.05). The levels of PGE(2) and gastrin in serum were significantly lower in CAG rats than that in normal rats (P<0.05). Immunohistochemistry detection showed that the number of G cell in antrum was lower in CAG group (P<0.05). Immuno-stain showed EGFR protein expression in the basal and bilateral membrane, and the cytoplasma in atrophic gastric gland, while negative expression was observed in normal gastric epithelial cells. Positive staining of p53 and p16 protein was localized in the nucleus of epithelial cells. The former was higher positively expressed in atrophic gland, while the later was higher positively stained in normal gastric tissue. bcl-2 protein was positively stained in the cytoplasma in atrophic gastric gland, while very weakly stained in normal gastric tissue.

CONCLUSIONThe pathological findings in gastric gland accorded with the Houston diagnostic criteria of antrum-predominant CAG. CAG in rats was related with the damage of barrier in gastric mucosa and the misbalance of cell proliferation and apoptosis. There was high protein expression of oncogene, while inhibitor of suppressor gene in CAG rats indicated high trend of carcinogenesis.

Animals ; Chronic Disease ; Epidermal Growth Factor ; blood ; Gastric Mucosa ; chemistry ; pathology ; Gastrins ; blood ; Gastritis, Atrophic ; metabolism ; pathology ; Immunohistochemistry ; Male ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Rats ; Rats, Sprague-Dawley ; Receptor, Epidermal Growth Factor ; analysis ; Tumor Suppressor Protein p53 ; analysis

ABSTRACT Clinical application of standards for reporting interventions in clinical trials of acupuncture (STRICTA) is introduced in this article, and improving opinions are proposed as well. STRICTA has already been extensively applied in designation of acupuncture clinical trials, composition of articles and quality assessment of acupuncture literature. According to the present version of STRICTA, it is suggested that items such as "standards and methods on acupoint selection and location", "angle and direction of needle insertion" and "whether the subjects ever have been acupunctured" should be further perfected. Individuated treat protocols which is highlighted on treatment according to differentiation of syndromes according to different opportunities and stages of diseases should be promoted so as to give better expression to the characteristics of Chinese medicine and enhance the clinical value of the relative literature.
Acupuncture Points ; Acupuncture Therapy ; methods ; standards ; Clinical Trials as Topic ; methods ; standards ; Humans ; Quality Improvement

OBJECTIVETo investigate the clinicopathologic features and immunohistochemical of the basal-like subtype of invasive breast carcinoma (BLBC), and to discuss the diagnosis standard.

METHODSImmunohistochemistry was performed in 448 cases of breast carcinoma and these cases were categorized into luminal A, luminal B, null subtypes, HER2-overexpressing and basal-like and their clinicopathologic features were observed under light microscope with stains of HE and immunohistochemical InVitrogen staining.

RESULTSAmong the breast cancer patients, the incidence of BLBC was 15.4% (69/448). Morphologic features significantly associated with BLBC constituently included nest structure and showing diffuse growth pattern, large scarring areas without cells in tumor, geographic necrosis, pushing margin of invasion, lymphocytic infiltrate in various degree in tumor stroma, syncytial tumor cell without clear boundaries, tumor cell showing vesicular unclear chromatin and nucleolus, markedly elevated mitotic count, metaplasia (all P < 0.01). Meanwhile, most BLBC showed strong immunoreactivity for CK5/6, CK14, CK17 (all P < 0.01).

CONCLUSIONBLBC showed distinct morphologic and immunophenotypic features.

Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; metabolism ; pathology ; Breast Neoplasms, Male ; metabolism ; pathology ; Carcinoma, Basal Cell ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Keratin-14 ; metabolism ; Keratin-17 ; metabolism ; Keratin-5 ; metabolism ; Keratin-6 ; metabolism ; Lymphatic Metastasis ; Male ; Middle Aged ; Receptor, ErbB-2 ; metabolism ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism