Objective To analyze the epidemiological features and influencing factors of human brucellosis in Qinghai province,and to provide scientific basis for prevention and control of brucellosis.Methods From 2006 to 2010,select the high incidence areas of brucellosis in Qinghai province and five counties(Henan,Dari,Tianjun,Ping'an and Haiyan counties) included in the “Central Subsidies to Local Public Health Special Fund Human Brucellosis Prevention and Control Projects” for the survey point,as well as high-risk employees from Qinghai Biological Pharmaceutical Factory were investigated.Combined with epidemiological questionnaire investigation [done according to the “National Human Brucellosis Surveillance Program(Trial)”],clinical symptoms and signs,confirmed human brucellosis patient were tested by intradermal allergy test,rose bengal plate agglutination test(RBPT) and standard tube agglutination test (SAT),in accordance with “Diagnostic Criteria and Principles of Management of Brucellosis” (GB 15988-1995) and“Diagnostic Criteria for Brucellosis” (WS 269-2007).Results Of 8368 serum samples detected,347 were RBPT positive,and the positive rate was 4.15％;5346 serum samples were tested by SAT,180 were positive,and the positive rate was 3.37％.In June 2009,112 employees in Qinghai Biological Pharmaceutical Factory were investigated on a follow-up survey,83 were RBPT positive,the positive rate was 74.11％; 58 were SAT positive,the positive rate was 51.79％.Eight of them were new cases and 4 were chronic brucellosis.Twenty five new cases were reported between 2006 and 2010.The peak incidence was from March to July.Most of the cases were herdsmen.ConclusionStrengthening animal quarantine,strengthening public education,and improving protection awareness,can effectively control the disease brucellosis.

Objective To study the expression of vascular endothelial growth factor(VEGF)-C, VEGF-D and their receptor-3(VEGFR-3)in patients with colorectal cancer and their clinicopathological value.Methods Eighty specimens of the colorectal cancer and thirty normal adjacent bowels were stud- ied.The expression of VEGF-C,VEGF-D and VEGFR-3 proteins and mRNAs in specimens of colorectal cancers and normal colorectal tissues was studied by Strept-avidin-biotin complex method and RT-PCR. Clinicopathological data and survival of each patient were recorded and analyzed.Results①The staining of brown or filemot in cytoplast were observed as the positive expression of VEGF-C,VEGF-D and VEGFR-3 proteins.The positive rate(48.8%,56.3%,38.8%)and mean value(1.09?1.20,1.13?1.09,0.90?1.19)of VEGF-C,VEGF-D,VEGFR-3 expressions in specimens of colorectal cancer were significantly higher than those of the normal bowel tissues(P＜0.05).The expression of VEGF- C,VEGF-D and VEGFR-3 mRNAs by RT-PCR was correlated with that of VEGF-C,VEGF-D and VEGFR-3 proteins in colorectal carcinomas and normal bowel tissues.②Significant correlation between VEGF-C(P=0.0069),VEGF-D(P=0.0024)and VEGFR-3 expression was observed in colorectal cancers;moreover,no correlation between VEGF-C and VEGF-D.③The expression of VEGF-C, VEGF-D and VEGFR-3 in colorectal cancers was not correlated with age,gender,site and dimension of lesion,types of gross and histological,degree of differentiation and liver and pulmonary metastasis,but correlated significantly with Dukes' stage(P=0.0234,P=0.0003,P=0.0429)and lymph node me tastasis(P=0.0059,P＜0.01,P=0.0068).The increased death rate(P=0.0374,P=0.0127) and poor survival(P＜0.01,P＜0.01)were observed in the colorectal cancer patients with positive ex- pression of VEGF-C and VEGFR-3 when comparing with the patients of the negative expressions,but the expression of VEGF-D in colorectal cancers was not correlated with prognosis of the patients.Con- clusions Colorectal cancer cells may secrete lymphangiogenetic growth factors VEGF-C,VEGF-D and their receptor VEGFR-3,which induce the growth of lymphatic vessel endothelium and lymphangiogene- sis by VEGF-C,VEGF-D/VEGFR-3 signaling pathway,further accelerate lymphatic metastasis of colo- rectal cancers.VEGF-C,VEGF-D and VEGFR-3 might be acted as molecular phenotypes of lym- phangiogenesis in coiorectal cancers and important markers for evaluating lymphatic metastasis and prog- nosis in patients with coloreetaI carcinoma.

We revisited the "Xiao Chaihu Decoction event (XCHDE)" occurred in late 1980s in Japan and the Bupleuri Radix related adverse drug reaction (ADR) reports in China After careful review, comparison, analysis and evaluation, we think the interstitial pneumonitis, drug induced Liver injury (DILI) and other severe adverse drug envents (ADEs) including death happened in Japan is probably results from multiple factors, including combinatory use of XCHDE with interferon, Kampo usage under modern medicine theory guidance, and use of XCHD on the basis of disease diagnosis instead of traditional Chinese syndrome complex differentiation. There are less ADE case reports related to XCHD preparation in China compared to Japan, mostly manifest with hypersensitivity responses of skin and perfuse perspiration. The symptoms of Radix Bupleuri injection related ADEs mainly manifest hypersensitivity-like response, 2 cases of intravenous infusion instead of intramuscular injection developed hypokalemia and renal failure. One case died from severe hypersensitivity shock. In Chinese literatures, there is no report of the interstitial pneumonitis and DILI associated with XCHDG in Japan. So far, there is no voluntary monitoring data and large sample clinical research data available. The author elaborated the classification of "reevaluation" and clarified "re-evaluation upon events" included the reaction to the suspected safety and efficacy events. Based on the current status of the clinical research on the Radix Bupleuri preparations, the author points out that post-marketing "re-evaluation upon suspected event" is not only a necessity of continuous evaluation of the safety, efficacy of drugs, it is also a necessity for providing objective clinical research data to share with the international and domestic drug administrations in the risk-benefit evaluation. It is also the unavoidable pathway to culture and push the excellent species and famous brands of TCM to the international market, in that it enhances the pharmaceutical manufactures the ability to control enterprise risk and eliminate the negative impact on traditional Chinese medicine. With the widely used and ADR report concentrated Radix Bupleuri injection as an example, the author provide a new approach to post-marketing clinical research.
Bupleurum ; chemistry ; Drug Compounding ; Drug-Related Side Effects and Adverse Reactions ; etiology ; Drugs, Chinese Herbal ; adverse effects ; Humans ; Product Surveillance, Postmarketing ; methods ; Safety

Electromagnetic Fields ; Magnetic Fields ; Microwaves ; adverse effects ; Television

OBJECTIVETo demonstrate the changes of PAI-1 activity and mRNA expressions induced by tumor necrosis factor alpha (TNF-alpha) or minimally modified low-density lipoprotein (mmLDL) in human umbilical vein endothelial cells (HUVECs).

METHODSThe PAI-1 activity in HUVECs culture medium was measured by chromogenic assay. The PAI-1 mRNA expressions were determined by Northern blot. The mitogen-activated protein kinase-kinase (MAPKK) inhibitor and protein kinase C (PKC) inhibitor were used to intervene the inductions by TNF-alpha or mmLDL.

RESULTSThe PAI-1 activity and mRNA level were increased when HUVECs were exposed to TNF-alpha or mmLDL. The MAPKK inhibitor (PD98059, 60 mumol/L) could markedly inhibit PAI-1 activity and mRNA expression induced by TNF-alpha (100 U/ml) or mmLDL (50 micrograms/ml). But Staurosporine (10 nmol/L) and H7 (15 mumol/L), which were the PKC inhibitors, had no significant inhibition effect.

CONCLUSIONS(1) TNF-alpha or mmLDL could induce PAI-1 activity and mRNA expression in HUVECs. (2) Increase of PAI-1 activity induced by TNF-alpha and mmLDL was related to its mRNA expression. (3) The MAPK pathway may play a role in PAI-1 expression induced by TNF-alpha or mmLDL.

Endothelium, Vascular ; cytology ; metabolism ; Female ; Humans ; Lipoproteins, LDL ; pharmacology ; Mitogen-Activated Protein Kinases ; metabolism ; Plasminogen Activator Inhibitor 1 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Tumor Necrosis Factor-alpha ; pharmacology ; Umbilical Veins ; cytology

ObjectiveTo survey 0—6 years old disabled children in Guiyang.Methods4 999 children aged 0—6 years old chosen randomly from 9 145 children of 12 blocks were investigated with the disability sampling survey criteria of 1987.ResultsThe rate of disabled children in Guiyang was 1.58%, and was not significantly different compared with that of whole country (1.36%) and other three areas including Siping City (P>0.05),but significantly higher than that of other six districts including Heping district of Tianjin municipality (P<0.05). All kinds of disability found by this survey were separately intellectual disability (0.92%),comprehensive disability (0.28%),limb disability (0.24%),vision disability (0.10%),psychiatric disability (0.03%),and hearing disability (0.02%). Rates of intellectual and vision disabilities were significantly higher than that of whole country ( P>0.05).ConclusionSome diseases are directly relative to occurrence of disability. The poor income of the children family,lower education of his parents and not gotten preschool education were risk factors of disability. The institution based rehabilitation and community based rehabilitation are main rehabilitation forms to disabled children.

Comparison studies of recombinant prourokinase(pro-UK) in various host cells,and expression vectors were carried out. Expression levels of vectors constructed in this study in different cell lines were compared. Mammalian cells expressing pro-UK were established. The levels of pro-UK expression in recombinant Namalwa, Vero and Sp2/0 cells are 200,12.5 and 50 IU/( 106 cells 24h). pro-UK purities separated by immunoaffinity chromatograph are above 90 %. Immunoabsorbent assay showed the ratio of pro-UK in CHO cells is lower than that from Vero and Namalwa cells. This study provide new host cells for pro-UK production.

OBJECTIVETo evaluate the clinical significance of detection on lymphatic microvessel, lymphatic microvessel density (LMVD) and vascular endothelial growth factor-C (VEGF-C) in patients with colorectal carcinoma.

METHODSEighty tissue specimens of the colorectal carcinoma and the peritumoral tissue and thirty of adjacent normal bowel tissue were collected. The lymphatic microvessel and LMVD were determined by 5'-nucleotidase histochemical staining. The expression of VEGF-C protein and VEGF-C mRNA in specimens of colorectal carcinoma and normal colorectal tissues were studied by RT-PCR and immunohistochemical methods utilizing strept-avidin-biotin complex. Clinicopathological data and survival of each patient were obtained and analyzed.

RESULTS(1) The brown or filemot stained lymphatic microvessels were observed in specimens from the colorectal carcinoma, the peritumoral tissue and the normal bowel. Collapsed, nonfunctional lymphatic vessels were observed in the intratumoral tissue, and plenty of lymphatic vessels with large lumen referred as functional lymphatic vessels were observed in the peritumoral tissue. (2) The mean value of LMVD in the peritumoral tissue was significantly higher than that in the normal bowel tissue (9.76+/-2.85 vs. 5.49+/-1.43, t=8.220, P<0.01) and tumor tissue (9.76+/-2.85 vs. 2.13+/-0.96, t=15.118, P<0.001). (3) The positive rate (48.8% vs. 0, P<0.01) and mean value (1.09+/-1.20 vs. 0, P<0.01) of the VEGF-C protein expression in colorectal carcinoma specimens were significantly higher than that of the normal bowel tissue. The expression of VEGF-C protein was consistent with the expression of VEGF-C mRNA. The VEGF-C expression in intratumoral tissue demonstrated significant correlation with LMVD in the peritumoral tissue of colorectal carcinoma. (4) Both LMVD in the peritumoral tissue and the expression of VEGF-C in the intratumoral tissue correlated significantly with Dukes' stage (P<0.0001 and P=0.0234), lymph node metastasis (P<0.0001 and P=0.0059), and survival (P<0.0001 and P<0.0001), but not with age, sex, location and dimension of lesion, gross and histological type. Also, there was a positive significant correlation of LMVD in the peritumoral tissue with degree of differentiation (P=0.0168) and metastasis to the liver or the lung (P=0.0088).

CONCLUSIONSLymphatic microvessels in the peritumoral tissue are functional. The functional lymphatic microvessels, increased LMVD in the peritumoral tissue and the expression of VEGF-C in the intratumoral tissue may act as the morphological features and the molecular phenotype of lymphangiogenesis in colorectal carcinoma, and also as important predictive markers for evaluating lymphatic metastasis and prognosis in patients with colorectal carcinoma.

Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; metabolism ; pathology ; Female ; Humans ; Lymphangiogenesis ; Lymphatic Metastasis ; Lymphatic Vessels ; pathology ; Male ; Microvessels ; Middle Aged ; Neoplasm Staging ; Prognosis ; Vascular Endothelial Growth Factor C ; metabolism ; Young Adult

OBJECTIVETo study lymph node micrometastases (LNMM), expression of nm23-H(1), MMP(9), TIMP(2) proteins, and their relationship and clinical significance in patients with stage Dukes B colorectal cancer.

METHODSThirty patients with stage Dukes B colorectal cancer were studied. LNMM in these patients was detected by immunohistochemical anti-cytokeratin 20 (CK20) staining. The expression of nm23-H(1), MMP(9) and TIMP(2) proteins in primary tumors was examined by Strept-avidin-biotin complex method. Clinical-pathological data and survival of each patient were recorded and analyzed.

RESULTS(1) The positive dyeing of CK20 was observed in 26.7% for cases and in 7.8% for lymph nodes of 30 patients with stage Dukes B colorectal cancer. (2) Different expression of nm23-H(1) and MMP(9) proteins in the patients between stage Dukes B and stage Dukes CD was observed (P < 0.05). The decreased nm23-H(1) expression, and/or the increased MMP(9) expression in primary stage Dukes B tumors were significantly associated with LNMM (P < 0.05). Sensitivity and specificity for detection of LNMM by using nm23-H(1) or MMP(9) were respectively 62.5% and 81.8% or 75.0% and 69.8%. If by combining nm23-H(1) with MMP(9), specificity for detection of LNMM became 90.9%. The expression of TIMP(2) protein was not related with stage Dukes and LNMM. (3) The percent of tumor recurrence and/or metastasis for the stage Dukes B patients with LNMM was significantly higher than that for the patients without LNMM (P < 0.05), but the survival percent for the patients with LNMM was significantly lower than that for the patients without LNMM. The outcome for the patients with nm23-H(1) (-) LNMM (+) or MMP(9) (+) LNMM (+) was significantly worse than that for patients with nm23-H(1) (+) LNMM (-) or MMP(9) (+) LNMM (-) (P < 0.05).

CONCLUSIONSLNMM is detected by immunohistochemical anti-CK20 staining. The expression of nm23-H(1) and MMP(9) in primary stage Dukes B tumors was significantly associated with LNMM. The outcome in the LNMM patients with nm23-H(1) (-) and/or MMP(9) (+) were worse. Combining examination of CK20 for lymph nodes with expression of nm23-H(1) and MMP(9) for primary tumors is of important clinical significance for staging of Dukes, selection of adjuvant treatment and evaluation of prognosis in patients with colorectal cancer.

Colorectal Neoplasms ; metabolism ; pathology ; therapy ; Humans ; Keratins ; metabolism ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Matrix Metalloproteinase 9 ; metabolism ; NM23 Nucleoside Diphosphate Kinases ; Neoplasm Staging ; Nucleoside-Diphosphate Kinase ; metabolism ; Prognosis ; Tissue Inhibitor of Metalloproteinases ; metabolism

Danggui, Agelicae Sinensis Radix, is a widely used Chinese herb to enrich blood, but its quality cannot be effectively assessed by the known chemical markers such as ferulic acid, ligustilide, polysaccharides, etc. A new bioassay was therefore developed to quantify the Enrich-Blood Bioactivity (EBB) for the quality assessment of Danggui raw materials. Danggui sample was first extracted with ethanol and water, respectively. Then the ethanolic extract and water extract were mixed as a test sample to quantify the amount of EBB by mice experiment. The blood deficiency mode in mice was developed by intraperitoneal injecting cyclophospharmide and phenylhdrazine hydrochloride. The quantity of red blood cell was chosen as EBB marker. Cyclosporine A was chosen as a control substance. EBB in analytes was quantified by the amount reaction of parallel line analysis (3, 3') method. The results indicated that the reliability test for quantifying EBB was passed through and the measured value was valid. The analytes showed the significant EBB (P < 0.05). The correlation coefficient was 0.9984 (n=5) between the amount of cyclosporine A (0.035-0.56 g x kg(-1)) and the increased number of red blood cell. The relative standard deviation (RSY) on the amount of EBB was estimated to be 6.15% (n = 6) by six replicated tests, and the confidence limit rate was 26.68% (n = 6). Five Danggui samples, which were collected from different cultivation areas with various morphological characters, showed the variety of EBB in the range of 21.95-44.16 U x g(-1). It is concluded that the developed method is accurate to quantify the EBB of Danggui raw materials, and is therefore suitable to assess its quality.
Angelica sinensis ; chemistry ; Animals ; Biological Assay ; methods ; Drugs, Chinese Herbal ; pharmacology ; Erythrocyte Count ; Erythrocytes ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; Plant Roots ; chemistry