Objective To obtain the fusion genes of several human orphan G protein coupled receptors (oGPCRs) with Gi1α subtype of G protein and their expression system. Methods The whole open reading frames of GPR45, GPR85, GPR174 and Gilα were cloned by RT-PCR from HepG2 cDNA separately,and the corresponding fusion genes were amplified by overlap extension PCR. Then, the fusion genes-containing pBacmids were successfully constructed with the Bac-to-Bac baculovirus expression system indicated by specific transposition and virus recombination. The insect Sf9 cells were transfected with pBacmid-oGPCRs-Gi1α, and the supernatant containing recombinant virus was harvested. With the supernatant, insect Sf9 cells were infected under an optimized condition (MOI=5, infection time=72 h) and the fusion proteins were prepared and detected by Western blotting.Results The three fusion genes of GPCR45, GPR85 or GPR174 with Gi1α were obtained. The corresponding fusion proteins could be properly prepared in Sf9 cells.Conclusion Human oGPCRs could be fused with Gilα, and the fusion genes could be expressed using the Bac-to-Bac baculovirus expression system in insect Sf9 cells.

The leading cause of drug withdrawal from market and clinical trials failure is drug-induced liver injury (DILI). Varying clinical, histological and laboratory features of DILI, as well as undefined underlying mechanisms, hinder patients to be diagnosed in the early-stage of the disease and receive effective treatments. Conventional indicators, like serum transaminases and bilirubin, have inevitable limitations referring to sensitive prediction and specific detection of DILI. In order to reduce the occurrence of DILI, researchers have attempted to discover potential biomarkers with higher specificity and sensitivity from blood and urine in recent years. This article aims to review recent advances in biomarkers of DILI.
Biomarkers ; blood ; urine ; Chemical and Drug Induced Liver Injury ; diagnosis ; Humans ; Sensitivity and Specificity

Objective To investigate the clinicopathology and immunophenotype of primary non- Hodgkin lymphoma(NHL)of the female genital system,and to analyze the prognosis of such tumors. Methods Clinicopathologic features of 43 cases of primary NHL of the female genital system were studied retrospectively,with the histological classification based on the Classification of Haematopoietic and Lymphoid Tumors(WHO,2001).Immunochemistry technique,in-situ-hybridization and polymerase chain reaction methods were used to detect the immunophenotype,epstein barrvirus(EB)virus infection status and immunoglobulin heavy chain gene rearrangement,respectively.Results(1)Primary lesions:there were 24 cases of lymphoma originating in the ovary,3 cases in the endometrium,10 cases in the cervix,2 cases in the vagina and 4 cases in the vulva.(2)Staging:12 cases(28%)were in stage Ⅰ,9 cases (21%)in stage Ⅱ,and 22 cases(51%)in stage Ⅲ.(3)Histological classification:37 cases(86%)were diffuse large B cell lymphoma(DLBCL),3 cases were Burkitt lymphoma and the remaining 3 cases were unspecified peripheral T-cell lymphoma according to biopsy,immunophenotype analysis,in-situ- hybridization technique and IgH gene rearrangement detection.(4)Prognosis analysis:increase in the level of lactic acid dehydrogenase,stage Ⅲ,DLBCL and single operation suggest poor prognosis.Conclusions Establishment of the diagnosis of primary NHL of the female genital system is based on biopsy, immunophenotype analysis,in-situ-hybridization technique and IgH gene rearrangement detection,which play important roles in diagnosis and differential diagnosis of the tumor.Combined therapy is the first choice of therapeutic regimens.

Objective To understand the current situation of iodine deficient disorder(IDD) 10 years after achieving the stage goal of eliminating IDD in Longyan city and to evaluate the effect of prevention and treatment measures, and to provide the basis for the development of control strategies. Methods There were 7 counties in the city, and each county(city, district) was as a unit to carry out the inspection for organization and leadership,iodine salt management, monitoring and control, health education (referred to as the four management indicators)according to "The County-Level Assessment and Evaluation Implementation Detailed Rules of Realizing the Goal to Eliminate IDD in Fujian Province". According to the east, west, south, north and middle positions in each county,a village and a primary school were selected. Forty 8 to 10 year-old students in each school were randomly selected to check thyroid and among them 20 students were collected urine samples to determine urinary iodine. Nine townships were selected in the 7 counties of the city and among which 4 administrative villages were selected in each township. Eight edible salt samples from each household in each administrative village were collected to test salt iodine. Goiter was examined by palpation, the level of urinary iodine was examined by arsenic and cerium spectrophotometry, salt iodine was detected by direct titration. Results The average score of the four management indicators was 94.1 in Longyan city. The adjusted goiter rate of children aged 8 - 10 years old was 1.9%. The median of urinary iodine was 278.6 μg/L, among which less than 100 μg/L accounted for 4.57%(32/700), 100 -＜ 200 μg/L accounted for 24.00%(168/700), 200 - ＜ 300 μg/L accounted for 25.29%(177/700), and higher than 300 μg/L accounted for 46.14%(323/700). The using rate of qualified iodized salt was 98.86%. The coverage rate of iodized salt was 99.50%, the qualified rate of iodized salt was 99.35%, and the rate of non-iodized salt was 0.50%. All the indicators had reached the national standard to eliminate IDD. Conclusions After achieving the stage goal of eliminating IDD, the disease is stable and the effect of control measures are significantly. But the iodine provided has a trend of more than suitable. Therefore, it is reasonable to reduce the current salt iodine content.

Thrombospondin-1,an important extracellular matrix glycoprotein,is recognized as an effective endogenous angiogenesis inhibitor and can affect and regulate the adhesion,motility and proliferation of endothelial cells,as well as closely correlated with the development and progress of tumor and neovascular diseases.In recent years,it has been found that Tsp-1 is associated with many pathologic changes in diabetic retinopathy and is critical for the maintenance of retinal vascular balance.And the mechanism of action of TSP-1 and its role in the development of DR will be reviewed in this paper.

Worldwide sales of biologic drugs exceeded 100 billion USD in 2011. About 32% is from therapeutic monoclonal antibody (mAb). With many blockbuster biopharmaceutical patents expiring over the next decade, there is a great opportunity for biosimilar to enter the worldwide especially emerging market. Both European Medicines Agency (EMA) and Food and Drug Administration (FDA) have introduced regulatory frameworks for the potential approval of biosimilar mAb therapeutics. Rather than providing a highly abbreviated path, as in the case for small molecule chemical drug, approval for biosimilar mAb will require clinical trial and the details will be very much on a case-by-case basis. Since mAb is the dominant category of biologic drugs, mAb will be the focus of this review. First, the United States (US) and European Union (EU) approved mAb and those in phase 3 trials will be reviewed, then strategies on how to win biosimilar competition will be reviewed.
Animals ; Antibodies, Bispecific ; therapeutic use ; Antibodies, Monoclonal ; therapeutic use ; Antibodies, Monoclonal, Humanized ; therapeutic use ; Biosimilar Pharmaceuticals ; standards ; Clinical Trials, Phase III as Topic ; Drug Approval ; European Union ; Humans ; United States ; United States Food and Drug Administration

OBJECTIVETo observe the clinical efficacy on pain in postmenopausal osteoporosis treated with the warm acupuncture therapy and discuss its effect mechanism.

METHODSNinety cases of postmenopausal osteoporosis were randomized into a warm acupuncture group, an electroacupuncture group and a medication group, 30 cases in each group. In the warm acupuncture group and the electroacupuncture group, Dazhu (BL 11), Shenshu (BL 23) and Xuanzhong (GB 39) were selected bilaterally and stimulated with the warm acupuncture and electroacupuncture therapies respectively, once a day for 30 days totally. In the medication group, caltrate-D tablets were prescribed, 600 mg, once a day for 30 days totally. The changes in the bone density T value, visual analogue scale (VAS) score, serum insulin like growth factor 1 (IGF-1), interleukin 6 (IL-6) and tumor necrosis factor (TNF-alpha) were observed before and after treatment in the three groups.

RESULTS(1) The bone density T value in the patients of postmenopausal osteoporosis did not change obviously after 30 days treatment with the three therapies; (2) VAS score was all reduced after treatment, in which, the result in the warm acupuncture group was the most obvious (6.73 +/- 0.24 before treatment vs 4.43 +/- 0.26 after treatment). The value after treatment in the warm acupuncture group was different significantly as compared with the electroacupuncture group (5.13 +/- 0.31) and the medication group (5.17 +/- 0.33, both P < 0.05). (3) The level of serum IGF-1 was improved after treatment in the warm acupuncture therapy [(119.5 +/- 20.1) ng/mL before treatment vs (156.5 +/- 23.9) ng/mL after treatment], which was more apparent as compared with the electroacupuncture group [(136.3 +/- 24.5) ng/mL] and the medication group [(127.7 +/- 22.1) ng/mL, all P < 0.05]. Concerning to reducing the levels of IL-6 and TNF-alpha in serum, the results in the warm acupuncture group were superior to the other two groups (all P < 0.05).

CONCLUSIONThe warm acupuncture therapy achieves the significant efficacy on pain in postmenopausal osteoporosis, which could be related to increasing the level of IGF-1, decreasing the levels of IL-6 and TNF-alpha, promoting bone formation and inhibting bone absorption.

Acupuncture Therapy ; methods ; Bone Density ; Female ; Humans ; Insulin-Like Growth Factor I ; metabolism ; Interleukin-6 ; blood ; Middle Aged ; Osteoporosis, Postmenopausal ; blood ; physiopathology ; therapy ; Pain Management ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo study the stability of chlorogenic acid, cynaroside and 3,5-O-discaffeoylquinc acid in Hangbaiju and Gongju and to predict their term of validity.

METHODHangbaiju and Gongju were incubated in an environmental chamber at different temperatures and relative humidities. After the incubation, quantitative determination of chlorogenic acid, cynaroside, 3,5-O-discaffeoylquinc acid in Hangbaiju and Gongju were measured by HPLC. The effective period of the preparation was calculated according to Arrhinius index law. Quantitative determination of 3,5-O-discaffeoylquinc acid, cynaroside, chlorogenic acid in Hangju and Gongju were analyzed by HPLC.

RESULTThe stable life of Hangbaiju has been determined as 2.25 years. The stable life of Gongju has been determined as 4.31 years.

CONCLUSIONThe high temperature is not conducive to the stability of Hangbaiju and Gongju, which needs to be placed in a dark and cool place.

Chlorogenic Acid ; analysis ; Chromatography, High Pressure Liquid ; Chrysanthemum ; chemistry ; Drug Stability ; Drugs, Chinese Herbal ; chemistry ; Temperature

Objective To understand the current situation of iodine deficiency diserders(IDD) in Longyan City and to evaluate the effect of prevention and control measures of IDD in order to provide evidence for formulating prevention and control tactics. Methods During the year of 2006 and 2007, the 30 primary schools were screened by population proportion survey(PPS) from the 7 counties of Longyan City. Forty children aged 8-10 years in each school were randomly selected as a group to examine thyroid, and 7 children in each group were selected to measure the urine iodine and the salt iodine at the same time. The goiter rote, the median urinary iodine, the consumption rate of qualified iodized salt, the iodine salt coverage rate, the rate of qualified iodized salt and the non-iodized rate were detected. Results The goiter rate of children aged 8-10 years old in Longyan City was 0.94%(79/8438). The median urinary iodine was 259.12 μg/L. The consumption rate of qualified iodized salt was 97.86% (1462/1494). The iodine salt coverage rate was 99.46%(1486/1494). The rate of qualified iodized salt was 98.38 (1462/1486), and the non-iodized rate was 0.54% (8/1494). Conclusions All indicators have reached the national standard of eliminating IDD in Longyan City.

Objective To investigate whether mesenchymal stem cells can promote the recovery of acute renal tubular damage induced by mercuric chloride and to explore its possible mechanism.Methods Acute renal failure rat model was established by intraperitoneal injection of mercuric chloride.SD rats were randomly divided into three groups which were MSCs injection group, saline infusion group and normal control group.Seven days later,the changes of rat weight,survival,renal function and pathology were observed;PCNA,ED-1 and GFP were detected by immunohistochemistry; The expression of cytokines in kidney and the distribution of GFP plasmid-transfected MSCs in kidney were examined by RT-PCR.Results MSCs infusion ameliorated the decline of rat weight,survival, renal function,and pathological changes.PCNA and ED-1 positive cells in MSCs group were fewer than those in saline group.Expression of growth factors EGF,PDGF,HGF were obviously up- regulated and pre-inflammatory cytokines TNF-?was significantly reduced in MSCs-treated kidneys. GFP-labelled MSCs occurred occasionally in renal interstitium of MSCs-treated rats,but not in renal tubules.Conclusions Bone marrow mesenchymal stem cells can promote the recovery of acute renal tubular epithelial cells damage caused by mercuric chloride.The mechanism may partly depend on regulating the excretion of cytokines in renal microenvironment rather than completely depend on their differentiation to tubular cells.