OBJECTIVETo explore the effects of Wenyang Huazhuo Tongluo Recipe (WYHZTLR) containing serum on transforming growth factor β1 (TGF-β1)/Smad signaling pathway of skin fibroblasts in systemic sclerosis (SSc).

METHODSTotally 36 SSc patients were randomly assigned to Chinese medicine (CM) group, Western medicine (WM) group, and integrative medicine (IM) group according to random digit table, 12 in each group. Patients in the CM group took WYHZTLR decoction (one dose per day). Patients in the WM group took penicillamine tablet (0. 125 g each time, bid) and Prednisone Acetate Tablet (PAT 20 mg, qd). Patients in the IM group took penicillamine, PAT, and WYHZTLR decoction (in the same dosage of corresponding drugs as aforesaid). All patients were treated for one month to get drug containing serum. Besides, 10 untreated SSc patients' serum was taken as the control group. Healthy subjects' skin fibroblasts were originated from healthy skin tissue of the upper arms of 2 female patients undergoing plastic surgery. Corresponding serum of each group was added in the culture system of SSc patients' and healthy subjects' dermal fibroblasts respectively. Expression levels of TGF-β1 receptor type I (TGF-β1 RI), TGF-β1 receptor II (TGF-β1 RII), p-Smad2/3 and Smad7 protein were examined by Western blot. Expression levels of collagen type I and collagen type III (Col-I, Col-III) mRNA were examined by reverse transcription PCR. Contents of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinases-1 (TIMP-1) in the supernatant of SSc, skin fibroblasts were examined by ELISA.

RESULTSCompared with the control group, expression levels of TGF-β1 R I and p-Smad2/3 protein significantly decreased, but expression levels of Smad7 protein significantly increased in skin fibroblasts of SSc patients and healthy subjects of WM, CM, and IM groups (P <0.05, P <0. 01). Meanwhile, the expression level of TGF-β1 RII decreased in the IM group (P <0. 05, P <0. 01). Compared with the WM group, expression levels of TGF-β1 RI and p-Smad2/ 3 protein significantly decreased, but that of Srnad7 protein significantly increased in IM groups (P <0. 01). mRNA expression levels of Col-I and Col-II in SSc skin fibroblasts significantly decreased more in WM, CM, and IM groups than in the control group (P <0. 05, P <0. 01). Besides, the expression level of Col-III mRNA was significantly lower in the IM group than in the WM group (P <0.01). Compared with the control group, serum levels of MMP-9 and MMP-9/TIMP-1 ratios increased more obviously in WM, CM, and IM groups (P <0.05, P <0.01). But expression levels of TIMP-1 decreased significantly in CM and IM groups (P <0.01). Expression levels of MMP-9 and MMP-9/TIMP-1 ratios increased more in the IM group than in the WM group (P <0. 01). Expression levels of TIMP-1 decreased more in CM and IM groups than in the WM group (P <0.01).

CONCLUSIONWYHZTLR containing serum could reduce expression levels of Col-I and Col-III possibly through regulating key signal molecules, such as TGF-β1 RI, p-Smad2/3, and Smad7 in TGF-β1/Smad signaling pathway of SSc skin fibroblasts, and inhibiting transduction of TGF-β1/Smad signaling pathway.

Collagen Type I ; Collagen Type III ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Fibroblasts ; Humans ; Matrix Metalloproteinase 9 ; Scleroderma, Systemic ; drug therapy ; metabolism ; Signal Transduction ; drug effects ; Smad2 Protein ; Smad7 Protein ; Transforming Growth Factor beta1 ; metabolism

Objective To investigate the distribution of water-borne fluoride and the current status of water defluoridation project by improving drinking water quality in endemic fluorosis areas in Shandong province,and to provide scientific basis for making strategies in prevention and control of the disease. Methods According to "the National Technical Scheme for Endemic Disease Control in 2004, 2005 and 2006", 113 endemic fluorosis diseased counties(cities, and districts) of the province's 17 cities were screened in order to investigate the fluoride level in drinking water in fluorosis villages, recheck the fluoride level after implementing the water defluoridation project, and investigate the current status of the water defluoridation project. The fluoride level in drinking water was determined by F-ion selective electrode. Results There were a total of 5816 water defluoridation projects in the province. Most of them were carried out by drilling a deep well to get under-ground water. The wells still in good condition were accounted for 72.80% (4234/5816). Intermittent operated wells were accounted for 3.11% (181/5816). Abandoned wells were accounted for 24.09%(1401/5816). Level of water fluoride was determined in 6940samples from fluorosis villages(villages that not carry out the water defluoridation project as well as villages carried out the water defluoridation project with abandoned wells were included) and the value that lower or equal to 1.00mg/L was determined in 2987 villages which accounted for 43.04% (2987/6940). Level of water fluoride that over 1.00 mg/L was found in 3953 villages which accounted for 56.96% (3593/6940), and the highest level of water fluoride was 11.33 mg/L. Level of water fluoride were determined in 4415 samples from water defluoridation project and the value lower or equal to 1.00 mg/L was in 2983 wells which accounted for 65.53%(2983/4415). The value over 1.00 mg/L was in 1522 wells which accounted for 34.47%(1522/4415), the highest value of water fluoride was 9.71 mg/L. Conclusions Level of water fluoride in up to 1/2 of the villages and 1/3 of the projects, is still higher than the standard in Shandong province. Nearly 1/4 of the project has been abandoned. The current situation for endemic fluorosis control is still not good in the province, countermeasures for endemic fluorosis must be carried out as soon as possible and surveillance of water defluoridation project must be strengthened.

OBJECTIVETo observe the effect of Wenyang Huazhuo Tongluo Recipe (WYHZTLR) on the proportion of T helper 17 cells (Thl7)/regulatory T cells (Treg), and serum levels of IL-17 and IL-10 in peripheral blood of systemic sclerosis (SSc) patients with yang qi insufficiency and turbidity induced collaterals blockage syndrome (YQITICBS).

METHODSTotally 82 SSc patients were randomly assigned to the Western medicine group (as the control group) and the integrated Chinese and Western medicine group (as the treatment group), 41 cases in each group. All patients took methotrexate (MTX) tablet and prednisone tablet. Patients in the treatment group additionally took WYHZTLR. The treatment course for all was six consecutive months. Besides, another 70 healthy volunteers were recruited as a healthy control group (as the healthy group). Percentages of Th17 and Treg in peripheral blood were detected by flow cytometry. Serum levels of IL-17, IL-10, von Willebrand factor (vWF), aminoterminal propeptide of type l procollagen (PIIINP), and cross-linked carboxyterminal telopeptide of type I collagen ( I CTP) were measured by double antibody sandwich ELISA. The correlations between Th17/Treg and levels of vWF, PIIINP, I CTP, skin score, and disease activity index were observed by Pearson correlation analysis.

RESULTSThe percentage of Th17 in peripheral blood, ratios of Th17/Treg, and the serum level of IL-17 were significantly higher, but the percentage of Treg and the serum level of IL-10 were significantly lower in SSc patients, when compared with those of the healthy group (P <0. 05, P <0. 01). Compared with the same group before treatment, the percentage of Thl7, ratios of Thl7/Treg, and levels of IL-17, vWF, and PIIINP all decreased in the two groups after treatment (P <0.05, P <0.01), but the percentage of Treg and the IL-10 level increased (P <0. 05, P <0. 01). Meanwhile,the level of I CTP was higher in the treatment group after treatment (P <0. 05). The improvement of all indices except the percentage of Th17 was more obvious in the treatment group than in the control group (P <0. 05, P <0. 01). The ratio of Th17/Treg was positively correlated with levels of vWF, PIIINP, skin score, and disease activity index before and after treatment respectively (P <0. 01), but with no obvious correlation with the level of I CTP (P >0. 05).

CONCLUSIONWYHZTLR could achieve its therapeutic effect on SSc patients by regulating Th17/Treg imbalance, lowering levels of vWF and PIIINP, and elevating the level of I CTP.

Drugs, Chinese Herbal ; therapeutic use ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Humans ; Interleukin-10 ; Interleukin-17 ; Methotrexate ; Peptide Fragments ; Procollagen ; Scleroderma, Systemic ; drug therapy ; T-Lymphocytes, Regulatory ; Th17 Cells

BACKGROUND:Sepsis-induced myocardial injury is one of the major predictors of morbidity and mortality of sepsis. The cytoprotective function of erythropoietin (EPO) has been discovered and extensively studied. However, the cardioprotective effects of EPO on sepsis-induced myocardial injury in the rat sepsis model has not been reported.METHODS:The rat models of sepsis were produced by cecal ligation and perforation (CLP) surgery. Rats were randomly (random number) assigned to one of three groups (n=8 for each group):sham group, CLP group and EPO group (1000 IU/kg erythropoietin). Arterial blood was withdrawn at 3, 6, 12, and 24 hours after CLP. cTnI, BNP, CK-MB, LDH, AST, TNF-α, IL-6, IL-10, and CRP were tested by the ELISA assay. Changes of hemodynamic parameters were recorded at 3, 6, 12, 24 hours after the surgery. Histological diagnosis was made by hematoxylin and eosin. Flow cytometry was performed to examine cell apoptosis, myocardium mitochondrial inner membrane potential, and NF-κB (p65). Survival rate at 7 days after CLP was recorded.RESULTS:In the CLP group, myocardial enzyme index and inflammatory index increased at 3, 6, 12 and 24 hours after CLP compared with the sham group, and EPO significantly blocked the increase. Compared with the CLP group, EPO significantly improved LVSP, LV +dp/dtmax, LV -dp/dtmin, and decreased LVEDP at different time. EPO blocked the reduction of mitochondrial transmembrane potential, suppressed the cardiomyocyte apoptosis, inhibited the activation of NF-κB, and reduced the production of proinflmmatory cytokines. No difference in the survival rate at 7 days was observed between the CLP group and the EPO group.CONCLUSION:Exogenous EPO has cardioprotective effects on sepsis-induced myocardial injury.

OBJECTIVETo detect the immunoreactivity of targeted fusion anti-caries DNA vaccine pGJA-P in vitro, and the ability to enhance the immune responses compared with the non-targeted fusion anti-caries DNA vaccine pGLUA-P.

METHODSThe CHO cells were transfected with pGJA-P and the expression of recombinant protein in cultured supernatants were detected using Western blotting. 5 to 6-month-old female Japanese rabbits were immunized with either pGJA-P or pGLUA-P via either intramuscular injection (i.m.) or intranasal route (i.n.). The sera and saliva were collected and the antibody responses were checked by ELISA. The effect of immune sera on the synthesis of water-insoluble glucan by glucosyltransferase of S. mutans was examined.

RESULTSThe expressed protein could response to specific anti-GTF antibody. The antibody responses in serum generated by pGJA-P via i.m. were significantly higher than those generated by pGLUA-P (P < 0.01). The antibody responses in saliva generated by pGJA-P via i.n. were significantly higher than those generated by pGLUA-P (P < 0.01). The higher mucosal antibody response induced by pGJA-P via i.m. compared with pGLUA-P (P < 0.01) was detected. The immune sera of rabbits immunized by pGJA-P via i.m. most significantly inhibited the synthesis of water-insoluble glucan by glucosyltransferase.

CONCLUSIONSThe recombinant protein expressed by pGJA-P had the immunoreactivity to anti-GTF antibody. pGJA-P could induce faster and higher specific mucosal SIgA antibody responses via i.n. or serum IgG antibody responses via i.m. compared with non-targeted DNA vaccine, pGLUA-P. High titres of specific mucosal antibodies were found in rabbits immunized with pGJA-P via i.m. The immune sera of rabbits immunized by pGJA-P via i.m. displayed the ability of inhibiting the synthesis of water-insoluble glucan by glucosyltransferase.

Animals ; Bacterial Proteins ; immunology ; CHO Cells ; Cricetinae ; Dental Caries ; prevention & control ; Female ; Glucosyltransferases ; immunology ; metabolism ; Immunoglobulin G ; blood ; Membrane Glycoproteins ; immunology ; Rabbits ; Recombinant Fusion Proteins ; administration & dosage ; immunology ; Streptococcus mutans ; immunology ; Transfection ; Vaccines, DNA ; administration & dosage ; immunology

OBJECTIVETo investigate the level of occupational exposure to 5-fluorouracil (5-Fu) in the pharmacy intravenous admixture service (PIVAS) of a hospital, and identify the sources of 5-Fu contamination.

METHODSThe 5-Fu concentrations in air, on the surface of different areas in PIVAS and personal protective equipments were detected using UV-vis spectrophotometry.

RESULTSThe 5-Fu in air could not be detected. The 5-Fu concentrations on five different surfaces of biological safety cabinets were (22.00 +/- 6.35), (13.99 +/- 2.46), (14.13 +/- 0.72), (7.25 +/- 1.19) and (9.87 +/- 1.23) ng/cm2, respectively, which were significantly higher than those [(3.14 +/- 0.04), (5.43 +/- 0.65), (2.26 +/- 0.17), (2.26 +/- 0.17) and (3.63 +/- 0.46) ng/cm2] of corresponding controls (P < 0.05 or P < 0.01). The 5-Fu concentrations of the floor under cabinets [(18.19 +/- 5.22) ng/cm2], the floor in front of cabinets [(10.25 +/- 2.57)ng/cm2], the office floor [(11.64 +/- 2.53) ng/cm2], the terrace floor [(99.89 +/- 14.06 ) ng/cm2], the floor beside trash can in dressing room [(24.54 +/- 0.23) ng/cm2] were significantly higher than those of control [(3.36 +/- 0.11 ) ng/cm2] (P < 0.05 or P < 0.01). The 5-Fu concentrations of the tables in preparation room [(7.22 +/- l.04) ng/cm2] and the tables in office [(11.81 +/- 1.18) ng/cm2] were significantly higher than those of control [(5.56 +/- 0.14) ng/cm2] (P < 0.05 or P < 0.01). The 5-Fu concentrations of the indoor handle in preparation room were significantly higher than those of controls (P < 0.05 or P < 0.01). 5-Fu concentrations on the surfaces of outdoor handle and floor beside door in preparation room were not significantly increased compared with controls (P > 0.05). The 5-Fu concentrations on the surfaces of infusion bags, transfer box, transfer trays were significantly higher than those of controls (P < 0.05). The differences of 5-Fu concentrations between outer and inner masks and controls were not significant (P > 0.05). The 5-Fu concentrations of gloves of preparing and checking staffs were significantly higher than those of controls (P < 0.05 or P < 0.01).

CONCLUSIONThe preparing and checking process of 5-Fu and the treatment of medical wastes are major sources of 5-Fu contamination.

Antineoplastic Agents ; analysis ; Drug Administration Routes ; Fluorouracil ; analysis ; Humans ; Occupational Exposure ; Pharmacy Service, Hospital

Objective To investigate the current status of endemic fluorosis in Shandong province, and to provide the scientific evidence for making strategies for prevention and control of the disease. Methods According to "The National Technical Scheme for Endemic Disease Control in 2008", thirty-four counties were divided into mild, moderate and severe endemic fluorosis areas and a village was randomly selected from each category of the area to carry out the monitoring of endemic fluorosis. The content of fluoride in drinking water and urine was determined by F-ion selective electrode, dental fluorosis of children aged 8 to 12 was diagnosed by Dean method and skeletal fluorosis diagnosed by clinic and X-rays. Results The monitoring was done in 70 water-improving villages in 34 counties, among which 54 villages had water fluoride content ≤ 1.00 mg/L and accounted for 77.14%(54/70), 16 villages had water fluoride content ＞ 1.00 mg/L and accounted for 22.86%(16/70), the highest water fluoride content was 4.46 mg/L. The monitoring was also carried out in 32 non-water-improving villages in 34 counties, among which 9 villages had water fluoride content ≤ 1.00 mg/L and accounted for 28.12%(9/32), 23 villages had water fluoride content ＞ 1.00 mg/L and accounted for 71.88% (23/32), the highest water fluoride content was 4.09 mg/L. The total rate of dental fluorosis of children aged 8 to 12 was 45.81%(1988/4340), the index of dental fluorosis was 0.97 and the rate of dental damage was 6.91%(300/4340). The urinary fluoride values above 1.40 mg/L were found in 55.33%(1417/2657) of children aged 8 to 12, with the highest urinary fluoride concentrations was 18.53 mg/L. The rate of skeletal fluorosis by clinic and X-rays in adults older than 16 years were 4.25% (2462/57 968) and 28.40%(23/81 ), respectively. The urinary fluoride values above 1.60 mg/L were found in 55.86% (1130/2023) of adults older than 16 years, with the highest urinary fluoride concentrations was 25.44 mg/L. Conclusions Endemic fluorosis in Shandong province has not yet been effectively controlled,control situation is still grim. Prevention efforts need to be further strengthened.

Objective To investigate the present situation of water-improving defluoridation project and the distribution of water fluoride in fluorosis areas in Shandong province. Methods In 2005 - 2007, according to "The National and Shandong province Technical Scheme for Endemic Disease Control", the water-improving form,water type and the running status of defluoridation project in 17 cities of Shandong province were investigated, and the water-fluoride of the running projects were determined by F- ion selective electrode. Results A total of 5816projects were built that involved 8776 villages, 110 counties, 17 cities distributed in Shandong province. In which,the drilling of water-improving projects accounting for 94.55% (5499/5816), under-ground water type 97.73%(5684/5816);operating normally accounting for 75.91% (4415/5816), 7246 villages were supplied with the water, beneficiary population 6 946 459 people, non-normal projects accounting for 24.09%(1401/5816), including 1530 diseased villages. There were still 4415 projects running well. The projects with water fluoride lower than 1.0mg/L were 2893, accounting for 65.53% (2893/4415). The water fluoride higher than 1.0 mg/L accounted for 34.47%(1522/4415), ＞ 2.0 mg/L of 548, ＞ 4.0 mg/L of 97, and the maximum value of water fluoride was 9.71mg/L. Conclusions Nearly 1/4 of the water-improving projects in Shandong province are abnormal. The water fluoride of the project of more than 1/3 are exceeding the standard( ＞ 1.0 mg/L) in 4415 projects, and water-fluoride of 97 projects are higher than 4.0 mg/L. More measures should be taken to strengthen the supervision of the projects and monitoring of water fluoride.

Objective To understand the current status of the prevention and control of endemic fluorosis in the southwestern area of Shandong province. Methods In 2007, the progress of water-improving defluoridation, the operating state and water fluoride content of the water-improving project, which was determined by fluorosis selective ion electrode, and the inhabitant related indexes of endemic fluorosis were extensively surveyed in the three main fluorosis counties-Jiaxiang, Yuncbeng and Liangshan of the southwestern area of Shandong province. Results Among 1371 fluorosis villages in the 3 counties, 53.61%(735/1371) of which had undergone water-improving defluoridation, the rate in Jiaxiang, Yuncheng and Liangshan being 38.0% (220/579),65.51% (378/577) and 63.72% ( 137/215 ) respectively; the normally functioning rate of this project was 76.73% (564/735), projects out of order accounted for 23.27% (171/735). Among 263 well-functioning projects from the three counties, the rate with water fluoride higher than 1.0 mg/L was 35.36%(93/263), the maximum value being 4.17 mg/L. The urine fluoride content of 440 children aged 8 - 12 years and 484 adults over 30 years old were examined in 13 fluorosis villages of the three counties, the geometric mean was 2.98,3.06 mg/L respectively; the individual maximum was 12.83,14.49 mg/L respectively; the detectable rate of dental fluorosis among children aged 8 - 12 was 84.28% (649/770) ,17.66%(136/770) had defect and the index of dental fluorosis was 1.89; the rates of the clinical and X-ray skeletal fluorosis of the adults aged more than 30 were 44.40%(234/527) and 24.67%(130/527) respectively, and the abnormal electrocardiography rate was 32.43% (168/518) in the adult, mostly T-wave abnormality. Conclusions The progress of the water-improving defluoridation in the southwestern area of Shandong province was relatively slow, the water fluoride content of the water-improving projects seriously exceeded standard, and the condition of the fluorosis had not been effectively controlled.

Objective To understand the fulfillment of control measures of endemic fluorosis and find out the prevalent trend of this disease in Shandong Province from 1992 to 2006 in order to provide a scientific basis for the set up of control strategies. Methods According to "the National Survey Scheme of Endemic fluorosis", Huantai County in 1992-1996 and Liaagshan County in 1997-2006 were selected, where water-improving defluoridation and the contents of water fluoride were searched in two counties; dental fluorosis and urine fluoride in children aged 8-12 years old, clinical and X-ray skeletal fluorosis in adult over 16 years old were searched respectively in Lijia Village of Huantai County and Dongxu Village of Liangshan County. Results One hundred percent(304/304) high-fluoride villages of Huantai County in 1992 and 63.72% (137/215) villages of Liangshan County in 2006 had finished the project of water-improving defluoridation; the rate of high-fluorine water(>1.0 mg/L) in normal operation projects was 5.00%(5/100)-17.14%(18/105) in Huantai County from 1992 to 1996, and 18.97%(11/58)-45.61%(26/57) in Liangshan County from 1997 to 2006. The water fluoride of Lijia Village was<1.0 mg/L from 1992 to 1996, the water fluoride of Dongxu Village was<0.5 mg/L from 1997 to 2006. The detection rate of dental fluorosis of children aged 8-12 years at Lijia Village dropped from 30.86%(25/81) in 1992 to 10.13%(8/79) in 1996(χ2=12.41, P<0.05), and the Dongxu Village dropped from 68.95%(151/219) in 1997 to 0(0/38) in 2006(χ2=222.04, P<0.01). In the Lijia and Dongxu Villages, urine fluoride in children aged 8-12 years old was lower than 1.40 mg/L. In the adults over 16 years of the two villages, the positive rate of X-ray skeletal fluorosis of Lijia Village dropped from 30.0%(15/50) in 1992 to 13.95%(6/43) in 1996(χ2=3.41, P>0.05), and all the cases were in degree I, the rate of Dongxu Village dropped from 64.58%(31/48) in 1997 to 16.67% (4/24) in 2006(χ2=14.71, P<0.01), and no eases of degrees Ⅲ was detected. Conclusions The progress of water defluoridation was uneven in 2 counties, slow in Liangshan County; and the water fluoride in part of the project exceeded standard; the water fluorid of 2 surveyed villages was normal and the endemic fluorosis was under control. More financial input is requested to accelerate the process of water defluoridation and to reinforce the management of the projects and illness monitoring, thus to reduce the harm of fluorosis to the limit.