Objective To establish a rapid assay for genotyping of IL-6-597G/A and-572G/C polymorphisms by duplex real-time PCR assay.Methods One pair of primers and two pairs of fluorescent hybridization probes had been used to genotype two mutation sites in the promoter region of IL-6 gene by fluorescent resonance energy transfer and melting curves.Results Duplex real-time PCR method for genotyping of two mutation sites simultaneously had been developed and 123 health people samples were analyzed by this new method.The results showed that three genotypes were found in IL-6 gene-572G/C polymorphisms.They were GG,GC and CC genotypes.IL-6 gene promoter-597G/A polymorphism analysis showed 4 cases displayed GA genotype and other possessed GG genotype.No AA genotype had been found.Conclusions Duplex real-time PCR method is simple and fast.It provided an accurate and economic method which is suitable for clinical gene genotyping.

Objective The significance of serum cytokine profile in coronary heart disease patients have been evaluated by observing the variation cytokines Methods The serum IL-1 a,IL-1 ?,IL-2,IL-4, IL-6 ,IL-8,IL-10,VEGF,IFN-?,,TNF-?,MCP-1 and EGF in 76 patients and 26 matched healthy volunteers have been observed.Serum cytokines had been measured by protein chips methods in EVIDENCE 180 automatic biochips analyzer and cytokine profile were got by bioinformatics.Results The serum level of IL-1?,IL-6,IL-8,and TNF-? were (4.32?8.14) ng/L ,(13.16?16.81) rig/L,(375.53?292.14) ng/L,(15.46?15.38) ng/L respectively,which increased obviously in patients (P

0.05),but enhanced the expression of CD80,CD86 significantly(P

Objective To learn the prevalence and trends of Brucella disease in Ningxia, in order to provide scientific basis for effective control of the disease. Methods Data of Brucella cases reported through city network from 2004 to 2009 in Yinchuan city, Shizuishan city, Wuzhong city, Guyuan city and Zhongwei city of Ningxia were reviewed and retrospectively analyzed. The data included demographic characteristics, treatment conditions and medical history so on related information. Analytical indicators include reported incidence;with patients' gender, age, regional distribution, urban and rural distribution;become chronic and associated factors;distribution of the cases reporting unit and so on. Results From 2004 to 2009, Ningxia had reported 349 cases of Brucellosis, no deaths, the annual incidence rates reported were 0.017/10 million, 0.543/10 million, 0.151/101 (295/54);The proportion of 34- to 40-year-old age group was higher than other age groups(27.5%, 96/349);Occupational distribution of patients was mainly farmers and herdsmen(70.2% ,245/349), in regional distribution of the patients, the highest percentage was Wuzhong city(61.9%,216/349), followed by Yinchuan city(22.9%,80/349);The proportion of patients in rural areas(97.4% ,340/349) was higher than urban(2.6% ,9/349);the proportion of patients converted to chronic was 11.2% (39/349). With age, the chance of patients converted to chronic was in a decreasing trend(odds ratio was 0.966);cases reported by Centre for Disease Control and Prevention accounted for 74.8%(261/349), by hospital accounted for 25.2%(88/349). Conclusions The reported incidence of Brucellosis in Ningxia is in a rapid upward trend year by year, the patients is mainly young men, the rate of converted to chronic is higher and the ability of hospital in founding and reporting of the cases is weaker.Comprehensive measures should be taken to increase funding, strengthen monitoring, and continuously improve the level of awareness and diagnosis of medical personnel for further strengthen the prevention and control of Brucellosis.

OBJECTIVETo explore action mechanism of electroacupuncture (EA) at "Fenglong" (ST 40) for treatment of hyperlipidemia.

METHODSForty SPF-grade SD rats were randomly divided into a normal group (group A), a model group (group B), a model diet-control group (group C), a model + EA group (group D) and model diet-control + EA group (group E), 8 cases in each one. The rats in group A were fed with normal diet continuously while those in the rest 4 groups were fed with high-fat diet to establish hyperlipidemia model. Afterwards, the rats in group C and group E were fed with normal diet, while EA at "Fenglong" (ST 40) was applied in group D and group E, 30 min per time, once a day. After 28 days of treatment, macrophage was collected in each group. Oil red O-staining was applied to detect the formation of foam cells, and enzyme-linked immunosorbent assay (ELISE) was adopted to measure the contents of total cholesterol (TC) in macrophage and analyze the rate of cholesterol efflux.

RESULTSThe counts of positive cells of oil red O-staining and the contents of TC in the group B, group C and group D were significantly increased compared with those in the group A (all P < 0.01). The counts of positive cells and contents of TC in the group C, group D and group E were significantly clecreased compared with those in the group B (all P < 0.01), and the decline in group D was more obvious than that in the group C (all P < 0.01). Compared with group C and group D, the counts of positive cells and contents of TC in the group E was obviously decreased (all P < 0.01), which was not statistically different from group A (P > 0.05). In the meantime, compared with group C, the rate of cholesterol efflux in group D and group E was significantly increased (both P < 0.01), and the rise in group E was more obvious than that in the group D.

CONCLUSIONThe electroacupuncture at "Fenglong" (ST 40) could significantly prohibit the transformation of macrophage into foam cell and increase rate of cholesterol efflux in macrophage, which could prevent and reverse the formation of foam cell and play an essential role in treating hyperlipidemia and stopping it from developing into a further level.

Acupuncture Points ; Animals ; Biological Transport ; Cells, Cultured ; Cholesterol ; metabolism ; Electroacupuncture ; Foam Cells ; metabolism ; Humans ; Hyperlipidemias ; metabolism ; therapy ; Macrophages ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

Objective Alendronate is widely used as an anti-osteoporosis agent , while simvastatin , as a lipid-lowering drug , is being considered beneficial for bone formation .The present study aims to compare the effects of simvastatin and alendronate on bone loss in ovariectomized ( OVX) rats. Methods Thirty-two female SD rats were equally randomized into a sham operation , an OVX mod-el, a simvastatin ( OVX +S), and an alendronate ( OVX +A) group.All the rats but those in the sham operation group underwent dual ovariectomy .The animals of the OVX model and OVX +S groups were treated intragastrically with vehicle and simvastatin at 5 mg per kg of the body weight per day , respectively , while those of the OVX+A group injected subcutaneously with alendronate at 70μg per kg of the body weight per week .After 12 weeks of intervention , all the rats were sacrificed for detection of the serum concentrations of PINP and ICTP by ELISA, analysis of bone mineral density and bone histo-morphometry parameters of L 4 vertebrae , determination of the maximum loading and elastic modulus of L 5 vertebrae by compression test. Results The serum concentrations of P1NP and ICTP were significantly lower in the sham operation than in the other three groups (P<0.05), and that of ICTP markedly lower in the OVX +A than in the OVX model and OVX +S groups (P<0.05).Bone mineral density was remarkably lower in the OVX model ([0.238 ±0.007] g/cm2 ) than in the sham operation ([0.276 ±0.015] g/cm2), OVX+S ([0.250 ±0.014] g/cm2) and OVX+A groups ([0.269 ±0.014] g/cm2) (P<0.05), lower in the OVX+S than in the sham operation and OVX +A groups (P<0.05), but with no statistically significant difference between the latter two (P>0.05).Bone histomorphometry showed significantly lower BV /TV in the OVX model than in the sham operation and OVX +A groups ([19.9 ±1.69]%vs [29.03 ±2.59]%and [27.05 ±1.91]%, P<0.05), but markedly higher Tb.Sp in the former than in the latter two groups ([357.33 ±26.55] μm vs [211.17 ±16.56] μm and [252.50 ±19.35] μm, P<0.05).The maximum load and elastic modulus of L5 vertebrae were significantly lower in the OVX model rats than in the other three groups (P <0.05). Conclusion Both simvastatin and alendronate can inhibit bone loss in OVX rats , with comparable effects of preventing the deteriora-tion of biomechanical properties , but the latter is evidently more effective in maintaining bone mineral density .

Objective To investigate the effects of simvastatin on the bone loss and deterioration of bone quality with different intervention programs. Methods Thirty two 3?month?old female Sprague?Dawley ( SD ) rats were randomized into 4 groups of 8 animals in each: All rats but those in the sham group ( A) received bilateral ovariectomy, and treated by vehicle (B), simvastatin (5 mg/kg/day) at first half period (C) or at latter half period (D). The rats in group C received simvastatin by a gavage after the OVX operation immediately, and stopped at 10 weeks after OVX. The rats in group D began to receive simvastatin treatment from 10 weeks after OVX and ended at 20 weeks after OVX. At week 20, all rats were sacrificed and the concentrations of serum PINP and ICTP were detected by ELISA, L3 vertabra was used to detect the bone mineral density, L4 vertebra was used to analyze the maximum loading and elastic modulus by compression test, and the microstructure of the L5 vertebra was detected by micro?computed tomography. Results 1. ELISA analysis:the concentrations of serum P1NP and ICTP in the groups A, B and C were significantly higher than that of group A (P<0?05). 2. BMD test showed that the rats in group B had significantly lower BMD than the other 3 groups (P<0?05), while the BMD of groups C and D were markedly lower than that of group A (P<0?05). 3. Biomechanical test:the maximum load and elastic modulus of L4 vertebrae in the group B were significantly lower than the other 3 groups ( P<0?05), and those of the groups C and D were markedly lower than that in the group A (P<0?05). 4. micro?CT:BV/TV and Tb. N in the sham operated rats were significantly higher than those of the other 3 groups, while the opposite trends was found in Tb. Sp (P<0?05), and the Tb. Sp in the groups C and D were significantly lower than that of group B (P<0?05). Conclusions Our data demonstrate that bone loss and deterioration of bone micro?structure and biomechanical properties occurred at 20 weeks after ovariectomy, both the first?half?period and latter?half?period treatment by simvastatin may partially prevent these changes, but can not restore the BMD to normal level.

Objective:To observe the effects of acupuncture on the characteristics of neuro-electrophysiological activity in hippocampal CA1 and CA3 areas of rats with post-traumatic stress disorder (PTSD).Methods:Fifty Sprague-Dawley (SD) rats were randomly divided into a blank group,a model group,a grasping group,a Western medicine group and an acupuncture group,with 10 rats in each group.Except for the blank group,rats in the other 4 groups all received the combined stress modeling method.Rats in the Western medicine group were intragastrically administrated with paroxetine hydrochloride,those in the acupuncture group received acupuncture intervention,those in the grasping group received grasping fixation,and those in the model group and the blank group did not receive any interventions.After 14 d of intervention,the interspike interval (ISI) and power spectral densities (PSD) were analyzed and mapped by in vivo multiple channels to record the neuron clusters discharge in the hippocampal CA1 and CA3 areas.Results:Compared with the blank group,ISI was prolonged in the CA1 and CA3 areas of the model group and the grasping group,and the concentrated PSD distribution area moved down (P＜0.05 or P＜0.01).Compared with the grasping group,the ISI of the CA1 and CA3 areas in the Western medicine group and the acupuncture group was shortened,and the concentrated PSD distribution area moved up (P＜0.05 or P＜0.01).The ISI and PSD distributions in the CA1 and CA3 areas of the acupuncture group were not statistically different from those in the Western medicine group (both P＞0.05).Conclusion:Both acupuncture and paroxetine hydrochloride can significantly regulate the neuro-electrophysiology activity of hippocampal CA1 and CA3 areas in PTSD rats,which may be one of the mechanisms of acupuncture intervention to promote PTSD recovery.

OBJECTIVETo observe the effect of Bushen Wenyang Huayu Recipe (BWHR) on hypoxia inducible factor-1α (HIF-1α), proline hydroxylase2 (PHD2), von Hippel Lindau disease (VHL) suppressor gene expressions in endometriosis (EM) rats with Shen yang deficiency blood stasis syndrome (SYDBSS), and to explore the pathogenesis of EM and the mechanism of BWHR for treating EM.

METHODSTotally 50 SD rats were randomly divided into five groups, i.e., the blank control group, the sham-operation group, the model group, the Chinese medicine (CM) group, and the Western medicine (WM) group, 10 in each group. Rats in the blank control group and the sham-operation group were fed routinely. Rats in the rest 3 groups received 30-day "extended refrigerator freezing and ice water immersion" and combined with " autotransplantation" to establish EM rat model with SYDBSS. One Milliliter BWHR at 3.33 g/mL was administered to rats in the CM group by gastrogavage. Gestrinone at the daily dose of 0. 5 mg/kg was administered to rats in the WM group by gastrogavage. Equal volume of normal saline was administered to rats in the model group, the blank control group, and the sham-operation group. The size and morphology of ectopic foci in rats were observed after 4 weeks of medication. Expressions of serum CA125, plasma cyclic adenosine monophosphate (cAMP), and plasma cyclic guanosine monophosphate (cGMP) were detected by radioimmunoassay. Morphological changes of eutopic endometrium and ectopic tissue were observed under the optical microscope by HE staining. Protein expressions and contents of HIF-lα, PHD2, and VHL were detected by immunohistochemical SABC method and Western blot. mRNA expressions of HIF-1α, PHD2, and VHL were detected by RT-PCR.

RESULTSThe ectopic foci grew significantly in the model group. Their volumes were obviously contracted after treated by CM and WM. Compared with the blank control group and the sham-operation group, serum CA125 and plasma cGMP obviously increased, cAMP obviously decreased (P < 0.05); expressions and contents of HIF-1α mRNA and protein all decreased (P < 0.05); mRNA and protein expressions and contents of PHD2 and VHL all decreased in the model group (P < 0.05). Compared with model group, levels of CA125 and cGMP obviously decreased; cAMP levels obviously increased, expressions and contents of HIF-1α mRNA and protein all increased, mRNA and protein expressions and contents of PHD2 and VHL all increased in the WM group and the CM group (P < 0.05). Compared with the CM group, PHD2 protein contents were higher in the WM group (P < 0.05). HIF-1α was negatively correlated with PHD2 (r = -0.799, P = 0.00). HIF-1α was negatively correlated with VHL (r = -0. 625, P = 0.003).

CONCLUSIONSBWHR could effectively treat EM. Its mechanism might be associated with reducing contents of HIF-1α, serum CA125, and plasma cGMP, and up-regulating expressions of PHD2, VHL, and cAMP.

Animals ; Cyclic AMP ; Drugs, Chinese Herbal ; therapeutic use ; Endometriosis ; drug therapy ; metabolism ; Female ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Proline ; metabolism ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Up-Regulation ; Yang Deficiency ; drug therapy ; metabolism

A processing method to enhance thrombolytic effect of Carthamus tinctorius using a fermentation technology with bacillus sp. C2-13 was investigated. The fibrinolysis and anticoagulation activity of thrombolytic extracts from an optimized fermentation process was studied using a carrageenan induced mice model. The fermented extracts resulted in significantly better thrombolytic activity, suggesting that the process was promising for use in the study and preparation of nature medicines.
Animals ; Bacillus ; Carthamus tinctorius ; chemistry ; microbiology ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Fermentation ; Fibrinolysis ; drug effects ; Fibrinolytic Agents ; pharmacology ; Male ; Mice ; Partial Thromboplastin Time ; Prothrombin Time ; Rats ; Rats, Sprague-Dawley ; Thrombin Time