OBJECTIVE:To establish the method for the determination of plasma concentration of magnesium isoglycyrrhiz-inate in portal vein and peripheral venous blood of patients underwent liver resection,to further validate and evaluate pharmacoki-netic characteristics,rational and safe use of drugs in the clinic. METHODS:31 patients underwent liver resection in our hospital during Oct. 2014-Mar. 2015 were given magnesium isoglycyrrhizinate intravenously at the beginning of surgery. Portal vein and pe-ripheral venous blood of patients were drawn at 1 hour after drug use,and HPLC-UV detection method was used to determine the plasma concentration of drug. RESULTS:The retention time of isoglycyrrhizinate magnesium was 4.5 min,which showed a good peak shape,and was not interfered with the determination by plasma endogenous peak. The plasma concentration ranged from 0.55 to 55.00 mg/L. The minimum quantitative concentration was 0.55 mg/L. The extraction recoveries were 84.7%-87.1%,and method recoveries were 101.2%-105.4%,and RSDs of intra-day and inter-day were less than 6%. Plasma concentration of magnesium iso-glycyrrhizinate in portal vein blood was significantly higher than in peripheral vein blood of patients underwent liver resection (close to 2 times);and plasma concentration was not affected by primary liver diseases and underlying diseases such as cirrhosis. CONCLUSIONS:The method is simple and has high recovery rate of extraction,high accuracy and high sensitivity. It can meet the needs of pharmacokinetic study. After the application of magnesium isoglycyrrhizinate during liver resection,there is higher blood concentration of magnesium isoglycyrrhizinate in portal vein,which is beneficial to protect liver cells and improve liver func-tion. It is suitable during perioperative period of liver.

Objective To investigate the expression of CD133 protein in primary lesions of hepatocellular carcinoma (HCC) and its clinical significance.Methods The expression of CD133 protein in 190 patients with HCC was detected by immunohistochemical staining ABC method.The correlation of CD133 protein expression with the clinicopathologic parameters and features after operation was analyzed.Results The expression positive rate of CD133 protein in cancer tissues was 22.1％ (42/190).There was significant correlation between the expression of CD133 protein and tumor differentiation (P ＜ 0.001),micro vessel invasion (P =0.016) and hepatitis B virus infection (P =0.024).Univariate analysis of factors demonstrated that differentiation level,lymph node,macro vessel invasion,micro vessel invasion,TNM stage and CD133 expression were correlated with disease-free survival after surgery (all P ＜ 0.05).Multivariate analysis of factors demonstrated that both CD133 expression and micro vessel invasion were independent prognostic factors of disease-free survival after surgery.Prognostic analysis demonstrated that the disease-free survival of CD133 positive group was significantly lower than that of CD133 negative group.Conclusion The CD133 protein expression in HCC tissues is related with development,metastasis and prognosis of HCC.

Objective To investigate the ultrasonic appearance of peripheral neurilemmoma.Methods Ultrasonic appearances of 43 patients with 46 neurilemmomas confirmed by surgery and pathology were analyzed retrospectively and compared with their MRI and pathological features.Results Forty-five neurilemmomas were with regular shape and clear boundary,except one at the root of nerve C4 that was irregular in shape and unclear in boundary,like adumbbell.Sonography showed that there were 30 neurilemmomas displaying as solid tumors,15 as cystic and solid,and 1 as entirely cystic.Posterior echoenhancement,target sign,rat tail sign and vessels accompanying sign were found in 39,21,23 and 17 neurilemmomas,respectively.There were 4 neurilemmomas with ultrasound blood flow grade 0,9 with grade Ⅰ,22 with gradeⅡ,and 11 with grade Ⅲ.The target signs were found in 21 cases (21/46,45.6％) and 26 cases (26/46,56.5 ％) by ultrasound and MRI,respectively.The capability of ultrasound finding the target sign in neurilemmoma was comparable to MRI(Kappa =0.527,P ＜0.001).The diagnostic accuracy rate of neurilemmoma by ultrasonography was 73.9％(34/46).Conclusions The ultrasound has a relatively high detection of the target sign in neurilemmoma,which may provide a new basis for the ultrasonic diagnosis of neurilemmoma.

BACKGROUND:Tougu Xiaotong Capsule (TGXTC) is a clinical prescription for the treatment of osteoarthritis;however, its mechanism has not been ful y elucidated. Urokinase-type plasminogen activator (uPA) system participating in the degradation of the extracel ular matrix of articular cartilage and hyperplasia of joint synovium plays an important role in the pathological process of osteoarthritis. OBJECTIVE:To investigate the effects of TGXTC medicated serum on the expression of uPA, uPA receptor (uPAR), plasminogen activator inhibitors (PAIs), matrix metal oproteinase-3 (MMP-3), interleukin-1 beta (IL-1β) and tumor necrosis factor-α(TNF-α) in osteoarthritis synovial cel s of rats and to discuss the mechanism by TGXTC medicated serum prevents and cures osteoarthritis. METHODS:Rat models with knee osteoarthritis were established by injecting 4%papain into the knee joint cavity. Primary synoviocytes and osteoarthritis synoviocytes were cultured with col agenase digestion method. The cultured synoviocytes were divided into normal group, model group and TGXTC group. The western blot method was adopted to detect uPA, uPAR, PAI, MMP-3, IL-1βand TNF-αprotein expression of synoviocytes after acting by TGXTC medicated serum for 72 hours. RESULTS AND CONCLUSION:The expression of uPA, uPAR, MMP-3, IL-1βand TNF-αwere decreased, while PAI was increased in the TGXTC group, and there were significant differences when compared with model group. In a word, TGXTC can significantly inhibit the expression of uPA, uPAR, MMP-3, IL-1β, TNF-α, and improve PAI expression in synoviocytes, which may partly explain the mechanism of the treatment of Tougu Xiaotong Capsule on osteoarthritis.

We analyzed the epidemiological investigation results of the first case of human infected with avian influenza (H5N6) ,and summarized epidemiological characteristics of this case to provide reference to H 5N6 case screening and investiga‐tion in future works .We retrospectively described and analyzed the investigation progress ,specimen detection results of epi‐demic spot and intimate contact people ,and summarize epidemiological characteristics of this case .Result showed that the first case of human infected with H5N6 in Yunnan was a single case ,and did not showed human‐to‐human transmission .This case had contacts with wild bird and had birds’ market activity before diagnoses ;some environment specimen of birds market near patient’s home has detection results of H5N6 positive .This is a cases of local infection and did not show human‐to‐human transmission .From the investigation ,we could deduced that the transmission route is from birds to human or from birds to en‐vironment and then to human .

BACKGROUND:Tougu Xiaotong Capsule has pretty good clinical therapeutic effect on osteoarthritis of early and middle periods. However, the mechanism of Tougu Xiaotong Capsule is not ful y clarified. The RhoA GTPases can regulate chondrocyte apoptosis and hypertrophy.
OBJECTIVE:To observe the Tougu Xiaotong Capsule on the expression of Rac1and Cdc42 in tumor necrosis factor-α-induced in vitro cultured rat articular chondrocytes, and to explore its mechanism of action for combating osteoarthritis.
METHODS:Knee cartilage of the 4-week-old Sprague-Dawley rats was used to stably establish in vitro culture system of chondrocytes. Passage 3 chondrocytes were identified by toluidine blue staining. Chondrocyte apoptosis was successful y induced by 20μg/L tumor necrosis factor-αand then Tougu Xiaotong Capsule at different dosage (500, 100, 20 mg/L) was given after 24-hour incubation. MTT assay was used to detect cellsurvival, flow cytrometry to measure mitochondrial membrane potential, and western blot assay to determine the protein expression of Rac1, Cdc42, Bax and Bcl-2.
RESULTS AND CONCLUSION:Tougu Xiaotong Capsule could reduce tumor necrosis factor-α-induced apoptosis of chondrocytes to improve the survival rate of the cells, and at the same time, could down-regulate the protein expression of Rac1, Cdc42 and Bax and increase the protein expression of Bcl-2 significantly (P<0.05). Tougu Xiaotong Capsule possibly plays a therapeutic efficacy on osteoarthritis by reducing promote apoptosis Rac1, Cdc42 and Bax expression and increasing apoptosis inhibiting gene Bcl-2 expression, thereby to inhibit apoptosis of chondrocytes.

BACKGROUND:Tougu Xiaotong capsule is the clinical prescription for the treatment of osteoarthritis, however, its mechanism has not been fuly elucidated. Urokinase type plasminogen activator system which participated in the degradation of the extracelular matrix of articular cartilage and hyperplasia of joint synovium plays an important role in the pathological process of osteoarthritis. OBJECTIVE: To determine the effect ofTougu Xiaotong capsule on urokinase-type plasminogen activator system in knee cartilage tissues of knee osteoarthritis rats. METHODS: Of 144 Sprague-Dawley rats, 120 rats were randomly made into models of knee osteoarthritisvia intra-articular injection of papain, and randomly assigned to model group,Zhuanggu Guanjie Wan group [1.2 g/(kg?d)], low-doseTougu Xiaotong capsule group [0.092 g/(kg?d)], moderate-doseTougu Xiaotong capsule group [0.184 g/(kg?d)] and high-doseTougu Xiaotong capsule group [0.368 g/(kg?d)]. Each group contained 24 rats. Every 2 weeks was considered as a course, with a 2-day interval, totaly 4 courses. The remaining 24 normal rats were included in the blank group. After every two courses, a batch of experimental animals was sacrificed. The pathological changes were observed folowing staining with hematoxylin and eosin. The positive cels of urokinase-type plasminogen activator, urokinase-type plasminogen activator receptor and plasminogen activator inhibitor were measured by immunohistochemistry. The protein levels of urokinase-type plasminogen activator, urokinase-type plasminogen activator receptor and plasminogen activator inhibitor were measured by western blot assay. RESULTS AND CONCLUSION:Mankin’s score was significantly lower in theTougu Xiaotong capsule group and Zhuanggu Guanjie Wan group compared with the model group (P < 0.01), in a time-dependent manner. Immunohistochemical staining indicated that the positive cels of urokinase-type plasminogen activator and urokinase-type plasminogen activator receptor were significantly decreased, but plasminogen activator inhibitor was significantly increased in theTougu Xiaotong capsule group andZhuanggu Guanjie Wangroup in a time-dependent manner. Western blot assay results had an identical trend to immunohistochemistry. These indicated thatTougu Xiaotong capsule showed preventive and therapeutic effects on osteoarthritis by regulating urokinase-type plasminogen activator system.

Objective To investigate the effects and mechanism of huoxuexiaoying tablet on experimental goiter of rats. Methods The rats were randomly divided into six groups: the control, the model control group, huoxuexiaoying tablet at different doses,and the sodium levothyroxine group ( Euthyrox group) . Except for the rats in the control,the rats in other groups were given with propylthiouracil (20 mg·kg-1 ·d-1 ) by intragastric ( i. g. ) administration every day for 60 days. Meanwhile, some rats were treated with huoxuexiaoying tablet at low (4. 4 g·kg-1·d-1),middle (8. 8 g·kg-1·d-1) and high dose (17. 6 g·kg-1 ·d-1 ) orally,and those in the Euthyrox group were given with 7. 8μg·kg-1 ·d-1 Euthyrox by i. g. administration. The rats in the control group were administrated with the same volume of saline (N. S). After 60 days of treatment,the rats were sacrificed,the organ indexes of thyroid and pituitary and the levels of free triiodothyronine ( FT3 )、free thyroxin ( FT4 ) and thyroid stimulating hormone(TSH) in serum,were tested. The expression of basic fibroblast growth factor(bFGF),transforming growth factor-β( TGF-β) and B-cell lymphoma 2 gene ( Bcl-2 ) were examined by immunofluorescence. Results Compared with the model,organ indexes of thyroid were significantly reduced by huoxuexiaoying tablet at three doses (P<0. 05),but not for the pituitary (P>0. 05). The levels of FT3 and FT4 were in a elevating trend,but TSH decreased with no significance (P>0. 05). The morphological structure of thyroid was greatly improved by huoxuexiaoying tablet in comparison with the model. In which the gland dilated,thyroid follicular restored to moderate size,epithelia were cubic or flattened and follicular cavity filled with abundant glial. The expression of bFGF and Bcl-2 decreased significantly (P<0. 01) while TGF-β expression increased notably (P<0. 01). Conclusion Huoxuexiaoying tablet has a great anti-goiter effect,the mechanism of which may be related to promoting thyroid cells apoptosis and inhibiting thyroid cells proliferation.

In this study, the mechanism by which Ad-p27mt inhibits the growth, invasion and metastasis of transplanted liver tumor was studied by examining the effects of Ad-27mt gene transfer on the expression of Bax, Bcl-2, VEGF and MMP-9 in the transplanted liver tumors in nude mice. The model of transplanted hepatic tumor was established in nude mice. The mice were then divided into three groups, which were injected with PBS, Ad-LacZ and Ad-p27mt and the growth of the transplanted liver tumor was observed. The expressions of P27, Bax and Bcl-2 proteins were detected by Western blotting and the expressions of VEGF and MMP-9 were immunohistochemically determined. Our result showed that the tumor size, expressions of Bax, Bcl-2 proteins, VEGF and MMP-9 were all lower than those in PBS and Ad-LacZ groups and the differences were statistically significant (P<0.05). Our study suggested that Ad-p27mt could inhibit the growth, invasion and metastasis of hepatic cancer by lowering the expressions of VEGF and MMP-9.

BACKGROUND:Previous studies have found that electroacupuncture can delay articular cartilage degeneration mediated by JAK-STAT signaling pathway through upregulating the expression level of transforming growth factor β1 as well as mRNA expression levels of STAT3, Smad3 and LepR. In the meanwhile, electroacupuncture can inhibit the mRNA expression of p38 and Fas mRNA mediated by MAPK signaling pathways, further inhibiting the apoptosis of chondrocytes. OBJECTIVE: To explore the effect of electroacupuncture on the degeneration of articular cartilage in rats with knee osteoarthritis based on Ras-Raf-MEK1/2-ERK1/2 signaling pathway. METHODS:120 male healthy Sprague-Dawley rats aged 2 months olds were selected and randomly divided into normal, model, 15-minite electroacupuncture and 30-minute electroacupuncture groups (n=30 per group). The rats in the latter three groups received the intra-articular injection of 4% papain bilaterally, and the remaining rats received no intervention. At 2 weeks after modeling, the latter two groups were respectively given 15- and 30-minute electroacupuncture, five times weekly for consecutive 12 weeks. The morphology of the cartilage was observed by hematoxylin-eosin staining, the expression level of interleukin-1β in the synovium was detected by ELISA assay, and the protein expression levels of Ras, Raf, MEK1/2, ERK1/2, C-MYC, C-FOS, and C-JUN were detected by western blot analysis. RESULTS AND CONCLUSION: Hematoxylin-eosin staining showed that: in the model group, the cartilage surface was rough, the cartilage layer became thinner, and the cartilage structure was damaged with incomplete tidal line; in the 15- and 30-minute electroacupuncture groups, the cartilage structure was complete with clear layers and complete tidal line. ELISA showed that the expression level of interleukin-1β in the model group was significantly higher than that in the normal group (P< 0.01), and the level in the 15- and 30-minute electroacupuncture groups was significantly lower than that in the model group (P < 0.05). Western blot assay found that compared with the normal group, the protein expression levels of Ras, Raf, MEK1/2, ERK1/2, C-MYC, C-FOS, and C-JUN were increased in the model group. However, all above protein levels except ERK1/2 in the 15- and 30-minute electroacupuncture groups were significantly lower than those in the model group (P < 0.01,P < 0.05). To conclude, electroacupuncture inhibits the degeneration of articular cartilage in osteoarthritisvia Ras-Raf-MEK1/2-ERK1/2 signaling pathway and downregulating the expression level of interleukin-1β.