Objective:To study the feasibility of the determination of ephedrine hydrochloride in Tongxuan Lifei pills by acceler-ated solvent extraction ( ASE) combined with QuEChERS purification, and compare the results with those of the extraction method in Chinese Pharmacopoeia. Methods: The extraction efficiency of ephedrine hydrochloride in Tongxuan Lifei pills determined by HPLC was used as the evaluation index, and the operation parameters of ASE were optimized by orthogonal experiments. Results: The opti-mum conditions of ASE for the determination of ephedrine hydrochloride in Tongxuan Lifei pills were as follows:the samples were defat-ted by n-hexane ( the extraction temperature was 80℃, the static extraction time was 5 min for one cycle, and the flush volume was 100%. ) ,and then methanol was used as the extraction solvent for the extraction of ephedrine hydrochloride ( the extraction temperature was 80℃,the static extraction time was 8 min for three cycles) ,in the end, the impurities were purified by PSA purifying agent. Using the optimized ASE method to extract ephedrine hydrochloride in Tongxuan Lifei pills, the extraction time was reduced to 40 minutes with less interference, and compared with that of the method in Chinese Pharmacopoeia, the determination of the relative standard devi-ation was less than 5%. Conclusion:The ASE technique combined with QuEChERS purification is simple, quick and effective,and it can be used as the pretreatment method for the determination of ephedrine hydrochloride in Tongxuan Lifei pills.

Objective To explore the mechanism ofJiedu Qingfei Mixture for airway mucus hypersecretion of rat models with chronic obstructive pulmonary disease (COPD).Methods Airway instilling lipopolysaccharide combining fuming method was used to establish COPD models. Forty clean level Wistar strain rats were randomly divided into blank control group, model group,Jiedu Qingfei group, and clarithromycin group. Model group, Jiedu Qingfei group, and clarithromycin group were given normal saline,Jiedu Qingfei Mixture, and clarithromycin by gavage respectively, while the blank control group was raised normally for 30 d. All rats were killed on the 31st day for taking lung tissue (6 rats from each group were chosen randomly). Pathological changes of lung tissue and mucous glands hyperplasia were observed by HE staining method. NE and MUC5AC mRNA expression on lung tissue were detected by RT-PCR method. Protein expressions of NE and MUC5AC on pulmonary tissue and airway epithelium were detected by immunohistochemical method.Results Compared with blank control group, mucous glands hyperplasia on airway epithelium, mRNA expression of NE and MUC5AC in lung tissue, and protein expressions of NE and MUCA5C on airway epithelium in the model group significantly increased (P<0.05,P<0.01). Compared with model group, mucous glands hyperplasia on airway epithelium inJiedu Qingfei group significantly decreased (P<0.01), as same as clarithromycin group;Jiedu Qingfei group showed better effects on down-regulating NE and MUC5AC mRNA expression in lung tissue compared with clarithromycin group. MUC5AC protein expression on airway epithelium inJiedu Qingfei group significantly decreased (P<0.05), as same as clarithromycin group.Jiedu Qingfei group and clarithromycin group had no difference on NE protein expression in airway epithelium compared with model group.Conclusion Jiedu Qingfei group Mixture can reduce airway mucus hypersecretion of COPD by down-regulating MUC5AC expression through neutrophil elastase.

Objective To observe the effects of Qingjin Huatan Decoction on EGFR/MAPK signaling pathway of airway mucus hypersecretion rats with chronic obstructive pulmonary disease (COPD). Methods Intratracheal instillation of LPS combined with smudging method was used to establish COPD airway mucus hypersecretion rat models. Experimental rats were randomly divided into blank group, model group, Qingjin Huatan Decoction group and clarithromycin group. The blank group was normally fed, while the other three groups were given NS, Qingjin Huatan Decoction, and clarithromycin respectively for gavage, once a day for 30 days. All rats were killed on the 31st day, and pathological changes of lung tissue and mucous glands hyperplasia were observed by HE staining method. The gene expressions of EGFR and MUC5AC in lung tissue were detected by RT-PCR method. The protein expressions of P-EGFR, P-ERK, P-JNK, P-p38 and MUC5AC in pulmonary tissue and airway epithelium were detected by immunohistochemical method. Results Compared with the blank group, mucous glands hyperplasia on airway epithelium, protein expressions of P-EGFR, P-ERK, P-JNK, P-p38 and MUC5AC on airway epithelium significantly increased in the model group (P<0.01); gene expression of MUC5AC of lung tissue increased (P<0.05). Compared with the model group, mucous glands hyperplasia on airway epithelium, P-p38, P-ERK and MUC5AC protein expression on airway epithelium in Qingjin Huatan Decoction group significantly decreased (P<0.05, P<0.01); the protein expression of P-JNK increased significantly (P<0.01). EGFR and MUC5AC mRNA in lung tissue in Qingjin Huatan Decoction group decreased significantly (P<0.01). Conclusion Qingjin Huatan Decoction can reduce airway mucus hepersecrection of COPD by inhibiting ERK and p38 signal pathway on EGFR downstream.

Objective To evaluate factors influencing long-term survival of biliary atresia patients receiving Kasai procedure and the prevention and treatment of surgical complications.Methods Among 97 biliary atresia patients undergoing Kasai or Suruga operation from Mar 1989 to Mar 1999, 35 cases have survived over five years.These 35 cases were classified intraoperatively into type Ⅱ (4 cases) and type Ⅲ (31 cases).Procedures included hepaticojejunostomy (12 cases), Suruga operation (hepaticojejunostomy with a jejunal fistula in 21 cases), hepaticocystomy (2 cases).Six patients underwent second hepaticojejunostomy because of porta hepatis obstruction.Results (1) Eighteen patients suffered from early cholangiolitis, and four cases developed late cholangiolitis, 5-8 years after closing the ascending stoma.(2) Five cases presented upper digestive tract hemorrhage caused by portal hypertension.(3) All patients were followed up from 5 to 15 years with 2 deaths.Conclusion Conventional surgical procedures still take important part in treating biliary atresia before liver transplantation is popularized.Factors influencing long term survival of patients receiving Kasai′s operation include age at operation, pathological classification of extrahepatic bile duct, with or without cholangitis after operation, and portal hypertension.

Objective To conduct comparative study on the analgesic and anti-inflammatory effects as well as the acute toxicity of yunaconitine and 8-deacetyl-yunaconitine isolated from the processed products of Aconiti Knsnezoffii Radix.Methods The methods of hot plate test and writhing test were used to evaluate the analgesic effect. Anti-inflammation action was observed by the models of auricle swelling caused by dimethylbenzene. LD50 was determined by the method of Bliss.Results Yunaconitine and 8-deacetyl-yunaconitine have analgesia effect on the pain caused by hot-plate, but there were no statistically significant difference. The pain caused by acetic acid had obvious analgesic action. High and low dose of yunaconitine could significantly reduce the number of mice body torsion and extend the incubation period of pain in mice. The effect of 8-deacetyl-yunaconitine was remarkable only in the high dose. Compared with solvent group, there were little differences in inhibiting effect of auricle swelling caused by dimethylbenzene, and anti-inflammatory action was not exact. The poisonousness of yunaconitine was nearly 20 times of 8-deacetyl-yunaconitine.Conclusion Yunaconitine and 8-deacetyl-yunaconitine may be the analgesic medicine for peripheral analgesic effect. The poisonousness of 8-deacetyl-yunaconitine is less than yunaconitine, the effect is remarkable to the pain caused by acetic acid, and the security is high.

Objective To establish and evaluate a method for determination of total arsenic in urine by test-tube rapid digestion hydride generation atomic fluorescence spectrometry.Methods After digestion of urine samples using graduated test-tube and graphite digestion apparatus,arsenic content in urine was determined with atomic fluorescence spectrometer.Then the test results were evaluated by using quality control measures,such as precision and accuracy experiments,and the results between different laboratories were reviewed and compared.Results The urinary arsenic was in a linear range of 0-0.300 mg/L,correlation coefficient (r) ＞ 0.999 3,detection limit was 0.000 21 mg/L,relative standard deviation (RSD) ≤4.62％ and the recoveries of standard addition were 93.9％-104.3％.The value of standard reference material measured was within the allowable range.The blind sample of the national urinary arsenic was qualified.Conclusions This method is suitable for large scale determination of urinary arsenic for its micro sample amount needed,less interference and strong practicability.The error results are in a controlled range.

Objective To study the process conditions of purifying the total flavonoids from Cacumen Platycladi by AB-8 resin. Methods Spectrophotometric method was used to detect the total flavonoids,and HPLC was used to determine quercitrin and analyze the characteristic peaks of Fingerprint. Results The purification effect was satisfactory when the concentration of original solution of the extract of Cacumen Platycladi was 0.20 g?mL-1,the loading amount was 0.375 g of Cacumen Platycladi per 1 mL of wet AB-8 resin,the adsorption velocity was 1 mL?min-1,the eluant was 70 %alcohol being 4 times as much as the resin volume,and the elution velocity was 2 mL?min-1. AB-8 resin could be used for 3 times repeatedly after being reproduced by 95 %of ethanol and 1 moL?L-1 of natrium hydroxydatum (NaOH). The remaining rate of the total flavonoids and quercitrin was over 70 %and 95 %respectively,and the remaining rate of peaks 1～3 was over 90 %. After being purified by the B-8 resin,the contents of the total flavonoids and quercitrin were raised 2.48 times and 3.29 times more than those before purification respectively. Conclusion AB-8 resin is fit for separating and purifying the total flavonoids from Cacumen Platycladi.

Objective To explore the effect of gravitational traction on height change of degenerative disc of New Zealand white rabbit.Methods In order to create intervertebral disc degeneration models, the right front side of the annulus fibrosus of lumbar vertebrae of rabbits were damaged by 16-gauge needle. After confirming the success of modeling, 20 animal models were randomized into 2 groups, 15 of which received a routine gravitational traction using a model of our own design, 30 min per day, for 8 weeks, while the control group received no treatment at all. Radiographic exam were performed in the 2nd, 4th, 6th and 8th week, after intervention and the disc height index percent (DHI%). Results Compared to the baseline, DHI% increased significantly 2 and 4 weeks after gravitational traction (P<0.05), and there was no further effect as the traction continued. However, compared to the control group, an improvement of DHI% was found on all the time points in the gravitational traction group (P<0.001). Conclusion Gravitational traction may delay and inhibit intervertebral disc degeneration by increasing height of degenerated disc of New Zealand white rabbit.

Objective To explore the effects of nerve growth factor (NGF) and low frequency electricity stimulus on the configurations of skeletal muscle cells and the change of muscle fibre types in the denervated skeletal muscle separately.Methods The denervated rat model was established and model animals were injected with the NGF and given the stimulus (frequency=2 Hz) about 30 days. The configurations and the change of muscle fibre types were observed by immunohistochemistry and image analysis.Results The muscle fibre was in chaos and the boundary was not obvious among cells in the denervated rats; the muscle fibre of the denervated rats with NGF injection and low frequency electricity stimulus was more regular and the boundary of cells was clearer, the cells number was more than those of the denervated rats. Compared to normal rats, the proportion of Ⅰ muscle fibre in the denervated rats increased ( P<0.05), whereas the proportion of Ⅱ muscle fibre decreased ( P<0.05); it had no significant differences of the two types of muscle fibre between the denervated rats with NGF injection, low frequency electricity stimulus and the denervated rats ( P<0.05).Conclusion NGF injection and low frequency electricity stimulus can make the configurations of denervated muscle to better.

Objective To find out the distribution of drinking-tea-borne fluorosis in the six ethnics in Qinghai Province, and to provide basic data for prevention and control of the disease. Methods In 2010, according to the requirement of “The National Surveillance Program of Drinking-Tea-borne Fluorosis”, six ethnics accounted for 99.59% of total population in Qinghai Province were investigated in 28 counties having brick-tea drinking habit. Three townships and a town in each county, two administrative villages(residents’ committee) in each township and town were chosen and 50 adults in each administrative village and residents ’ committee were selected to check skeletal fluorosis, dental fluorosis, urine fluoride and daily drinking amount of tea water. Five to six samples of drinking tea water were determined. Dental fluorosis was examined by Deans method; the fluoride content of brick-tea and urine were determined by fluoride ion selective electrode; the skeletal fluorosis was diagnosed based on “Endemic Osteofluorosis Clinical Indexing Diagnosis Standard”( WS 192-2008 ) . Results A total of 10 335 adults were surveyed, the number of Tibetan, Han, Hui, Mongolian, Tu and Salar ethnics were 4 972, 3 063, 1 196, 634, 235 and 235, respectively. The daily drinking amounts of tea water in Mongolian, Tibetan, Hui, Tu, Han and Salar ethnics were 2.53, 2.19, 1.74, 1.63, 1.22 and 1.07 L, respectively. Daily fluoride intakes in Tibetan, Mongolian, Tu, Hui, Han and Salar ethnics were 3.99, 2.78,2.27, 2.16, 1.78 and 1.28 mg, respectively. The medians of urinary fluoride concentration of the Tibetan, Tu, Hui, Han, Mongolian and Salar ethnics were 1.46, 1.19, 1.12, 0.98, 0.93 and 0.81 mg/L, respectively. The prevalence rates of dental fluorosis of the Hui, Han, Tibetan, Tu, Mongolian and Salar ethnics were 34.53%(413/1 196), 27.07%(829/3 063), 21.60%(1 074/4 972), 20.00%(47/235), 17.98%(114/634) and 6.38%(15/235), respectively. The incidence rates of clinical skeletal fluorosis of the Tibetan, Mongolian, Han, Hui, Tu and Salar ethnics were 13.42%(667/4 972), 11.04%(70/634), 9.31%(285/3 063), 7.61%(91/1 196), 5.53%(13/235) and 4.26%(10/235), respectively. Conclusions The distribution and prevalent status of drinking-tea-borne fluorosis in the six ethnics of Qinghai Province are different. Tibetan and Mongolian ethnics are the key population concerning the prevention and control of the disease.