Objective To establish a method for the determination of berberine hydrochloride in Weichangning granules.Methods RP-HPLC method was set up,using Merck-LiChrospher RP-18 column(4 mm ? 250 mm,5?m);the mobile phase was methnd-water(including 0.18 % phosphoric acid and 0.22 % sodium laurylsulfonate)(68 ∶ 32),flow rate being 1.0 mL / min;column temperature was 35 ℃ and the detection wavelength was 345nm.Results A good linearity was obtained in the range of 0.30 ~ 1.80 ?g(r = 0.9995).The average recovery was 98.32 %,with RSD 0.83 %.Conclusion The method is simple,rapid and with good reproducibility for the determination of berberine hydrochloride in Weichangning granules.

Objective To evaluate the long-term therapeutic effect of hyaluronic acid on adults with Kashia-Beck disease and the applicability of the joint dysfunction index.Methods A cluster-randomized,starch tablets-controlled trial was carried out depending on the joint dysfunction index of assessment for therapeutic efficacy on Kashin-Beck disease and self-assessment by patients of the Treatment Effect of Kaschin Beck Disease in Criterion.A total of 99 adults with Kashia-Beck disease were randomly divided into two groups:treatment group including 50 patients were intra-articular injected hyaluronic acid in knee weekly for 4 weeks,and the control group including 49 patients were treated with oral starch tablets for 3 months,2 tablets each time,3 times a day.We carried out 48 months follow-up and assessed the applicability of Kashia-Beck disease with treatment of hyaluronic acid.Results The improvement rates of joint dysfunction index were 80.0％ (40/50),83.0％ (39/47),86.4％ (38/44),85.7％(36/42),80.0％(36/45),and 47.1％(16/34) for 1 week and 1,2,3,6,48 months in treatment group,respectively,which were significantly higher than those of corresponding control group 25.0％ (12/48),28.3％ (13/46),33.3％ (16/48),25.0％ (10/40),30.0％ (12/40),and 21.4％ (6/28),and the differences were statistically significant between corresponding groups (x2=29.744,28.236,17.762,30.665,21.537,4.406; all P ＜ 0.05).Before treatment,the treatment group and control group in the joint dysfunction index scores were 7.59 ± 1.82 and 6.89 ± 1.97(t =1.837,P ＞ 0.05),respectively.After treatment,1 month follow up of the treatment group and control group in the joint dysfunction index scores were 3.44 ± 1.71 and 5.55 ± 2.34(t =4.972,P ＜ 0.01),respectively; 6 months follow up were 3.46 ± 2.78 and 6.10 ± 1.83(t =5.102,P ＜ 0.01),respectively; 48 months follow up were 5.13 ± 2.88 and 6.81 ± 3.07(t =2.219,P ＜ 0.05),respectively.The satisfaction of 85.1％ (40/47) in treatment group was significantly higher than that of 60.9％(28/46) in control group (Z =-4.012,P ＜ 0.01).Kappa value between joint dysfunction index and the overall evaluation of the treatment group after treatment was 0.526,which belong to moderate intensity of consistent degree (P ＜ 0.01).Conclusion The therapeutic effect of intra-articular injection of hyaluronic acid on adults with Kashia-Beck disease is significant and lasting,and the joint dysfunction index has better applicability to evaluate the long-term therapeutic effect.

Objective To evaluate the applicability of western Ontario and McMaster university osteoarthritis index (WOMAC) on Kaschin-Beck disease (KBD).Methods Clinical degree Ⅰ adult patients with KBD came from Yongshou and Linyou Counties in Shaanxi Province were investigated according to the historical diagnostic data and The Diagnosis Standard of Kashin-Beck Disease (GB 16003-1995),exclusion of other chronic diseases.Reliability of WOMAC was measured by retest reliability,1/2 coefficient and Cronbach'α reliability coefficient analysis; validity of the WOMAC was tested by the principal components,factor analysis and correlation analysis methods.Results Totally 200 adults patients with KBD were investigated,and 177 effective questionnaires were taken back (88.5 ％).Retest reliability was 0.754-0.853,1/2 coefficient was 0.886-0.971,and Cronbach'α reliability coefficient was 0.878-0.956.In three dimensions of WOMAC scale extracted a common factor,the cumulative variance contribution rate was 81.238％ through principal component factor.The pearson correlation coefficient between all items score of WOMAC and scale score and total score of WOMAC was more than 0.600.The differences of WOMAC score were not statistically significant in different ages,different grading of adult KBD patients (all P ＞ 0.05).Conclusion WOMAC scale used in KBD has good validity and reliability,but has low degree of differentiation.

Objective To investigate the improvement of quality of life of patient with Kashin-Beck disease (KBD) after knee joint cavity injection of hyaluronic acid.Methods Fifty KBD patients were selected in Yongshou County Shaanxi Province,and accepted knee joint cavity injection of hyaluronic acid (25 mg) for four times,one time a week.SF-36 health questionnaire was used to evaluate the quality of life of KBD patients,and the quality adjusted life years (QALYs) were calculated after 2 months.Results The total score of the quality of life of KBD patients was improved from 38.81 ± 17.39 to 49.35 ± 17.95 after knee joint cavity injection of hyaluronic acid.The scores of physiological function (PF),body pain (BP),general health (GH),energy (VT),social function (SF) and mental health (MH) in the eight dimensions of SF-36 scale were 54.42 ± 21.25,47.42 ± 20.80,48.00 ± 26.12,61.05 ± 19.14,68.09 ± 28.73 and 68.74 ± 14.85,respectively,after the treatment,which were higher than those before the treatment (41.51 ± 22.11,27.63 ± 11.78,38.76 ± 25.14,51.97 ± 18.14,57.89 ± 30.95 and 56.95 ± 20.47,t =-3.942,-6.344,-2.494,-2.785,-2.819 and-4.245,all P ＜ 0.05).The QALYs of KBD patients increased 0.051 ± 0.044 with 2 months followed up.Conclusion The quality of life (PF,BP,GH,VT,SF and MH) of KBD patients could be significantly improved after knee joint cavity injection of hyaluronic acid.

Objective To develop a new type of lumbar puncture needle to facilitate to measure intracranial pressure,decrease the risks for intracranial infection and brain hernia.Methods The needle was composed of a body,no.1 and no.2 sleeves,a stylet,a needle base,a catheter,joints and etc.The needle had body and stylet made of stainless steel,the catheter manufactured with medical silicone tube,the joints produced by medical rubber and the remained components by medical plastics.Results The developed needle executed pressure measuring,cerebrospinal fluid collection and medication injection with no extracting the stylet.The outflow velocity of cerebrospinal fluid was limited,and the incidences of the complications were decreased including infection,brain hernia and etc.Conclusion The lumbar puncture needle has simple structure,easy operation and high safety,and thus is worthy promoting practically.

Objective To analyze the effects of dengue virus 2 ( DENV-2 ) infection on the ex-pression of IL-29 in primary human umbilical vein endothelial cells ( HUVECs) cultured on hydrogel sub-strates .Methods Primary HUVECs were isolated and cultured on hydrogel substrates .DENV-2 stains were used to infect the primary HUVECs at a multiplicity of infection( MOI) of 10.Flow cytometry analysis was performed to detect the apoptosis and infection rate of HUVECs after 48 hours of culturing .The gene chip profiling was performed to analyze mRNA expression .The expression of IL-29 at mRNA and protein levels were measured by real-time fluorescent quantitative PCR analysis and double antibody sandwich ELISA as -say, respectively.Results Compared with 96.36%of baby hamster kidney (BHK) cells that were infected with DENV-2 stains, only 4.71%primary HUVECs cultured on hydrogel substrates were infected .The pri-mary HUVECs cultured on hydrogel substrates with or without DENV-2 infection showed no significant differ-ences with the rates of cell apoptosis and infection (P>0.05).A significant difference was observed with the expression of IL-29 at mRNA level between primary HUVECs cultured on hydrogel substrates and the cells cultured in plastic bottles (P<0.05).The results of the real-time quantitative PCR analysis and ELISA as-say showed that IL-29 was highly expressed in DENV-2 infected primary HUVECs cultured on hydrogel sub-strates as compared with those in control groups (P<0.05).Conclusion The expression of IL-29 was de-tected in DENV-2 infected primary HUVECs cultured on hydrogel substrates , which was significantly differ-ent from that in DENV-2 infected primary HUVECs cultured in plastic bottles .The successful establishment of hydrogel substrates as the model of vascular basement membranes might provide a new way for the investi -gation of the pathogenesis of DENV infection .

Objective To investigate the effects of substrate stiffness on the proliferation of human umbilical vein endothelial cells ( HUVEC) during dengue virus infection.Methods Polyacrylamide gels were prepared for cell culture [(0±4) kPa].The proliferation of HUVEC cultured on substrates with differ-ent stiffness was determined by using 3-(4,5-diethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfo-phenyl)-2H-etrazolium,inner salt ( MTS) assay.The cycle and apoptosis of HUVEC were determined by flow cytometry analysis.Dengue virus serotype 2 (DENV-2) strains were propagated and identified by con-ventional assays.The HUVEC were infected with DENV-2 strains at a MOI of 10 and cultured on traditional plastic and hydrogel substrates, respectively.The levels of nitric oxide (NO) and endothelin-1 (ET-1) were detected by nitric acid reductase assay and double antibody sandwich ELISA.Results Young′s modulus E value of the hydrogels was (3030 ±0.44) Pa.The proliferation of HUVEC and the expression of NO and ET-1 were enhanced along the increased substrate stiffness.However, no significant differences with the cell cycle and apoptosis were observed between cells cultured on different substrates.Conclusion The stiffness of substrates affected not only the proliferation of HUVEC, but also the release of cytokines during DENV-2 infection.The development of dengue fever was associated with the decreased secretion of vascular active substances as a result of blood vessel injury.The establishment of hydrogel substrates as the model of vascu-lar basement membranes might provide a new way for the in vitro investigation of the pathogenesis of DENV infection.

Objective To analyze the results of screening for tuberculosis (TB) in health examination participants and study the spontaneous occurrence of TB in the subjects with strong positive reaction to tuberculin skin test (TST) in ten years. Method Totally 12 598 health examination participants without past TB history were selected, of whom 8 896 were college students, 2 496 migrant workers and 1 206 had close contacts with active TB patients. They were screened by TST with strong positive reaction. All of subjects with TST strong positive results received chest X-ray examination and sputum acid-fast bacteria detection. The subjects diagnosed to have TB were given regularly anti-tuberculosis drugs treatment and followed up for ten years. The 429 subjects without TB and no isoniazid preventive treatment were chosen to be followed up for ten years and spontaneous occurrence of TB in first three years and the fourth to tenth years respectively, as well as the recurrence of TB for the patients who received anti-tuberculosis regimen were recorded. Result Thirty-seven cases were diagnosed as TB by TST screening, and the total detection rate was 0.29% (37/12 598). Among them 11 were college students (0.12%, 11/8 896), 12 were migrant workers (0.48%, 12/2 496) and 14 were close contacts (1.16%, 14/1 206) respectively. The detection rates were different among the three groups (χ2=31.40, P=0.000). Among 897 strong positive subjects, the strong positive rate was 7.12%(897/12 598), 316 were college students (3.55%, 316/8 896), 388 migrants workers (15.54%, 388/2 496), and 193 close contacts (16.00%, 193/1 206) respectively. There was significant difference in strong positive rate among the three groups (χ2=583.04, P=0.000), and the strong positive rate of college students was lower compared with that of the migrant workers and the close contacts (χ2=483.51 and 344.11, P<0.01). In ten years, 54 subjects were diagnosed as TB in 429 subjects with strong positive reaction to TST, the spontaneous cumulative incidence rate was 12.58% (54/429). Among them, the cumulative morbidity rate was 9.21%(14/152) in college students, 9.58%(18/188) in migrant workers and 24.72%(22/89) in close contacts respectively. The spontaneous morbidity rate of close contacts was higher than that of college students and migrant workers(χ2=10.63 and 11.21, P<0.001); 75 were lost in 398 participants, the dropout rate was 18.84%(75/398). In first three years of follow-up, 31 were diagnosed TB in 429 participants, the cumulative incidence rate was 7.23% (31/429). Of them 9 were college students (5.92%, 9/152), 10 migrant workers (5.32%, 10/188) and 12 close contacts (13.48%, 12/89) respectively (χ2=6.60, P=0.037). In the fourth to tenth years of follow-up, 23 were diagnosed TB in 398 participants, the cumulative incidence rate was 5.78% (23/398), which was not significantly different compared with the cumulative incidence rate of the first three years (χ2=2.50, P=0.37). Tirty-seven patients received standard anti-tuberculosis drug therapy for one year, no one of them had recurrence at ten-year follow-up. Conclusion The migrant workers and close contacts are the high-risk populations for TB. All of them with strong positive response to TST results are susceptible to TB. So regular physical examination is recommended for them and health management should be strengthened.

Objective To evaluate the reliability and validity of the Chinese version of European Five-dimensional Five-level Health Scale (EQ-5D-5L) for patients with Kaschin-Beck disease.Methods From May 2017 to September 2018,a total of 245 patients with Kaschin-Beck disease in Yongshou and Linyou counties,Shaanxi Province were interviewed face-to-face with the EQ-5D-5L.The health-related value score was used to calculate the overall health-related quality of life of patients with Kaschin-Beck disease.The higher the score,the better the overall health of the population,and vice versa (-0.391-1.000).The visual analogue system (VAS)score was used to calculate the individual health status,higher scores indicated that the health status of self-evaluation were better,and vice versa (0 to 100 points).Sixty-three patients were retested two weeks later.Evaluation was performed using internal consistency,test-retest reliability,structural validity and criterion validity.Results A total of 245 patients with Kaschin-Beck disease were investigated,including 127 males (51.8％) and 118 females (48.2％),aged (60.37 ± 7.10) years old.The medians and quartile ranges of EQ index and VAS of patients were 0.311 (0.504) and 42.5 (30.0) points,respectively.And the Cronbach coefficient of the five-dimensional score of the health description system was 0.828.The intra-class correlation coefficient (ICC) of each dimension of the health description system was 0.571-0.719.EQ index and VAS score showed that the two had a relatively stable correlation (rs =0.522,P ＜ 0.05),and the ICCs of EQ index and VAS score were 0.754 and 0.570,respectively;the EQ-5D-5L validity calculated by "Therapeutic Efficacy on Kaschin-Beck Disease" (WS/T 79-2011) was between 0.375-0.647 (P ＜ 0.05).Conclusions The overall health status of the Kaschin-Beck disease patient population and the health status of the individual self-evaluation measured by EQ-5D-5L are poor.The reliability test and validity test prove that the EQ-5D-5L is applied to the health status of patients with Kaschin-Beck disease.The EQ-5D-5L has good reliability and validity.

Objective To explore the main source of psychological stress for conscious patients in ICU. Methods A total of 40 awareness patients were investigated with the 24 h status-strait anxiety inventory ( STAI) and pittsburgh sleep quality index ( PSQI) for the sauce of psychological stress. Results Patients with different gender, occupation, marriage status, culture degree and department before transferred to ICU, their sleep quality scores were significantly different (P<0. 05). The result of STAI investigation had statistical significance compared with the norm in 24 h patients awaking (P<0. 05). The sleep quality of patients in ICU after waking for 24 h, compared with before they arrived ICU, there was a statistically significant difference (χ2 =34. 578,P<0. 01). For the influencing factors investigation of sleep quality, 95. 0% patients believed the sound pollution was the top reason impacted on sleep quality. Conclusions Sound pollution, safety and comfort are the main psychological stressors in the ICU area, which directly lead to different degrees of anxiety and are the key factors to affect the quality of sleep.