AIM: To establish the quality standard for Chanhua Junsitifen Capsule(Paecilomyces cicadae(Miquel) samson). METHODS: Amino acids,adenine,adenosine,uridnine in Chanhua Junsitifen Capsule were identified by TLC and adenosine was determined by HPLC.Chromatographic conditions were adopted as follows:co-lumn: Inertsil C_18(4.6 mm?250 mm,5 ?m),mobile phase: MeOH-0.05 mol/L KH_2PO_4 solution(15∶85),wavelength: 260 nm,flow rate: 1.0 mL/min. RESULTS: The average recovery was 99.60 % and RSD was 0.62 % and the linear range of adenosine was in the range 0.007 475-2.99 ?g. CONCLUSION: This method is simple,rapid with a good reproducibility.This method can be used for the quality control of Chanhua Junsitifen Capsule.

The single molecule imaging and technologies that developed in 1990 s have successfully probed the dynamics of single molecule enzyme catalysis in real time in vitro. Ever since then, single molecule enzymology has entered the golden age of rapid developing. Individual features of each enzyme hidden in the overall average have been discovered, and many new catalytic mechanisms have been proposed. Single molecule enzymology sheds light on the dynamic interactions between enzymes and substrates or products, deepening the understanding of biochemical reactions. This review described the recent research progresses of single molecule protease and ribozyme.

ObjectiveTo observe the clinical efficacy ofTong Du Tiao Shen(unblocking the Governor Vessel and regulating spirit) needling method plus Nimodipine in treating vascular dementia.MethodSixty patients with vascular dementia were randomized into a treatment group and a control group, 30 in each group. The control group was intervened by oral administrationof Nimodipine, while the treatment group was additionally intervened byTong Du Tiao Shenneedling method. Two weeks were taken asa treatment course, and the intervention lasted 2 courses. The Mini-Mental State Examination (MMSE) and Activities of Daily Life (ADL) were evaluated and compared before and after intervention, and the therapeutic efficacy was determined.Result After intervention, there were significant differences in comparing MMSE and ADL scores between the two groups (P<0.05). ConclusionTong Du Tiao Shenneedling method plus Nimodipine is effective in treating vascular dementia, and worth promotion in clinic.

How to identify active constituents of traditional Chinese medicines (TCMs) and study their interactions are key problems in the development of TCMs. The inhibitory effect of six alkaloids from Rhizoma Coptidis (RC) on Shigella dysenteriae (S. dysenteria) growth had been investigated by microcalorimetry in this study. Main active constituents of RC were confirmed by comparing their contributions to the bacteriostatic effect, and the interactions among active constituents were further researched. According to the result, in 0.8 mg-mL-1 extract of RC, the contributions of six active alkaloids including berberine, coptisine, epiberberine, palmatine and the combination of jatrorrhizine and columbamine were 52.83%, 36.31%, 2.49%, 4.27% and 3.21%, respectively. Therefore, berberine and coptisine were the main active constituents of RC that inhibited the growth of S. dysenteria. The study of interactions among the six alkaloids indicated that, 1 there were some contstituents antagonizing the inhibitory effect of RC, 2 there was a synergy effect between berberine and coptisine, 3 there were additive effects between other four alkaloids and the main active constituents. These results may provide some useful references for the establishment of the quality standard for RC and the development of multi-component TCMs.

Objective:To investigate the role of reducing resistance and distraction in rapid teeth movement and its reliability by establishing the Beagle dogs’ experimental model.
Methods:The left or right sides in mandibles of 20 beagles were randomly operated with different treatments:distraction twice a day through reducing resistance;distraction 6 times a day through reducing resistance;conventional distraction through reducing resistance;and conventional distraction (the control group). Each treatment was carried out in 10 sides. The pulp vitality, tooth mobility and distance of teeth transportation were evaluated at different time points:before the distraction, distraction after 15 days, retaining 30 days after 15 days of distraction. The degree of inclination, root resorption and alveolar bone density of the compressive areas were evaluated by cone-beam computed tomography images.
Results:The distance of teeth transportation was similar in groups distraction twice daily and 6 times a day through reducing resistance (P>0.05), but their speed of transportation was significantly higher than that of conventional distraction through reducing resistance. The conventional distraction group had the lowest speed of transportation. The pulp vitality of distracted teeth was normal, and no root comprehensive resorption and periodontal defect were found. Distracted teeth in the reduced resistance and distraction groups (13.9°±3.5°) tipped more that in the conventional distraction group (6.6°±1.3°) (P<0.05).
Conclusion:Reducing resistance and distraction are inseparable factors to realize fast teeth moving. The rate of orthodontic tooth movement can be accelerated through resistance reduction and periodontal distraction without obvious unfavorable effects but at minimal acceptable teeth inclination.

This study aims at trying to establish a novel method of sterility test for injections based on biothermodynamics, in order to overcome the deficiencies of routine sterility tests such as long detecting cycle, low sensitivity and prone to misjudgments. A biothermodynamics method was adopted to rapidly detect the microorganism contamination of injections by monitoring the heat metabolism during the growth of microbe. The growth rate equal to or greater than zero and the heat power difference of P(i) and P(0) with three folds higher than the noise of baseline were chosen as indexes to study the heat change rule of microbe. In this way, the effectiveness of the new method to detect strains required by conventional sterility test or in injection samples was also investigated. Results showed that the Gram-positive bacteria, Gram-negative bacteria and fungi demanded by sterility testing methodology could be detected by biothermodynamics method within 10 hours, with the sensitivity lower than 100 CFU x mL(-1). Meanwhile, this method was successfully applied to the sterility test of Compound Yinchen injection (FFYC), Shuanghuanglian powder injection (SHL) and Compound Triamcinolone injection (TAND) which were sterilized with different degrees. Therefore, the biothermodynamics method, with advantages of fast detection and high sensitivity, could be a complementary solution for conventional sterility tests.

The study is to report the establishment of a method of screening the antitumor compounds based on the dynamic bio-response profile of cells to make up for the shortages of conventional end-point tests such as tedious operation and low sensitivity. Based on the principle of electric impedance of cells, the real-time cell electronic sensing (RT-CES) system was used to monitor the effect of epirubicin (EPI), cisplatinum (DDP) and carboplatin (CBP) on the growth of HepG2 cells, with the cell index (CI), half maximal inhibitory concentration (IC50) and detachment curve as evaluation indexes. Meanwhile, cell counting kit-8 (CCK-8) and microscopy were applied for verification. The results showed that CI curve could sensitively real-time profile the inhibitory effect of model drugs on HepG2 cells. The IC50 of EPI, DDP and CBP were 0.53 +/- 0.04, 9.79 +/- 0.26 and 597.00 +/- 3.79 microg x mL(-1), respectively. What's more, the significant differences of detachment curves of the three drugs indicated that their functional mechanisms might be different, this is consistent with the literature. The RT-CES system with non-invasive, label-free and real-time characteristics could be used to monitor the bio-response profile of the three drugs to HepG2 cells, allowing to qualitatively and quantitatively distinguish the antitumor activities of the three drugs, and could be a complementary method for the present screening of antitumor compounds.

Objective To investigate the effect and mechanism of colchicine on mouse liver cancer via Hippo sig-naling pathway.Methods The 6-week-old male C57BL/6J mice were randomly divided into 3 groups:diethylni-trosamine(DEN)/carbon tetrachloride(CCl4)/ethanol(C2H5OH)induced mouse liver cancer model and col-chicine(0.1 mg/kg)intervention were established in control group,model group and colchicine group.From week 1st to week 2nd,the model group and the colchicine group were intraperitoneally injected with 1.0％DEN once a week.From week 3rd to week 7th,20％CCl4 dissolved in olive oil solution(5 ml/kg)was intragastric ad-ministration twice a week.From week 8th to week 18th,20％CC14 dissolved in olive oil solution(6 ml/kg)was intragastric twice a week.The colchicine group was given continuous intragastric administration for 20 weeks.The control group was given the corresponding solvent.Liver index,alanine aminotransferase(ALT)and aspartate ami-notransferase(AST)serum biochemical indexes were detected.Western blot and immunofluorescence were used to detect the expression levels of MST1,pYAP,YAP,pTAZ and TAZ proteins in liver tissues of mice in each group.Results The liver surface of mice in the control group was smooth and soft,while the liver of mice in the model group was rough and hard with granular nodules.The above lesions were significantly improved in the colchicine group.HE staining showed that the liver lobular structure of mice in the control group was normal,while the liver lobular structure of mice in the model group was disordered,with a small amount of fat droplets,extensive tissue necrosis,inflammatory cell infiltration,and fat vacuoles.The degree of liver lesions of mice after colchicine inter-vention was significantly reduced.The results of immunofluorescence and Western blot showed that compared with the control group,the protein expression levels of pYAP and pTAZ in liver tissue of model group mice were signifi-cantly decreased,while the protein expression levels of MST1,YAP and TAZ increased.After colchicine interven-tion,the protein expression levels of MST1,pYAP and pTAZ were significantly up-regulated,while the protein ex-pression levels of YAP and TAZ were down-regulated.Conclusion The therapeutic effect of colchicine on mouse liver cancer may be related to its activated Hippo signaling pathway.

Objective:To investigate the epidemiology of human papillomavirus (HPV) infection and gene subtypes among women in Qingdao form 2015 to 2019, so as to provide scientific support for the prevention and treatment of HPV related cervical cancer in Qingdao.Methods:A total of 26396 female cervical exfoliated cell specimens were collected at the Affiliated Hospital of Qingdao University from January 2015 to October 2019, then 17 high-risk HPV genotypes and 6 low-risk HPV genotypes were detected by PCR and reverse dot blot in vitro, and prevalence and year, genotype and age distribution were analyzed.Results:The total prevalence of HPV was 18.17%. The prevalence of single infection, double infection and multiple infection were 13.41%, 3.40% and 1.35% respectively. The infection rate of 7 high-risk subtypes (HPV52, 16, 58, 53, 56, 51, 68) and 3 low-risk subtypes (HPV81, 42, 43) exceeded 1%. The HPV prevalence of different age groups was significantly different and the prevalence showed a bimodal pattern among different age groups. There were two peaks in the ≤25 years group and 46-50 years group, which were 23.61% and 19.63% respectively. There was no significant difference in genotype distribution among different age groups.Conclusions:The cervical HPV infection rate of women in Qingdao is at a medium level, and single infection is the main pattern in different genotypes and age groups. In addition to the vaccine coverage subtypes, the prevalence of other subtypes such as HPV 53, 56, 51 and 68 in this region is also high. Therefore, regular HPV screening is still needed after vaccination to find out HPV genotypes which are not covered by the vaccine. It is suggested that the government optimize the HPV vaccine coverage genotypes to prevent the occurrence of HPV infection associated with cervical cancer.

Objective To comparatively observe the value of TIPS combined with catheter-directed thrombolysis and percutaneous transhepatic portal vein catheterization thrombolysis for acute non-cirrhotic non-neoplastic portal vein thrombosis(PVT).Methods Twenty-five patients with acute non-cirrhotic non-neoplastic PVT were retrospectively enrolled and clustered into TIPS group(n=17,underwent TIPS combined with catheter-directed thrombolysis)and liver puncture group(n=8,underwent percutaneous transhepatic portal vein catheterization thrombolysis)according to the access of thrombolysis.The technical success rate,duration of catheter-directed thrombolysis,complications within 7 days,as well as portal vein patency 3 months after treatment,Child-Pugh grading of liver function and occurrence of hepatic encephalopathy(HE)were recorded and compared between groups.Results The technical success rates were both 100%in 2 groups.There was no significant difference of the duration of catheter-directed thrombolysis between groups(P＞0.05).The thrombolytic effect in TIPS group was better than that in liver puncture group(P＜0.05).No significant difference of the occurrence of bleeding within 7 days was found between groups(P＞0.05).After 3 months'follow-up,the degree of portal vein patency in TIPS group was higher than that in liver puncture group(P＜0.05).No significant difference of Child-Pugh grading of liver function nor occurrence of HE was found between groups(both P＞0.05).Conclusion Both TIPS combined with catheter-directed thrombolysis and percutaneous transhepatic portal vein catheterization thrombolysis were effective for treating acute non-cirrhotic non-neoplastic PVT,and the thrombolytic effect of the former was better than the latter.