Objective:To observe the efficacy of Qingre Lishi prescription in treating children with acute bacterial lower urinary tract infection of bladder damp-heat syndrome， and to explore its mechanism of action. Method:Eighty children with acute bacterial lower urinary tract infection of late bladder damp-heat syndrome who were admitted to the Affiliated Hospital of Changchun University of Chinese Medicine were divided into control group and observation group， 40 cases in each group. Patients in control group were given Bazhengsan for oral treatment on basis of basic treatment， while patients in observation group were given Qingre Lishi prescription for oral administration plus external washing treatment. After two weeks of treatment， the clinical and etiological effect， traditional Chinese medicine （TCM） syndrome scores， antipyretic time and urinary negative time， adverse reactions， and urine pathogens （Escherichia coli， Enterococcus faecalis， Strange proteus， Klebsiella pneumoniae）， serum inflammatory factor indicators ［tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， calcium lowering PCT， white blood cell count （WBC） and serum C-reactive protein （CRP）］， immune function indicators ［T cell subsets （CD3⁺， CD4⁺， CD8⁺） and complement （C3， C4）］ were comapred between two groups. Result:The clinical efficacy of observation group was 92.50% （37/40）， which was significantly higher than 65.00% （26/40） in control group （χ²=9.038， P<0.01）， the etiological efficacy of observation group was 87.50% （35/40）， which was significantly higher than 60.00% （24/40） in control group （χ²=7.813， P<0.01）. After treatment， the scores of TCM syndromes of the two groups were significantly reduced （P<0.05）. The scores of fever， frequent urination， urgent urination， painful urination， difficulty urinating and abdominal pain in two groups were significantly lower than those before treatment （P<0.05）， and the TCM syndrome scores in observation group were lower than those in control group （P<0.05）， the antipyretic time and urinary bacteria turning negative time of observation group were significantly lower than those in control group （P<0.05）， the Escherichia coli， Enterococcus faecalis， Proteus mirabilis， Klebsiella pneumoniae pathogenic bacteria detected in both groups were both significantly lower than those before treatment （P<0.05）. After treatment， the levels of inflammatory factors such as TNF-α， IL-6， IL-8， PCT， WBC and CRP in two groups were significantly lower than those before treatment （P<0.05）， the immune function of the two groups was significantly improved， and the levels of CD3⁺， CD4⁺， C3， and C4 in observation group were higher than those in control group（P<0.05）， and the CD8⁺ level was lower than that in control group （P<0.05）. The incidence of adverse reactions had no significant difference between two groups. Conclusion:Qingre Lishi prescription has good clinical effect in treating children with acute bacterial lower urinary tract infection with bladder damp-heat syndrome. It can improve TCM syndromes and clinical symptoms. Its mechanism is related to inhibiting pathogenic bacteria， reducing inflammation， and improving immune function， and it has good security.

Objective:To study the protective effect and mechanism of Ranae Oviductus protein hydrolysate （ROPH） on the expression of pathway-related proteins in ethanol-induced L-02 cell injury. Method:The ROPH was prepared by compound enzymatic hydrolysis. L-02 cell injury model was induced with 400 mmol·L^-1 ethanol. Cell viability was detected by cell counting kit-8 （CCK-8） assay. Cell cycle and apoptosis were examined by flow cytometry. JC-1/Hochest staining was employed for qualitative investigation. The expression of related proteins in apoptosis， mitogen-activated protein kinase （MAPK） signaling pathway， and pyroptosis in L-02 cells was detected by Western blot. Result:The results of the CCK-8 assay showed that 400 mmol·L^-1 ethanol could induce L-02 cell injury within 12 hours. Compared with the blank group， the model group showed decreased viability of L-02 cells （P<0.01）， elevated percentage of the cell cycle in the G₀/G₁ phase （P<0.01）， increased total cell apoptosis rate （P<0.01）， reduced mitochondrial membrane potential （P<0.01）， up-regulated expression of apoptosis-related proteins ［B-cell lymphoma-2 （Bcl-2）-associated X protein （Bax）， Cytochrome C （Cyt C）， and cysteine-dependent aspartate specific protease-3 （Caspase-3）］ （P<0.05， P<0.01） and MAPK signaling pathway-related proteins ［C-Jun amino-terminal kinase （JNK） and p38 MAPK］ （P<0.05， P<0.01）， and potentiated expression of pyrolysis-related proteins Caspase-1 and interleukin-1β （IL-1β） （P<0.05）. Compared with the model group， the ROPH treatment group exhibited improved cell cycle arrest （P<0.05， P<0.01）， diminished total cell apoptosis rate （P<0.01）， elevated mitochondrial membrane potential in a dose-dependent manner， down-regulated expression of Bax， Cyt C， and Caspase-3 proteins （P<0.05， P<0.01）， up-regulated expression of Bcl-2 protein （P<0.05， P<0.01）， and a downward trend in expression of proteins related to MAPK signaling pathway and pyrolysis （P<0.05， P<0.01）. Conclusion:ROPH could inhibit oxidative stress-triggered liver injury in ethanol-induced cells by improving mitochondrial membrane potential， reducing the expression of proteins in the mitochondria-mediated apoptosis pathway， and inhibiting the expression of proteins related to the MAPK signaling pathway and pyrolysis pathway to reduce the mitochondrial dysfunction and inflammatory response in ethanol-induced L-02 liver cells and inhibit oxidative stress， thereby exerting a therapeutic role in alcoholic liver injury.

OBJECTIVE：To study the effects of gin seng root extract on autophagy ，proliferation and cytokine of splenic lymphocyte of mice ，and to study its mechanism. METHODS ：The splenic lymphocyte of mice were divided into blank control group，positive control group （concanavalin A ，5 μg/mL），different concentration of ginseng root extract groups （32，125，500 μg/mL，by lyophilized powder ）. After 48 h of culture with the corresponding medicine ，acridine orange staining method was used to detect autophagy of splenic lymphocyte ；CCK-8 assay was used to detect the cell proliferation ；ELISA assay was used to determine the levels of IL- 4，IL-6 and TNF-α in cell culture；Western blotting method was used to detect the expression of LC 3B and Beclin- 1，as well as the phosphorylation of AMPK ，AKT and mTOR. RESULTS ：Compared with blank control group ，32， 125，500 μg/mL ginseng root extract could increase the number of acidic autophagosomes in splenic lymphocytes of mice（P＜0.05 or P＜0.01）；125，500 μg/mL Ginseng root extract could significantly enhance the survival rate of splenic lymphocytes，the levels of IL- 4，IL-6 and TNF-α and the transformation of LC3BⅠ to LC 3BⅡ，significantly increased the protein expression of Beclin- 1， quinacrine cationic liposomes for treating non-small cell @163.com lung cancer[J]. J Drug Target ，2015，23（3）：232-243. the phosphorylation of AMPK protein ，and significantly reduced the phosphorylation level of AKT and mTOR proteins ，with statistical significance （P＜0.05 or P＜0.01）. CONCLUSIONS ：Ginseng root extract can induce autophagy of mice splenic lymphocytes，activate the activity of splenic lymphocytes ，regulate the secretion of cytokine by activating AMPK and inhibiting the activation of AKT which can inhibit the activity of mTOR ，so as to exert immune enhancement effect.

ObjectiveTo discuss the characteristics of psychological assistance hotline calls and operators' coping strategies of before and after the COVID-19 outbreak， in order to further improve the assistance ability of the psychological crisis hotline. MethodsA retrospective study was conducted on the demographics characteristics， call problems， coping strategies， and call time trends recorded by Changchun psychological assistance hotline information registration platform before the epidemic in Changchun City （January 20， 2019-April 20， 2019） and during the epidemic period （January 20， 2020-April 20， 2020）. ResultsThe differences between gender， age， marital status， location， and occupation type before and during the epidemic were statistically significant （χ²=11.205， 234.240， 152.083， 265.458， 353.385， P<0.01）. The number of different help calls had a statistically significant difference before and during the epidemic （χ²=185.088，P<0.01）. The difference in the number of operators’ different coping strategies before and during the epidemic was statistically significant （χ²=226.810， P<0.01）. Before the epidemic， the main peak of incoming calls was concentrated at 16∶00 to 17∶00， and the secondary peak was concentrated at 22∶00 to 23∶00. During the epidemic， the main peak of incoming calls was also concentrated at 16∶00 to 17∶00， while the secondary peak was concentrated at 10∶00 to 11∶00. ConclusionDuring the COVID-19 epidemic， the number of calls to the psychological assistance hotline was higher than that before the outbreak. The main peak time for calls was the same， and the secondary peak was adjusted from 22∶00 to 23∶00 to 10∶00 to 11∶00. During the epidemic， the number of calls from male， 30 to 39 years old， married， local and staff in Changchun was the most， psychological problems counseling and operator referral strategy were the most before and after the epidemic.

Animals ; Biomarkers ; analysis ; metabolism ; Carcinoma, Hepatocellular ; diagnosis ; metabolism ; Chromatography, High Pressure Liquid ; methods ; Discriminant Analysis ; Fatty Liver ; diagnosis ; metabolism ; Humans ; Liver Failure ; diagnosis ; metabolism ; Liver Neoplasms ; diagnosis ; metabolism ; Mass Spectrometry ; methods ; Metabolomics

OBJECTIVE：To establish the f ingerprint of Qinlian runfei decoction，determine the contents of 11 components， and conduct cluster analysis and orthogonal partial least squares discriminant analysis （OPLS-DA）. METHODS ：HPLC method was used. The determination was performed on ZORBAX Eclipse Plus C 18 column with mobile phase consisted of acetonitrile- 0.05％ phosphoric acid solution （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelength was set at 260 nm， and column temperature was 30 ℃. The sample size was 10 μL. Using wogonoside as reference，HPLC fingerprints of 10 batches of Qinlian runfei decoction were drawn and the similarity evaluation was conducted with Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition），the common peaks were also confirmed ；the contents of 11 components in Qinlian runfei decoction were determined by the same method. SPSS 21.0 software was used for clus ter analysis ，and SIMCA 14.0 software was used for OPLS-DA to screen marker components affecting quality. RESULTS ：There were 21 common peaks in 10 batches of Qinlian runfei decoction ，and the similarity with control fingerprint was greater than 0.98. A total of 11 common peaks were identified ， which were rutin ， forsythiaside A ， forsythiaside B ， iris， irigenin， baicalin， forsythiaside， wogonoside， baicalein， irisflorentin and wogonin. The line ar ranges of 11 components were 9.960 0-49.800 0，1.974 0-9.870 0，0.672 0-3.360 0，0.960 0-4.800 0，0.549 0- 2.745 0，5.040 0-25.200 0，1.374 0-6.870 0，0.615 0-3.075 0，0.759 9-3.795 0，0.162 0-0.810 0，0.042 0-0.210 0 μg（all r＞ 0.999）； RSDs of precision ， stability （48 h） and repeatability tests were less than 2％ ； the average recoveries were 95.81％-100.29％ with RSDs of 0.43％-1.73％（n＝6）；the contents were 8.924 4-12.820 8，0.352 2-0.868 7，0.435 6-0.711 2， 0.389 8-1.309 0，0.335 8-0.530 1，1.680 5-4.542 3，0.701 8-1.584 2，2.240 2-5.442 5，2.351 0-5.558 9，0.106 0-0.182 2，0.076 8- 0.128 9 mg/g，respectively. The results of cluster analysis showed that when class spacing was 10，it could be divided into two groups，S1-S3 and S 4-S10；when the class spacing was 5，the second class could be divided into two categories ，S6，S7，S9 were clustered into one category ，and S 4，S5，S8，S10 were clustered into one category. The results of OPLS-DA analysis showed that S6，S7 and S 9 were at the top of the figure ，S4，S5，S8 and S 10 were at the lower left side of the figure ，and S 1-S3 were at the lower right side of the figure ，which was consistent with the cluster analysis results ；VIP values of baicalin ，iris，forsythiaside A ， baicalein and wogonoside were all greater than 1. CONCLUSIONS ：Established fingerprint and content determination methods have high precision and good stability. Combined with multivariate statistical analysis ，it can be used for the quality control of Qinlian runfei decoction. Five components as baicalin are the marker components affecting the quality of Qinlian runfei decoction.

The use of animal medicine has a long history in China, it has the characteristics of high curative effect,strong activity, wide application and great potential. However,the circulation of animal medicine in current market mixed counterfeit variety and complex. Molecular identification technology of DNA barcoding is an emerging molecular biotechnology in recent years, it is a powerful supplement to traditional identification methods. This method can well identify animal species at the molecular level and has high accuracy, it can identify animal medicines quickly and monitor the medicine market effectively. This article summarizes the research process of molecular identification of DNA barcoding, the application of DNA barcoding in medicinal animals identification in recent years, and the limitations of DNA barcoding technology.
Animals ; China ; DNA ; DNA Barcoding, Taxonomic ; Research

OBJECTIVE：To optimi ze the optimal composition proportion of 4 ingredients （Panax ginseng ，Astragalus membranaceus，Polygonatum sibiricum ，Lycium chinensis ）in Compound ginseng immune-enhancing formula （CGIF），and to study immune activity and acute toxicity of the extracts with the optimal ratio. METHODS ：The cell activity test was used to screen the crude drug concentration range of 4 ingredients. After treated with different crude drug concentrations of each medicinal material，using the contents of NO ，IL-6 and TNF-α as indexes，uniform design was used to determine the optimal ratio of each ingredient in CGIF. Totally 240 mice were taken and randomly divided into 4 batches，with 60 mice in each batch. Each batch of mice was randomly divided into blank group （normal saline ），model group （normal saline ），positive drug group [levamisole ，4 mg/（kg·d）]，and optimal proportion extract of CGIF low-dose ，medium-dose and high-dose groups [ 0.952 8，1.905 6，3.811 2 g/（kg·d）]，with 10 mice in each group ；they were given medicine intragastrically ，qd，for consecutive 30 d. Except for blank group，mice in the other groups were intraperitoneally injected with cyclophosphamide [ 40 mg/（kg·d）] on the 24th day after first administration，qd，for consecutive 3 d to induce immunocompromised model. The immune activity of the optimal proportion extract was evaluated by determining visceral coefficients ，spleen lymphocyte transformation capacity ，serum contents of hemolysin，IL-2，IgM，IgG and IgA ，phagocytosis function of peritoneal macrophages. Another 20 mice were collected and given the optimal proportion extract 20 mL/kg intragastrically ，twice；acute toxicity of the formula was investigated with oral maximum tolerated dose （MTD）. RESULTS ：The optimal ratio of CGIF was that crude drug mass ratio of P. ginseng ， membranaceus，P. sibiricum ，L. chinensis was 1 ∶ 2 ∶ 2 ∶ 4. The immunological activity experiment showed that theoptimal proportion extract can significantly improve visceral indexes of mice ， spleen lymphocyte proliferation ability serum contents of hemolysin ，IL-2，IgM，IgG and IgA as well as macrophage phagocy tosis ability （P＜0.05 or P＜ 0.01）. The acute toxicity test indicated that oral MTD was over 15 g/kg，which was non-toxic. CONCLUSIONS ：The optimal proportion extract of CGIF can significantly enhance the immune function of mice and are non-toxic.

OBJECTIVE To analyze the changes of volatile co mponents in Olibanum and its processed products ，and to determine the contents of 4 components as octyl acetate. METHODS The volatile oil of Olibanum ，fried Olibanum and Olibanum stir-baked with vinegar were extracted. The components of volatile components were identified by GC-MS. The structure identification and data analysis of the chemical components with similarity ≥80％ were performed by using Xcalibur 4.0 software and NIST 2.0 mass spectrum database. The peak area normalization method was used to calculate the relative content of each component. GC method was adopted to simultaneously determine and compare the contents of limonene ，octyl acetate ，linalool and n-octanol in volatile components of Olibanum and its processed products. RESULTS Thirteen components were identified from volatile components of Olibanum ，fried Olibanum and Olibanum stir-baked with vinegar ，mainly including alcohols ，olefins and esters；among them ，relative contents of octyl acetate in Olibanum ，fried Olibanum and Olibanum stir-baked with vinegar were higher，which were 23.86％ ，37.80％ and 53.86％ respectively. The linear ranges of limonene ，octyl acetate ，linalool and n-octanol were 0.006 6-0.066 4，0.179 2 -1.792 0，0.003 7-0.037 0 and 0.032 8-0.328 0（r＞0.999 5）respectively；RSDs of precision，repeatability and stability （24 h）tests were all less than 2％；average recoveries were 98.56％，100.02％，99.13％ and 98.66％，respectively（RSD≤2.16，n＝6）. Average contents of 4 components in Olibanum were 0.15％，16.27％，0.36％ and 2.26％，while those of fried Olibanum were 0.85％，17.58％，0.66％ and 3.47％，respectively；those of Olibanum stir-baked with vinegar were 0.50％，19.75％，0.58％ and 3.34％，respectively. Compared with Olibanum ，average contents of octyl acetate ， linalool，n-octanol and limonene in volatile components of fried Olibanum and Olibanum stir-baked with vinegar were increased significantly（P＜0.05 or P＜0.01）. Compared with fried Olibanum ，average contents of limonene ，linalool and n-octanol were decreased significantly ，while those of octyl acetate were increased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS After fried and stir-baked with vinegar ，the volatile components in Olibanum are similar ，but the relative contents are different ，and the contents of octyl acetate and other components are increased.

Bacterial Infections ; diagnosis ; urine ; Colorimetry ; Female ; Humans ; Male ; Middle Aged ; Peritonitis ; diagnosis ; urine ; Urinalysis ; methods