Objective To determine the pathological mechanism of gastrolithiasis formation in vitro and the effects of pectinase on gastrolithiasis. Methods The test in vitro was divided randomly into two groups:(1) Haws and persimmons(25g respectively) were chewed and put into two clean containers filled with fresh gastric juice.(2) Wine containing 30% alcohol(50ml) was added to the container. Each group included 4 collections.All two groups were placed in 37℃ calorstat box to observe the course and time of gastrolith formation. After gastrolith formation, 2g pectinase was added to the container and observed the decomposition of pectinase to gastrolith and the time was recorded. The effects of pectinase on 54 patients with gastrolithiasis were also investigated. Results The mean time of haws juice coagulation and persimmons was 23 minutes and 25 minutes respectively in the group without wine; but in the group with wine, the mean time of haws juice coagulation and persimmons was 14 minutes and 19 minutes respectively. After adding pectinase, the mean time of clot initiated decomposition was about 17 minutes in both groups, but complete decomposition needed 34 minutes. All the 54 patients with gastrolithiasis who received pectinase were cured.The gastrolith disappeared 6 hours after taking pectinase confirmed by barium meal examination. Conclusions Haws and/or persimmons mixed with gastric juice could induce gastrolith formation. The effects of pectinase on patients with haws or persimmons gastrolithiasis are good, this therapeutic procedure is simple, and the gastrolith resolves quickly.Pectinase may be widely used in the treatment of gastrolithiasis.

Objective To investigate the compatibility of Shuanghuanglian Injection mixed with Cefazolin Sodium for Injection, Dexamethasone Sodium Phosphate Injection and Ribavirin Injection. Methods Under room temperature, the changes of appearance were determined by pH meter and particle counter for Shuanghuanglian Injection mixed with Cefazolin Sodium for Injection, Dexamethasone Sodium Phosphate Injection and Ribavirin Injection. Results There were no evident changes in appearance and pH of Shuanghuanglian Injection mixed with Cefazolin Sodium for Injection, Dexamethasone Sodium Phosphate Injection and Ribavirin Injection. The number of particles (diameter≥25 μm) of Shuanghuanglian Injection mixed with Cefazolin Sodium for Injection and Dexamethasone Sodium Phosphate Injection was increased after 0.5 h. Conclusion Shuanghuanglian Injection can be mixed with Cefazolin Sodium for Injection, Dexamethasone Sodium Phosphate Injection and Ribavirin Injection for bigeminy, while should not for multitherapy.

The effect and mechanism of α-lipoic acid(ALA)on the injury of kidneys in diabetic rats induced by streptozotocin were investigated.Results showed that ALA decreased the level of oxidative stress,the production of advanced glycation end products(AGE)[(0.087±0.003 vs 0.103 4±0.014)pg/mg protein,P<0.05],and the expression of AGE receptor protein(1.8I±0.04 vs 2.67±0.01,P<0.01)and transforming growth factor-β(TGF-β)mRNA(1.51 4±0.20 vs 2.04±0.08,P<0.05)in renal cortex of diabetie rats,resulting in reduced kidney injury and improved renal function in diabetic rats.

As the most common organ-specific autoimmune disease,autoimmune thyroiditis (AIT) is characterized by intrathyroidal lymphocyte infiltration and specificthyroid autoantibodies.Modern medicine is short of effective etiological treatment at present,and has no specific medicine foreuthyroidism of AIT patients,who have been followed-up passively.Chinese medicine treatmenthas the characteristics of multiple-targets,multiple-links and multiple-pathways,which plays a special superiorityrole in the prevention and treatment of AIT.The purpose of this article was to sort out and set forthmore Chinese medicine pharmacology on AIT recently,which provided evidence for further clinical apply.

OBJECTIVETo evaluate the inhibitory effect of total bufadienolides from toad venom against H22 tumor in mice and preliminarily analyze the structures of the metabolites in tissues.

METHODHPLC and LC-MS were used for analysis of the chemical composition of TBFs. High, middle and low dosages of TBFs were orally administered or intra-peritoneally injected to H22 tumor-bearing mice for thirteen days. The animals were killed and the tumors were stripped and weighed. The metabolites in the tissues such as heart, liver, spleen, lung and kidney, were analyzed by HPLC and LC-MS.

RESULTThe chemical composition of TBFs were identified by comparison of the retention times with those of reference substances, on-line UV spectra and MS data. Its main components are concerned with gamabufotalin, arenobufagin, bufotalin, resibufagin, cinobufotalin, bufalin, cinobufagin and resibufogenin. TBFs had no obvious influence on body weight of H-22 tumor-bearing mice orally administered and the inhibition rate against tumor were 14.76%, 16.38% and 10.32% for low (5 mg x kg(-1)), middle (10 mg x kg(-1)) and high dosage (20 mg x kg(-1)), respectively. The mice intra-peritoneally injected with middle and high-dose of TBFs gained body weight slower than the control mice on the 5th day and recovered on the 13th day. The inhibition rate against tumor were 17.30%, 19.80% and 40.95% for low (1.5 mg x kg(-1)), middle (3 mg x kg(-1)) and high dose (6 mg x kg(-1)), respectively. The inhibitory effect took on dose-dependent manner. Based on the HPLC analyses on heart, liver, spleen, lung and kidney, bufadienolides were found in the liver tissue and 11 compounds of them were tentatively identified by LC-DAD-MS.

CONCLUSIONTBFs by oral administration had no inhibitory effect against H22 tumor in mice, however, TBFs by intra-peritoneal injection displayed the significantly inhibitory effect, accompanying some toxicity for early duration of the study. The identification of bufadienolides in the liver provides a good basis for the further investigation of the metabolic pathways of TBFs in vivo.

Amphibian Venoms ; chemistry ; Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; metabolism ; Bufanolides ; administration & dosage ; metabolism ; Carcinoma, Hepatocellular ; drug therapy ; metabolism ; physiopathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drug Administration Routes ; Female ; Humans ; Male ; Mice ; Mice, Inbred ICR ; Neoplasm Transplantation

OBJECTIVE:To establish the quality standard of Ligularia fischeri. METHODS:TLC was used for qualitative identification of samples. The contents of moisture,ash and extract were determined. The content of ferulic acid in samples was determined by HPLC. The determination was performed on Waters SunFire C18column with mobile phase consisted of acetonitrile-0.1% phosphoric acid(13:87,V/V)at the flow rate of 1.0 mL/min. The detection wavelength was set at 319 nm, and column temperature was 30 ℃. The sample size was 10 μL. RESULTS:TLC spots were clear and well-separated without interference from genitive control. The moisture,total ash,acid-insoluble ash and extract of samples were 7.6%-9.4%, 11.7%-19.6%,5.9%-14.1% and 25.4%-37.5%,respectively. The linear range of ferulic acid were 0.021 2-0.636 8 μg(r=0.999 9). limited of quantation was 2.25 ng,the limited of detection was 0.75 ng. RSDs of intermediate precision, reproducibility and stability tests were all lower than 3.0%. The recoveries ranged 97.81%-100.59%(RSD=1.02%,n=9). CONCLUSIONS:The moisture,total ash and acid-insoluble ash of samples are no more than 10.0%,19.0%,12.0%;the extract and the content of ferulic acid are no less than 25.0% and 0.1%. Established standard can provide reference for quality control of L. fischeri.

OBJECTIVE To establish the method for the content determination of ephedrine hydrochloride and pseudoephedrine hydrochloride in Shexiang zhuanggu plaster ，and to evaluate the quality of 222 batches of Shexiang zhuanggu plaster from 41 manufacturers. METHODS HPLC method was established. The determination was performed on Shimadzu Shim-pack GIS-C 18 column with mobile phase consisted of acetonitrile- 0.1％ phosphoric acid （3∶97，V/V）at the flow rate of 0.9 mL/min. The column temperature was set at 30 ℃，the detection wavelength was 210 nm，the sample size was 10 μL. Taking the content data of 222 batches of samples as index ，the cluster heatmap was drawn by Hiplot biomedical data visualization and analysis platform. RESULTS The results of the methodological investigation were in line with the requirements of the general principles stated in 2020 edition of Chinese Pharmacopoeia （part Ⅳ ）. The total contents of ephedrine hydrochloride and pseudoephedrine hydrochloride in 222 batches of samples from 41 manufacturers were 0.646-6.325 μg/cm2. Results of cluster heatmap analysis showed that these samples of 41 manufacturers could be divided into 3 categories. The contents of components in samples from different manufacturers varied greatly ，and the contents of components in different batches of samples from the same manufacturers also varied greatly. If the proposed limit of this product was that the total amount of ephedrine hydrochloride and pseudoephedrine hydrochloride per 1 cm2 would not be less than 1.00 μg，23 of the 222 batches of samples were unqualified. CONCLUSIONS The established method for the content determination of ephedrine hydrochloride and pseudoephedrine hydrochloride in Shexiang zhuanggu plaster is accurate and precise ，and can be used for the quality control. Some manufacturers should pay attention to optimizing production process and strengthening quality control.

OBJECTIVE：To optimize the primary processing technology of Gentiana rigesce ns. METHODS ：HPLC method was adopted for content determination of loganic acid ，swertiamarin and gentiopicroside in G. rigescens ，and overall desirability value （OD value ） of the contents of above 3 components was taken as index to carry out single factor test on blanching temperature，blanching time and drying temperature. Based on that ，Box-Behnken design-response surface methodology was used to optimize primary processing technology of G. rigescens . Validation test was also performed. The samples prepared by optimized technology were compared with those dried in the shade. RESULTS ：The optimal primary processing technology of G. rigescens included blanching time of 5 min，blanching temperature of 40 ℃ and drying temperature of 60 ℃. Validation test showed that the average OD value of the 3 components was 0.565 2，with a deviation of 0.94％ from the predicted value （0.570 6）. Compared with samples dried in the shade ，OD value of 3 components in samples prepared by optimized technology were increased significantly ， indicating the quality of the samples prepared by the optimized technology was better. CONCLUSIONS ：The optimal technology is stable and feasible ，and can be used for the primary processing of G. rigescens .

ObjectiveTo determine the mechanism of Yitangkang in correcting excessive apoptosis of skeletal muscle cells to improve insulin resistance （IR） by inhibiting the advanced glycation end product （AGE）/receptor for the advanced glycation end product （RAGE） signaling pathway. Method① In vitro experiments. Yitangkang-medicated serum was prepared. C2C12 cells were divided into a blank group， a model group， high-， medium-， and low-dose Yitangkang-medicated serum groups （40， 20， and 10 g·kg^-1）， and a RAGE inhibitor group. The IR model was induced by palmitic acid in C2C12 cells except for those in the blank group. After the corresponding intervention methods were conducted，the cell viability and glucose consumption level of each group were determined. In addition，the apoptosis rate was determined using flow cytometry. The mRNA and protein expression levels of the important apoptotic proteins ［B-cell lymphoma 2 （Bcl-2）， Bcl-2-associated X protein （Bax）， p53， cysteinyl aspartate-specific protease-3 （Caspase-3）， and cysteinyl aspartate-specific protease-9 （Caspase-9）］ were determined using Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. ② In vivo experiments. Ninety-six eligible Wistar rats were divided into a blank group， a model group， high-，medium-，and low-dose Yitangkang groups （40， 20， and 10 g·kg^-1）， and a western medicine group （pioglitazone hydrochloride，1.35 mg·kg^-1）. The IR model was induced using high-glucose and high-fat feed for diabetes combined with intraperitoneal injection of low-dose streptozotocin （STZ） in animals and verified by the hyperinsulinemic-euglycemic clamp （HEC） test. After the model was determined successfully， the rats in each group were given intragastric administration of drugs as required. Terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） assay was performed to determine the number of positive apoptotic cells in the skeletal muscle tissues of rats in each group，while Real-time polymerase chain reaction（Real-time PCR） and Western blot were performed to determine the mRNA and protein expression levels of the important apoptotic proteins Bcl-2， Bax， p53， Caspase-3， and Caspase-9. Result① In vitro experiments. compared with the blank group， the model groups showed increased apoptosis rate of C2C12 cells and decreased cell viability and glucose consumption （P<0.01）. Compared with the model group， the Yitangkang-medicated serum groups and the RAGE inhibitor group showed decreased apoptosis rate of C2C12 cells and increased cell viability and glucose consumption （P<0.01）. Compared with the blank group， the model group showed decreased expression levels of Bcl-2 mRNA and protein in C2C12 cells and increased mRNA and protein expression levels of Bax， p53， Caspase-3， and Caspase-9 （P<0.01）. Compared with the model group， the Yitangkang-medicated serum groups and the RAGE inhibitor group showed increased expression levels of Bcl-2 mRNA and protein in C2C12 cells （P<0.01） and decreased mRNA and protein expression levels of Bax， p53， Caspase-3， and Caspase-9 （P<0.05， P<0.01）. ② In vivo experiments. The number of positive apoptotic cells in the skeletal muscle tissues of rats in the model group significantly increased as compared with that in the blank group （P<0.01）. The number of positive apoptotic cells in the skeletal muscle tissues of rats in the Yitangkang groups and the western medicine group decreased as compared with that in the model group （P<0.01）. Compared with the blank group， the model group showed decreased expression levels of Bcl-2 mRNA and protein in skeletal muscle tissues of rats and increased mRNA and protein expression levels of Bax， p53， Caspase-3， and Caspase-9 （P<0.01）. Compared with the model group， the Yitangkang groups and the western medicine group showed increased expression levels of Bcl-2 mRNA and protein in skeletal muscle tissues of rats （P<0.01） and decreased mRNA and protein expression levels of Bax， p53， Caspase-3， and Caspase-9 （P<0.05， P<0.01）. The medium-dose Yitangkang showed a similar effect as RAGE inhibitor， and the effect was equivalent to that of pioglitazone hydrochloride. ConclusionYitangkang can inhibit skeletal muscle cell apoptosis by inhibiting the AGE/RAGE signaling pathway.