ObjectiveTo establish the specific chromatogram of Chuanxiong Rhizoma dispensing granules（CRdg）， and to evaluate its quality by chemometrics and two reference substances for determination of multiple components（TRSDMC）. MethodsHigh performance liquid chromatography（HPLC） specific chromatograms were established using 13 batches of CRdg from 7 manufacturers， and preliminary quality evaluation was performed by similarity evaluation and chemometrics analysis. Eight characteristic peaks in the specific chromatogram of CRdg were measured on 22 different types of C₁₈ columns， and the actual retention times were recorded. Taking chlorogenic acid（peak 1） and senkyunolide A（peak 8） as double standard compounds， the retention times of the eight characteristic peaks were predicted by linear calibration using two reference substances（LCTRS）， and the method was validated on three other columns of different brands. Taking chlorogenic acid as reference peak， the relative correction factor method（RCFM） was used to quantify cryptochlorogenic acid， caffeic acid， ferulic acid， senkyunolide I and senkyunolide A， and the results were compared with the external standard method（ESM）. ResultsThe similarities of specific chromatograms of 13 batches of CRdg were all >0.90， and a total of 8 characteristic peaks were calibrated， and six of them were identified， including chlorogenic acid（peak 1）， cryptochlorogenic acid（peak 2）， caffeic acid（peak 3）， ferulic acid（peak 5）， senkyunolide I（peak 6） and senkyunolide A（peak 8）. Through chemometric analysis， it was found that ferulic acid， chlorogenic acid， senkyunolide I and cryptochlorogenic acid were the main components causing quality difference in CRdg， and the accuracy of LCTRS in predicting the retention time of 8 characteristic peaks was superior to that of the relative retention time method（RRT）. Further comparison of the results obtained from RCFM and ESM showed that there was no statistically significant difference between the two methods. ConclusionA quality evaluation method for CRdg based on HPLC specific chromatogram and TRSDMC is established， its qualitative accuracy is better than that of RRT， the quantitative accuracy is similar to that of ESM， and 4 quality-differentiated components among different manufacturers are found. This method is stable and reliable， and has reference value for the quality evaluation of other dispensing granules.

ObjectiveTo establish the specific chromatogram of Chuanxiong Rhizoma dispensing granules（CRdg）， and to evaluate its quality by chemometrics and two reference substances for determination of multiple components（TRSDMC）. MethodsHigh performance liquid chromatography（HPLC） specific chromatograms were established using 13 batches of CRdg from 7 manufacturers， and preliminary quality evaluation was performed by similarity evaluation and chemometrics analysis. Eight characteristic peaks in the specific chromatogram of CRdg were measured on 22 different types of C₁₈ columns， and the actual retention times were recorded. Taking chlorogenic acid（peak 1） and senkyunolide A（peak 8） as double standard compounds， the retention times of the eight characteristic peaks were predicted by linear calibration using two reference substances（LCTRS）， and the method was validated on three other columns of different brands. Taking chlorogenic acid as reference peak， the relative correction factor method（RCFM） was used to quantify cryptochlorogenic acid， caffeic acid， ferulic acid， senkyunolide I and senkyunolide A， and the results were compared with the external standard method（ESM）. ResultsThe similarities of specific chromatograms of 13 batches of CRdg were all >0.90， and a total of 8 characteristic peaks were calibrated， and six of them were identified， including chlorogenic acid（peak 1）， cryptochlorogenic acid（peak 2）， caffeic acid（peak 3）， ferulic acid（peak 5）， senkyunolide I（peak 6） and senkyunolide A（peak 8）. Through chemometric analysis， it was found that ferulic acid， chlorogenic acid， senkyunolide I and cryptochlorogenic acid were the main components causing quality difference in CRdg， and the accuracy of LCTRS in predicting the retention time of 8 characteristic peaks was superior to that of the relative retention time method（RRT）. Further comparison of the results obtained from RCFM and ESM showed that there was no statistically significant difference between the two methods. ConclusionA quality evaluation method for CRdg based on HPLC specific chromatogram and TRSDMC is established， its qualitative accuracy is better than that of RRT， the quantitative accuracy is similar to that of ESM， and 4 quality-differentiated components among different manufacturers are found. This method is stable and reliable， and has reference value for the quality evaluation of other dispensing granules.

ObjectiveTo establish the quality standards for Sennae Folium（Cassia angustifolia） dispensing granules based on standard decoctions. MethodsHigh performance liquid chromatography（HPLC） specific chromatograms were established for 15 batches of Sennae Folium（C. angustifolia） standard decoctions and 10 of Sennae Folium（C. angustifolia） dispensing granules from different manufacturers， and the similarity evaluation， hierarchical cluster analysis（HCA） and principal component analysis（PCA） were performed. Linear calibration with two reference substances（LCTRS） and quantitative analysis of multi-components by single-marker（QAMS） were established for the common peaks in the specific chromatograms to determine the contents of main components in the decoction pieces， standard decoctions and dispensing granules， and to calculate their transfer rates from decoction pieces to standard decoctions and dispensing granules. ResultsThe similarities of specific chromatograms of 15 batches of Sennae Folium（C. angustifolia） standard decoctions and 10 batches of Sennae Folium（C. angustifolia） dispensing granules were all greater than 0.95， and a total of 8 characteristic peaks were calibrated， and five of them were identified， including kaempferol-3，7-O-diglucoside， apigenin-6，8-di-C-glucoside， quercetin-3-O-gentianoside， sennoside B and sennoside A. HCA and PCA results showed that there were certain differences in the composition of different batches of standard decoctions， but no clustering was observed in the production area. As the standard decoctions， the extract rate of 15 batches of samples was 26.54%-45.38%， the contents of kaempferol-3，7-O-diglucoside， apigenin-6，8-di-C-glucoside， quercetin-3-O-gentianoside， sennoside B and sennoside A were 12.16-19.26， 2.57-4.94， 3.27-5.11， 6.75-11.39， 4.69-7.79 mg·g^-1， and their transfer rates from decoction pieces to standard decoctions were 45.41%-79.02%， 29.12%-55.07%， 40.52%-67.90%， 24.72%-49.12%， 27.54%-49.34%， respectively. The extract rates of Sennae Folium（C. angustifolia） dispensing granules（C8-C10） were 38.10%-39.50%， the transfer rates of the above five components from decoction pieces to dispensing granules were 72.85%-73.58%， 53.43%-53.94%， 40.19%-40.74%， 24.62%-25.00%， 28.65%-29.11%， respectively， which were generally consistent with the transfer rates from decoction pieces to standard decoctions. ConclusionCompared with the relative retention time method， LCTRS has higher prediction accuracy and is more suitable for chromatographic columns. The established quality control standard of Sennae Folium（C. angustifolia） dispensing granules based on standard decoction is reasonable and reliable， and all indicators of samples from different manufacturers are within the range specified based on the standard decoction， which can provide reference for the quality control and process research of this dispensing granules.

OBJECTIVE To evaluate the quality of Euscaphis japonica from different habitats. METHODS The relative correction factors of gallic acid， protocatechuic acid， ellagic acid， isoquercitrin， astragalin and apigenin were calculated with quercetin as the internal reference； the relative correction factors of euscaphic acid， oleanolic acid， stigmasterol and β-sitosterol were also calculated with ursolic acid as the internal reference. The contents of 12 components in 18 batches of samples were calculated by QAMS method and were compared with external standard method. At the same time， the contents of water-soluble extract， alcohol-soluble extract， total ash and acid-insoluble ash were detected. The quality of E. japonica was evaluated by principal component analysis （PCA）， orthogonal partial least squares-discriminant analysis （OPLS-DA）， and weighted technique for order preference by similarity to ideal solution （TOPSIS） method. RESULTS There was no significant difference between the results of QAMS method and external standard method for the 12 components in the 18 batches of samples. However， notable content variations were observed among different batches of samples. The results of PCA and OPLS-DA showed that S1-S7， S8- S12， and S13-S18 were clustered into one category respectively. Seven key characteristic components variable importance in projection values ＞1， euscaphic acid， ursolic acid， protocatechuic acid， apigenin， β-sitosterol， isoquercitrin， and oleanolic acid， respectively. The analysis results of the weighted TOPSIS method revealed that the relative closeness for evaluating the quality of 18 batches of samples ranged from 0.283 5 to 0.644 1， with the samples of E. japonica from Fengjie， Chongqing， demonstrating the highest quality. CONCLUSIONS The established method is accurate and feasible， which can be used for the quality evaluation of E. japonica combined with chemometrics and weighted TOPSIS model.

Aim To establish a reliable in vitro evaluation sys-tem to assess human liver UGTs activities and the inhibitory effect of drugs on human liver UGTs and validate its accuracy and stability.Methods Six probe substrate of the six major subtypes of human liver microsomal UGT enzymes were select-ed.The activity of human liver microsomal UGT enzymes was determined under optimized incubation conditions using a metab-olite generation method.The concentrations of metabolites were determined using a validated liquid chromatography-tandem mass spectrometry(LC-MS/MS)method.Known inhibitors were used to verify the established incubation system.Results The estab-lished LC-MS/MS analysis method complied with the require-ments of bioanalysis.In the incubation system of this experi-ment,known UGT inhibitors exhibited significant inhibitory effects.Conclusions This study successfully establishes an ac-curate and reliable in vitro evaluation system,which can be used to assess the in vitro inhibition of drugs on UGT enzymes,provi-ding important guidance for drug development and rational clini-cal medication.

The effect of porcine SIRT5 on replication of foot and mouth disease virus type O(FMDV-O)and the underlying regulatory mechanism were investigated.Western blot and RT-qPCR analyses were employed to monitor expression of endoge-nous SIRT5 in PK-15 cells infected with FMDV-O.Three pairs of SIRT5-specific siRNAs were synthesized.Changes to SIRT5 and FMDV-O protein and transcript levels,in addition to virus copy numbers,were measured by western blot and RT-qPCR analyses.PK-15 cells were transfected with a eukaryotic SIRT5 expression plasmid.Western blot and RT-qPCR analyses were used to explore the impact of SIRT5 overexpression on FMDV-O replication.Meanwhile,RT-qPCR analysis was used to detect the effect of SIRT5 overexpression on the mRNA expression levels of type I interferon-stimulated genes induced by SeV and FMDV-O.The results showed that expression of SIRT5 was up-regulated in PK-15 cells infected with FMDV-O and siRNA interfered with SIRT5 to inhibit FMDV-O replication.SIRT5 overexpression promoted FMDV-O replication.SIRT5 over-expression decreased mRNA expression levels of interferon-stimulated genes induced by SeV and FMDV-O.These results suggest that FMDV-O infection stimulated expression of SIRT5 in PK-15 cells,while SIRT5 promoted FMDV-O rep-lication by inhibiting production of type I interferon-stimula-ted genes.These findings provide a reference to further ex-plore the mechanism underlying the ability of porcine SIRT5 to promote FMDV-O replication.

Objective:To summarize the clinical, imaging, and pathological characteristics of mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes syndrome (MELAS) to improve the diagnosis of this rare disease.Methods:A retrospective case series was conducted to collect the clinical data and results of genetic testing, muscle biopsy, and imaging studies including computed tomography (CT), magnetic resonance imaging (MRI), and magnetic resonance spectroscopy (MRS) of 35 patients with MELAS admitted to the Nanjing Drum Tower Hospital from 2012 to 2021. Descriptive statistical analysis including mean, standard deviation, and frequency percentage were carried out.Results:The average age of onset of the patients was 30.2±2.3 years; the prevalence of family history was 20%. The two main initial symptoms were limb weakness and convulsions. The clinical manifestations of the neuromuscular system were proximal muscle weakness and exercise intolerance. The endocrine system is the most affected outside the neuromuscular system, with diabetes being the most common condition. Among the five patients who underwent brain CT, four showed hypodense lesions and two had calcified lesions. Brain MRI in 26 patients showed that the lesions more often affected the parietal lobe, basal ganglia, temporal lobe, occipital lobe, and frontal lobe than the infratentorial areas. Twelve of these individuals exhibited different levels of brain atrophy. Among the 10 patients who underwent 1H-MRS, nine showed a decrease in N-acetylaspartate (NAA) levels, eight exhibited abnormal lactate elevation (Lac peaks), whereas six had both reduced NAA levels and the presence of Lac peaks. Thirty-one patients underwent genetic testing; among them, 25 were found to have the mt.3243A>G mutation, while the remaining six exhibited rare gene alterations. Muscle biopsies were performed in 21 patients, and 15 showed abnormal mitochondrial proliferation manifested by ragged red fibers and defective oxidative phosphorylation manifested by cytochrome C oxidase (COX) enzyme-deficient muscle fibers. Conclusion:The clinical manifestations of MELAS syndrome are variable and complex, and early atypical symptoms could be missed or misdiagnosed. A detailed clinical history, imaging MRS analysis, muscle biopsy, and genetic testing are necessary to confirm the accurate diagnosis of MELAS.

Objective:To investigate the impact of ionizing radiation(IR)on the structure and function of the testis and pro-vide some strategies for the prevention and treatment of IR-induced damage(IRD).Methods:Using radiation dose simulation,se-men analysis,hormone testing,electron microscopy and single-cell transcriptome sequencing,we assessed and analyzed a case of IRD.We established a mouse model of IRD to validate the results of single-cell sequencing,and investigated the specific biological mecha-nisms of IRD and potential strategies for its intervention.Results:IR at 1-2 Gy significantly reduced sperm concentration and mo-tility,which gradually recovered after 12 months but the percentage of morphologically normal sperm remained low.It also caused im-balanced levels of various steroid hormones,decreased testosterone and dehydroepiandrosterone sulfate,increased progesterone,prolac-tin,luteinizing hormone,and follicle-stimulating hormone.Electron microscopy revealed damages to the testis structure,including loss of germ cells,atrophy of the seminiferous tubules,nuclear membrane depression of the spermatocytes,mitochondrial atrophy and de-formation,and reduction of mitochondrial cristae.Single-cell sequencing indicated significant changes in the function of the Leydig cells and macrophages and disrupted lipid-related metabolic pathways after IRD.Administration of L-carnitine to the mouse model im-proved lipid metabolism disorders and partially alleviated IRD to the germ cells.Conclusion:Ionizing radiation can cause disorders of testicular spermatogenesis and sexual hormones and inhibit lipid metabolism pathways in Leydig cells and macrophages.Improving lipid metabolism can alleviate IRD to germ cells.

Objective:To investigate the clinical characteristics and treatment mode changes of chronic pancreatitis (CP).Methods:The retrospective and descriptive study was conducted. The clinical data of 805 patients with CP who were admitted to the First Affiliated Hospital of Xi′an Jiaotong University from January 2013 to December 2022 were collected. There were 575 males and 230 females, aged 52(range, 10-87)years. Observation indicators: (1)demographic characteristics; (2)distribution of admission departments, hospitalization and treatment status. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distri-bution were represented as M(range). Count data were expressed as absolute numbers and percentages. The chi-square goodness-of-fit test was used for consistent between expected frequency and empirical distribution. Results:(1)Demographic characteristics. Of the 805 patients, there were 435 cases and 370 cases diagnosed as CP according to the primary and secondary diagnostic criteria, respec-tively, and the age of initial presentation of 805 patients was 52(range, 10-87)years. Among male and female patients, there was 1 and 0 case in the age group of 0-10 years, there were 16 and 14 cases in 11-20 years, 45 and 26 cases in 21-30 years, 82 and 30 cases in 31-40 years, 122 and 39 cases in 41-50 years, 157 and 51 cases in 51-60 years, 119 and 46 cases in 61-70 years, 31 and 21 cases in 71-80 years, 2 and 3 cases in 81-90 years, respectively. Patients aged 0-14, 15-34, 35-64, and ≥65 years were 0, 6, 28, and 9 for 2013, 1, 21, 34, and 10 for 2014, 1, 8, 38, and 7 for 2015, 0, 7, 52, and 10 for 2016, 0, 11, 35 and 9 for 2017, 1, 15, 72 and 23 for 2018, 0, 9, 55 and 11 for 2019, 2, 19, 58 and 16 for 2020, 0,20, 79 and 18 for 2021, 0, 25, 73 and 22 for 2022, respectively. Of the 805 patients, cases from Shaanxi Province, Gansu Province and other regions were 702, 48 and 55, respectively. There are 802 Han and 3 Hui patients, respectively, and the married, unmarried, divorced, and widowed patients were 732, 64, 7, and 2, respectively. Cases with blood type information was 682, with the distribution of blood types as 26.10%(178/682) of type A, 34.46%(235/682)of type B, 9.97%(68/682) of type AB, 29.47%(201/682)of type O, showing no significant difference compared to the distribu-tion of blood types in population of Shaanxi Province in 2022 (28.43% of type A, 30.50% of type B, 9.83% of type AB, and 30.50% of type O) ( χ2=0.50, P>0.05). (2)Distribution of departments, hospita-lization, and treatment. Of the 805 patients, cases admitted to the Department of Hepatobiliary Surgery, Department of Gastroenterology and other departments (Department of Geriatric Surgery, Infectious Diseases and General Surgery) were 594, 121 and 90, respectively. Cases of the number of hospitalizations in the Department of Hepatobiliary Surgery, Department of Gastroenterology and other departments were 771, 121 and 94, respectively. One patient might have been admitted for multiple times. The duration of hospital of stay of 805 patients was 11(rang, 1-67)days. The duration of hospital stay of patients admitted to the Department of Hepatobiliary Surgery, Department of Gastroenterology and other departments were 16(range, 2-48)days, 11(range, 5-19)days and 24(range, 12-35)days for 2013, 18(range, 2-63)days, 10(range, 3-29)days and 14(range, 7-30)days for 2014, 9(range, 1-35)days, 11(range, 2-16)days and 10(range, 5-33)days for 2015, 10(range, 1-55)days, 9(range, 4-16)days and 16(range, 4-27)days for 2016, 9(range, 2-38)days, 10(range, 4-20)days and 11(range, 5-27)days for 2017, 12(range, 3-46)days, 11(range, 2-26)days and 13(range, 7-27)days for 2018, 11(range, 1-33)days, 9(range, 3-23)days and 14(range, 4-17)days for 2019, 11(range, 1-67)days, 7(range, 1-23)days, and 16(range, 4-27)days for 2020, 10(range, 1-35)days, 8(range, 1-32)days and 14(range, 1-29)days for 2021, 9(range, 1-42)days, 12(range, 3-17)days and 11(range, 1-22)days for 2022. Of the 805 patients, cases receiving treatment as surgical treatment, pancreas extracorporeal shock wave lithotripsy (P-ESWL), endoscopic treatment and other treatments were 258, 117, 194 and 236, respectively. A total of 260 surgeries were performed on 258 patients receiving surgical treatment, and there were 236 cases receiving surgical treatment as the first choice, and 22 cases undergoing surgery after failed of other treatments. The surgical procedures included pancreaticoduodenectomy in 79 cases, distal pancreatectomy (with or without splenectomy, Peustow procedure) in 35 cases, partial pancreatectomy with pancreatic duct incision and stone removal and pancreaticojejunostomy in 77 cases, simple pancreatic duct incision and pancreaticojejunostomy in 58 cases, internal drainage for pancreatic pseudocysts in 4 cases, and drainage for removal of pancreatic necrotic tissue in 7 cases. There were 58 patients with post-operative complications, including 9 cases of bleeding, 19 cases of infection, 2 cases of gastric paresis, 21 cases pancreatic fistula, and 7 cases of intestinal leakage. Cases with postoperative complications were recovered and discharged after symptomatic treatment. Of the 117 patients receiving P-ESWL, the times of treatment per patient was 1.3±0.7, and a total of 154 times of P-ESWL were performed. There were 89 patients receiving one time of P-ESWL, and 28 patients receiving ≥2 times of P-ESWL. Of the 194 patients receiving endoscopic treatment, the times of treatment per patient was 1.2±0.6, and a total of 238 times of endoscopic treatment were performed. There were 103 cases receiving stent placement, 43 cases receiving nasopancreatic/nasobiliary drainage, 7 cases receiving pancreatic duct balloon dilation, 71 cases receiving pancreatic stone removal, and 58 cases receiving duodenal papilla sphincterotomy. There were 168 patients receiving one time of endoscopic treatment, and 26 patients receiving ≥2 times of endoscopic treatment.There was no postoperative complica-tions reported. Conclusions:The proportion of males in CP patients is relatively high, and the proportion gradually increases with age of 51-60 years. There is no significant difference in the distribution of blood types among CP patients and the population of Shaanxi Province. The proportion of minimally invasive treatment (extracorporeal shock wave lithotripsy and endoscopic therapy) in CP treatment is on the rise, and the duration of hospital stay for patients is on the decline.

Objective To investigate the feasibility of constructing the risk index of Echinococcus infection based on the classification of echinococcosis lesions, so as to provide insights into the management of echinococcosis. Methods The imaging data of echinococcosis cases were collected from epidemiological surveys of echinococcosis in China from 2012 to 2016, and the detection of incident echinococcosis cases was captured from the annual echinococcosis prevention and control reports across provinces (autonomous regions) and Xinjiang Production and Construction Corps in China from 2017 to 2022. After echinococcosis lesions were classified, a risk index of Echinococcus infection was constructed based on the principle of discrete distribution marginal probability and multi-group classification data tests. The correlation between the risk index of Echinococcus infection and the detection of incident echinococcosis cases was evaluated in the provinces (autonomous regions and corps) from 2017 to 2022, and the correlations between the short and medium-term risk indices and between the medium and long-term risk indices of Echinococcus infection were examined using a univariate linear regression model. Results A total of 4 014 echinococcosis cases in China from 2012 to 2016 were included in this study. The short-, medium- and long-term risk indices of E. granulosus infection varied in echinococcosis-endemic provinces (autonomous regions and corps) of China (χ² = 4.12 to 708.65, all P values < 0.05), with high short- (0.058), medium- (0.137) and long-term risk indices (0.104) in Tibet Autonomous Region, and the short-, medium- and long-term risk indices of E. multilocularis infection varied in echinococcosis-endemic provinces (autonomous regions and corps) of China (χ² = 6.74 to 122.60, all P values < 0.05), with a high short-term risk index in Sichuan Province (0.016) and high medium- (0.009) and long-term risk indices in Qinghai Province (0.018). There were no significant correlations between the risk index of E. granulosus infection and the detection of incident cystic echinococcosis cases during the study period (t = −0.518 to 2.265, all P values > 0.05), and strong correlations were found between the risk indices of E. multilocularis infection and the detection of incident alveolar echinococcosis cases (including mixed type) in 2018, 2020, 2021, 2022, during the period from 2017 through 2020, from 2017 through 2021, from 2017 through 2022 (all r values > 0.7, t = 2.521 to 3.692, all P values < 0.05). Linear regression models were established between the risk index of E. multilocular infection and the detection of alveolar echinococcosis cases (including mixed type), and the models were all statistically significant (b = 0.214 to 2.168, t = 2.458 to 3.692, F = 6.044 to 13.629, all P values < 0.05). The regression coefficients for the correlations between the medium- and short-term, and between the long- and medium-term risk indices of E. granulosus infection were 2.339 and 0.765, and the regression coefficients for the correlations between the medium- and short-term, and between the long- and medium-term risk indices of E. multilocular infection were 0.280 and 1.842, with statistical significance seen in both the regression coefficients and regression models (t = 16.479 to 197.304, F = 271.570 to 38 928.860, all P values < 0.05). Conclusions The risk index of Echinococcus infection has been successfully established based on the classification of echinococcosis lesions, which may provide insights into the prevention and control, prediction, diagnosis and treatment, and classified management of echinococcosis.