OBJECTIVE To observe the clinical efficacy of Maitong Jun'an Decoction in treating coronary heart disease(CHD)angina of qi deficiency and blood stasis type,and preliminarily elucidate its possible mechanism of action through transcriptomics meth-ods.METHODS A total of 140 patients with CHD angina of qi deficiency and blood stasis type were included and randomly divided into a treatment group and a control group,with 70 cases in each group.During the treatment period,3 patients in the control group dropped out.The control group received basic Western medicine treatment for secondary prevention of CHD,while the treatment group received Maitong Jun'an Decoction in addition to the treatment in the control group.The treatment period for both groups was 8 weeks.Before and after treatment,the patients in both groups were evaluated for the TCM syndrome score,Canadian Cardiovascular Society(CCS)angina grading,Seattle angina questionnaire(SAQ)score,self-rating anxiety scale(SAS),self-rating depression scale(SDS)score,and adverse reactions.The peripheral blood of 9 patients before and after treatment was selected for transcriptomic sequencing based on the principle of gender,age,and disease duration matching.RESULTS After treatment,the TCM syndrome scores and total scores of the 2 groups were significantly reduced(P<0.01).The treatment group was better than the control group in improving chest pain,chest tightness,shortness of breath,fatigue and total score(P<0.05,P<0.01);the overall improvement rate of CCS angina grading in the treatment group was better than that in the control group(P<0.05);the SAQ,SAS and SDS scores of the 2 groups were significantly reduced before and after treatment(P<0.01),and the SAQ score of the treatment group was improved better than that of the control group(P<0.05,P<0.01).The transcriptomics results showed that there were 862 significantly different mR-NAs before and after treatment,including 509 up-regulated and 353 down-regulated.GO analysis showed that there were 666 biologi-cal processes in the differentially expressed mRNAs,mainly including viral gene expression,translation initiation,RNA catabolism,etc.There were 112 cell components,mainly including focal adhesion,ribosome subunit,nuclear spot,etc.There were 94 molecular functions,mainly including double-stranded RNA binding,cadherin binding,transcription co-regulatory factor activity,etc.KEGG analysis showed that the differentially expressed mRNAs enriched in 20 signaling pathways,mainly including glycerophospholipid me-tabolism pathway,AMPK signaling pathway,ribosome pathway,etc.CONCLUSION Maitong Jun'an Decoction can improve clini-cal symptoms in patients with CHD angina of qi deficiency and blood stasis type.Its mechanism of action is multi-target and multi pathway,mainly related to the regulation of glycerophospholipid metabolism pathway,AMPK signaling pathway,ribosome pathway.

OBJECTIVE To investigate and analyze the operational costs and current charging policies of pharmacy intravenous admixture service （PIVAS） in China， and provide a reference for promoting high-quality and sustainable development of PIVAS. METHODS Questionnaires were distributed in 30 provinces， autonomous regions， and municipalities across the country through the “Wenjuanxing” platform from May 6th to July 1st， 2022. The operational costs， charging status and suggestions of PIVAS were investigated and analyzed. RESULTS A total of 761 PIVAS participated in the survey nationwide， including 666 tertiary medical institutions， 93 secondary medical institutions， and 2 primary medical institutions. Approximately 60.58% of PIVAS had implemented a charging system that allowed charges. Among them， most PIVAS required inspection and evaluation before charging. The annual operating cost of PIVAS in China was approximately 2 098 100 yuan， with the integrated operating cost comprising 89.36% of the total， while the dispensing cost accounted for only 10.64%. Human costs emerged as the highest annual consumption （74.20%）， followed by decoration and facility maintenance costs （4.77%） and equipment acquisition costs （3.44%）. Regarding charges for different drugs nationwide， common drugs had an average charge standard of 4.39 yuan per bag while antibacterial drugs averaged 5.01 yuan per bag； hazardous drugs had an average charge of 23.17 yuan per bag， whereas parenteral nutrition solutions averaged 38.75 yuan per bag. However， the recommended average charges of the four drugs mentioned above were 6.71， 9.63， 38.35 and 44.03 yuan per bag， respectively. CONCLUSIONS At present， there is no unified inspection and evaluation standard and charging standard in China. Moreover， the current charging standard is lower than the recommended standard. It is necessary to combine operational costs and develop more reasonable and fair charging standards.

OBJECTIVE To explore the effects and mechanisms of cycloartenol on myocardial ischemia-reperfusion injury in mice.METHODS In vitro experiments,primary cardiomyocytes were extracted from 1-3 days SD mice.The hypoxia/reoxygenation model was established by incubating cells in a hypoxic culture box for 3 hours followed by reoxygenation in a normal culture box for 3 hours.The primary cardiomyocytes were divided into Control group,H/R group,low-dose(3 μmol·L-1)and high-dose(10 μmol·L-1)cycloartenol groups,and SB203580 group.CCK-8 was used to detect cell viability,the apoptosis rate was detected by flow cytometry,and Western blot was used to detect the expression levels of p38 MAPK and p-p38 MAPK in each group.In an in vi-vo experiment,7-week-old male C57BL/6J mice were randomly divided into a Control group,an I/R group,and three doses of cyclo-artenol(0.2,0.5,1.0 mg·kg-1)groups.The mice were continuously administered for seven days before the surgery.The model was prepared by ligation of the left anterior descending coronary artery(LAD)for 30 minutes,followed by reperfusion for 24 hours to in-duce myocardial ischemia-reperfusion injury.Left ventricular ejection fraction(LVEF),cardiac output(CO),left ventricular fractional shortening(LVFS)of each group of mice were detected by small animal ultrasound.TTC staining was used to detect the changes of is-chemic infarct size in each group.The changes of myocardial tissue in each group were observed by HE staining.The expression levels of p38 MAPK,p-p38 MAPK,IL-6 and TNF-α in myocardial tissue of mice were detected by Western blot.Serum levels of creatine ki-nase isoenzyme(CK-MB),lactate dehydrogenase(LDH),cardiac troponin I(cTnI),interleukin-6(IL-6),and tumor necrosis fac-tor-α(TNF-α)were measured using ELISA kits.RESULTS In vitro experiments demonstrated that compared with the H/R group,both the cycloartenol and SB203580 pretreatment groups showed a significant increase in myocardial cell viability and the apoptosis rate decrease,which can downregulate the protein expression level of p-p38 MAPK and decrease the ratio of p-p38 MAPK/p38 MAPK(P<0.05).In vivo experiments confirmed that compared with the I/R group,cycloartenol pretreatment significantly improved LVEF,LVFS,and CO values(P<0.05),reduce myocardial ischemic infarct size,thereby enhancing myocardial function.The protein ex-pression level of p-p38 MAPK in myocardial tissue was down-regulated,the ratio of p-p38 MAPK/p38 MAPK was decreased,and the expression levels of IL-6 and TNF-α were decreased.Additionally,cycloartenol pretreatment reduced the levels of CK-MB,LDH,cT-nI,IL-6,and TNF-α in mouse serum(P<0.05).CONCLUSION Pre-treatment with cycloartenol can protect mouse cardiac func-tion and alleviate myocardial ischemia-reperfusion injury.Its mechanism of action may be related to the inhibition of p38 MAPK phos-phorylation,reducing inflammatory reactions.

Objective·To investigate the effects of sennoside A(SA)on the formation of atherosclerotic plaque and the expression of 5-hydroxytryptamine(5-HT)and its receptor in mice with diabetes mellitus type 2(T2DM).Methods·Twelve mice with knocked-out apolipoprotein E gene were randomly divided into two groups,namely the model group and the model+SA group,with six mice in each group.Six C57BL/6J mice with the same genetic background were used as the control group.The control group was fed with normal diet,and the model group and the model+SA group were given intraperitoneal injection of streptozotocin(30 mg/kg)daily on the basis of high-fat diet to establish a model of T2DM.The model+SA group was given SA daily by gavage for 8 weeks,and the control group and the model group were given equal volume of distillation-distillation H2O by gavage.The body weight,fasting blood glucose(FBG)and 2-h postprandial blood glucose of mice were compared before and after modeling and treatment.The area of aortic plaque was observed by oil red O staining and hematoxylin-eosin(H-E)staining,and the level of 5-HT in serum and thoracic aorta was measured by ELISA kit.Western blotting was used to detect the expression of 5-hydroxytryptamine receptor 2B(HTR2B)and serotonin transporter(SERT)in thoracic aorta of mice.Results·Compared with the control group,the body weight,FBG and 2-h postprandial blood glucose in the model group increased,and glucose metabolism was disordered.The expression of HTR2B and SERT protein in thoracic aorta increased,while the concentration of 5-HT in thoracic aorta decreased.The serum 5-HT concentration increased(all P<0.05).After treatment with SA,compared with the model group,the body weight of the model+SA group decreased,and FBG and 2-h postprandial blood glucose were significantly improved.The area of aortic plaque and the expression of HTR2B and SERT protein in thoracic aorta significantly decreased,while the concentration of 5-HT increased.The serum 5-HT concentration decreased(all P<0.05).Conclusion·SA can reduce atherosclerotic plaque area in T2DM mice,which may be related to lowering blood glucose and inhibiting the expression of 5-HT and its receptor.

Background: s/Aims: Transmembrane 6 superfamily member 2 (TM6SF2) E167K variant is closely associated with the occurrence and development of metabolic dysfunction-associated steatotic liver disease (MASLD). However, the role and mechanism of TM6SF2 E167K variant during MASLD progression are not yet fully understood. Methods: The Tm6sf2167K knock-in (KI) mice were subjected to high-fat diet (HFD). Hepatic lipid levels of Tm6sf2167K KI mice were detected by lipidomics analysis. Thin-layer chromatography (TLC) was used to measure the newly synthesized triglyceride (TG) and phosphatidylcholine (PC). Results: The TM6SF2 E167K variant significantly aggravated hepatic steatosis and injury in HFD-induced mice. Decreased polyunsaturated PC level and increased polyunsaturated TG level were found in liver tissue of HFDinduced Tm6sf2167K KI mice. Mechanistic studies demonstrated that the TM6SF2 E167K variant increased the interaction between TM6SF2 and PNPLA3, and impaired PNPLA3-mediated transfer of polyunsaturated fatty acids (PUFAs) from TG to PC. The TM6SF2 E167K variant increased the level of fatty acid-induced malondialdehyde and reactive oxygen species, and decreased fatty acid-downregulated cell membrane fluidity. Additionally, the TM6SF2 E167K variant decreased the level of hepatic PC containing C18:3, and dietary supplementation of PC containing C18:3 significantly attenuated the TM6SF2 E167K-induced hepatic steatosis and injury in HFD-fed mice. Conclusions The TM6SF2 E167K variant could promote its interaction with PNPLA3 and inhibit PNPLA3-mediated transfer of PUFAs from TG to PC, resulting in the hepatic steatosis and injury during MASLD progression. PC containing C18:3 could act as a potential therapeutic supplement for MASLD patients carrying the TM6SF2 E167K variant.

Objective To investigate the application value of tenofovir alafenamide fumarate (TAF) in elderly patients with chronic hepatitis B (CHB) and its influence on bones and kidneys. Methods A total of 36 CHB patients, aged ≥60 years, who received TAF antiviral therapy in Qingdao Municipal Hospital, The Affiliated Hospital of Qingdao University, Qingdao Sixth People's Hospital, Chengyang People's Hospital, and Jimo People's Hospital from June 2021 to October 2022 were enrolled in this study, and all patients received TAF (25 mg/d) antiviral therapy. Related data were collected at baseline and weeks 24 and 48 of treatment, including virological indicators, biochemical parameters, urinary protein electrophoresis indices, transient elastography (FibroScan), and bone mineral density. Virological indicators included high-sensitivity HBV DNA quantification; biochemical parameters included total bilirubin, direct bilirubin (DBil), indirect bilirubin (IBil), alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, gamma-glutamyl transpeptidase, total bile acid (TBA), glucose, blood urea nitrogen, creatinine, estimated glomerular filtration rate, and cystatin C (Cys C); urinary protein electrophoresis indices included urinary β2 microglobulin (β2-MG), urinary retinol (URBP), and urinary α1 microspherin (α1-MG). The paired t -test was used for comparison of normally distributed continuous data before and after treatment, and the Wilcoxon signed-rank test was used for comparison of non-normally distributed continuous data before and after treatment; the chi-square test or the Fisher's exact test was used for comparison of categorical data. Results A total of 36 CHB patients completed 24 weeks of follow-up. The complete virological response rate after 24 weeks of treatment was higher than that at baseline [83.3% (30/36) vs 77.8% (28/36), χ 2 =0.36, P =0.55], and there were significant reductions in DBil ( t =-2.42, P =0.02) and Cys C ( t =-4.34, P < 0.001) from baseline to week 24. A total of 18 CHB patients completed 48 weeks of follow-up. The complete virological response rate after 48 weeks of treatment was higher than that at baseline (94.4% vs 77.8%, χ 2 =2.22, P =0.34), and there were significant increases in IBil ( t =2.43, P =0.03), TBA ( Z =-2.24, P =0.03), and bone mineral density T score of lumbar vertebra ( t =2.92, P = 0.01) and femoral neck ( t =2.42, P =0.03) and a significant reduction in liver stiffness measurement ( t =-2.31, P =0.03). There were no significant changes in β2-MG, URBP, and α1-MG after treatment (all P > 0.05). Conclusion TAF has a good antiviral effect in CHB patients aged ≥60 years and can help more CHB patients achieve complete virological response, without causing damage to the kidney, and it can also improve bone mineral density and liver fibrosis degree.

ObjectiveTo investigate the association of peroxisome proliferator-activated receptor gamma coactivator 1 alpha (PPARGC1A) rs8192678 single nucleotide polymorphism (SNP) with the risk of nonalcoholic fatty liver disease (NAFLD) and the influence of PPARGC1A rs8192678 SNP on NAFLD-related biochemical parameters. MethodsA total of 119 NAFLD patients who attended Qingdao Municipal Hospital Affiliated to Qingdao University from December 2017 to December 2018 were enrolled as NAFLD group, and 213 individuals who underwent physical examination during the same period of time were enrolled as control group. Clinical data and blood samples were collected from all subjects to measure related biochemical parameters and detect PPARGC1A rs8192678 SNP. The chi-square test was used to determine whether the genotype distribution of samples was in accordance with the Hardy-Weinberg equilibrium. The t-test or the Wilcoxon rank-sum test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. A binary logistic regression analysis was used to investigate the risk factors for NAFLD. ResultsThere were no significant differences in the genotype and allele frequencies of PPARGC1A rs8192678 between the NAFLD group and the control group (χ2=0.011 and 0.015, P=0.918 and 0.904). The binary logistic regression analysis showed that CT genotype of PPARGC1A rs8192678 was not a risk factor for NAFLD (odds ratio=0.951, 95% confidence interval: 0.368-2.457, P=0.918). In the NAFLD group, the patients carrying CT genotype had a significantly higher level of gamma-glutamyl transpeptidase (GGT) than those carrying CC genotype (Z=-2.331, P=0.020). ConclusionPPARGC1A rs8192678 SNP does not increase the risk of NAFLD, while NAFLD patients carrying CT genotype tend to have a higher serum level of GGT.

Objective To study the epidemiological characteristics of norovirus (NV) and rotavirus (RV) in children with diarrhea. Methods In this study, fecal specimens of 2 135 children with diarrhea admitted to our hospital from January 2019 to December 2019 were collected. The infection of NV and RV virus was detected by immunochromatography and ELISA, respectively. The epidemiological characteristics of RV virus. Results The total positive detection rate of 1 388 children was 65.01% (1 388/2 135), and the positive detection rate of RV virus was 34.52% (651/2 135), which was higher than 30.49% (737/2 135) of NV virus (χ²=7.895, P<0.05); there is no difference in the positive detection rate of NV virus among children of different genders (P>0.05), the positive detection rate of RV in males is 61.74%, higher than that of females 38.26% (χ²=25.318 , P<0.001); the age of children infected with NV and RV viruses are within 2 years old, and the cumulative positive detection rates are 64.21% (418/651) and 60.92% (449/737), which is higher than other ages (χ²=35.791, P<0.001); the high incidence season of NV virus is mainly winter, and the cumulative positive detection rate from October to December is 81.26% (529/651), higher than other seasons (χ²=173.426 , P<0.001), the high incidence season of RV virus is mainly spring and winter, the cumulative positive detection rate from November to March of the next year is 87.11% (642/737), higher than other seasons (χ²=252.628, P<0.001). Conclusion Spring and winter are the seasons of high incidence of viral diarrhea in children. Close monitoring of children under 2 years of age should be strengthened to prevent viral diarrhea early.

Objective:To construct a transmembrane 6 superfamily member 2 (Tm6sf2) E167K gene knock-in mouse model.Methods:The plasmid was constructed to simultaneously express the single-stranded guide RNA Cas9 at a specific site of the mouse Tm6sf2 gene in the donor plasmid carrying the Tm6sf2 E167K fragment. The above two plasmids were injected into the mouse fertilized eggs together. The positive F0 generation mice were validated by PCR detection and sequencing. The number of F2 generation surviving mice in three genotypes of wild (Wt), heterozygous and knock-in (KI) were calculated. Wt and KI male mice (8 mice/ group) of F2 generation littermates were selected and given a normal diet for 8 weeks. The body weight of the mice was recorded every week, and the glucose metabolism and lipid metabolism indexes of the two mice were detected. The comparison between groups was performed with an independent sample t-test.Results:Genotype detection and sequencing results showed that the Tm6sf2 E167K gene knock-in mouse model was successfully established. KI mice had absence of homozygous lethal embryo phenotype. The body weight of KI mice was higher than that of Wt mice during lactation, and the difference between the two groups was statistically significant ( P < 0.05).The fasting blood glucose of KI mice (9.50 ± 0.33)mmol/L was higher than that of Wt mice (7.80 ± 0.30)mmol/L, and the difference between the two groups was statistically significant ( P < 0.05). During the oral glucose tolerance test, the 2-hour blood glucose level of KI mice (9.20 ± 0.51)mmol/L was higher than that of Wt mice (7.60 ± 0.18)mmol/L, and the difference between the two groups was statistically significant ( P < 0.05). The liver triglyceride content of KI mice (8.40 ± 0.55)mg/g was higher than that of Wt mice (7.30 ± 0.63)mg/g, but the difference was not statistically significant ( P > 0.05). There was no significant difference in plasma triglyceride levels between the two mice ( P > 0.05). The Oil red O staining results showed that KI mice had more lipid accumulation in the centrilobular region of ??liver than Wt mice. Conclusion:Tm6sf2 E167K gene knock-in mice were successfully constructed. Tm6sf2 E167K gene knock-in can cause abnormal glucose metabolism in mice and promote the occurrence of hepatic steatosis.

Sennoside A (SA) is a bioactive component of Chinese herbal medicines with an activity of irritant laxative, which is often used in the treatment of constipation and obesity. However, its activity remains unknown in the regulation of insulin sensitivity. In this study, the impact of SA on insulin sensitivity was tested in high fat diet (HFD)-induced obese mice through dietary supplementation. At a dosage of 30 mg/kg/day, SA improved insulin sensitivity in the mice after 8-week treatment as indicated by HOMA-IR (homeostatic model assessment for insulin resistance) and glucose tolerance test (GTT). SA restored plasma level of glucagon-like peptide 1 (GLP1) by 90% and mRNA expression of by 80% in the large intestine of HFD mice. In the mechanism, SA restored the gut microbiota profile, short chain fatty acids (SCFAs), and mucosal structure in the colon. A mitochondrial stress was observed in the enterocytes of HFD mice with ATP elevation, structural damage, and complex dysfunction. The mitochondrial response was induced in enterocytes by the dietary fat as the same responses were induced by palmitic acid in the cell culture. The mitochondrial response was inhibited in HFD mice by SA treatment. These data suggest that SA may restore the function of microbiota-GLP1 axis to improve glucose metabolism in the obese mice.