OBJECTIVE: Cassiae Semen (CS, Juemingzi in Chinese) is a widely used traditional Chinese medicine with a variety of pharmacological effects. This study aimed to investigate the potential therapeutic effects and molecular mechanisms of anthraquinones of CS (AQS) for adiposity. METHODS: The chemical components of the AQS were determined using high-performance liquid chromatography (HPLC). Network pharmacology analysis was used to predict potential anti-obesity targets of action for AQS. We constructed high fat with high sugar water diet-induced obese mice and observed their body weight and whole-body lipid metabolism to evaluate the efficacy of AQS in promoting lipid metabolism. Subsequently, the epidermal temperature at the brown adipose tissue (BAT) before and after cold stimulation was observed and the expression of lipid metabolism-related genes in the liver and BAT tissues was detected to clarify the mechanism of action of AQS. RESULTS: Network pharmacology analysis showed that AQS was involved in the regulation of liver and adipose tissue function under obesity. Pathological and biochemical results showed that AQS reduced lipid accumulation in the liver and adipose tissue induced by an unhealthy diet. With the increase of cold tolerance, the volume and weight of BAT were increased by AQS, suggesting that it regulated the body heat production dominated by BAT. After AQS treatment, the levels of genes related to uncoupling protein1 (UCP1)-mediated adaptive thermogenesis in BAT tissues and lipid metabolism in the liver were also increased, which further proved that AQS activated BAT function to promote lipid metabolism in the whole body. CONCLUSION This study revealed the pharmacological effects of AQS, thereby providing a scientific basis for regulating BAT thermogenesis and liver lipid metabolism to alleviate obesity and providing clues for further exploring the application of natural active ingredients in the treatment of metabolism-related diseases.

Objective:To study the anti-inflammatory and protective effects of electroacupuncture on rats with cardiopulmo-nary resuscitation based on cholinergic anti-inflammatory pathway(CAP).Methods:Sixty male SD rats were randomly divided into sham operation group(Sham group),cardiopulmonary resuscitation group,electroacupuncture group,antagonist of α7-nicotinic ace-tylcholine receptor(α7nAChR)α Bungarus toxin group(αBGT group,1 μg/kg)and agonist of α7nAChR 3-(2,4-dimethoxybenzyli-dene)anabaseine(GTS-21)group(5 mg/kg),with 12 rats in each group.Model of cardiac arrest was established by inducing ventric-ular fibrillation in rats with percutaneous epicardial electrical stimulation,and routine resuscitation was performed,the defibrillation times,cardiopulmonary resuscitation duration and 72 h survival rate of rats in each group were observed and recorded.Neurological damage of rats in each group was scored(NDS score).Levels of inflammatory factors TNF-α,IL-1β and IL-6 in serum,heart,lung and brain of rats were measured by ELISA.HE staining was applied to evaluate the degree of tissue damage of heart,lung and brain tis-sues of rats in each group.Expressions of α7nAChR,NF-κB/MAPKs signal pathway related proteins in heart,lung and brain of rats in each group were detected by Western blot.Results:Compared with Sham group,the number of defibrillations,duration of cardiopul-monary resuscitation,NDS score,levels of TNF-α,IL-1β and IL-6 in serum,heart,lung and brain tissues,and NF-κB p65 protein in heart,lung and brain tissues in cardiopulmonary resuscitation group increased greatly(P<0.05),the 72 h survival rate,phosphory-lation levels of α7nAChR protein,ERK1/2,JNK and p38MAPK in heart,lung and brain tissues decreased greatly after restoration of spontaneous circulation(P<0.05),cells and neurons in the CA1 region of heart,lung and hippocampus were severely damaged.Com-pared with cardiopulmonary resuscitation group,change trend of relevant indicators in electroacupuncture group was opposite to the above,damage of cells and neurons in rat heart,lung and brain tissue was reduced(P<0.05).αBGT reduced the anti-inflammatory effect of electroacupuncture on cardiopulmonary resuscitation rats(P<0.05).GTS-21 enhanced the anti-inflammatory protection of electroacupuncture on cardiopulmonary resuscitation rats(P<0.05).Conclusion:Electroacupuncture has anti-inflammatory and pro-tective effects on cardiopulmonary resuscitation rats based on CAP,and NF-κB/MAPKs participate in α7nAChR mediated CAP.

Objective To investigate the mechanism of modified Huanglian Wendan Decoction in improving insulin resistance(IR)HepG2 cells by regulating JAK2/STAT3 signaling pathway.Methods HepG2 cells were incubated with 0.25 mmol/L palmitic acid for 24 h to induce IR model.The cells were divided into model group,drug containing serum group and metformin hydrochloride group.Blank serum,modified Huanglian Wendan Decoction drug containing serum and 2 mmol/L metformin hydrochloride intervention were administered,respectively.Normal cultured HepG2 cells were used as control group.The glucose assay kit was used to detect the glucose content in the cell supernatant,the liver glycogen assay kit was used to detect the glycogen content in the cells,the triglycerides(TG)assay kit was used to detect the TG content in the cells,PAS staining was used to observe the glycogen status of the cells,oil red O staining was used to observe the lipid droplet status of the cells,ELISA assay kit was used to detect the contents of interleukin-6(IL-6)and tumor necrosis factor(TNF)-α in cell supernatant,Western blot was used to detect the protein expressions of Janus kinase 2(JAK2),signal transduction and transcription activator 3(STAT3),LC3 and Beclin-1 in the cells.Results Compared with the control group,the glucose content in the supernatant of HepG2 cells in the model group increased(P<0.01),the intracellular glycogen content decreased(P<0.01),the TG content increased(P<0.01),the glycogen staining area decreased(P<0.01),the lipid droplet staining area increased(P<0.01),the contents of IL-6 and TNF-α in the supernatant increased(P<0.01),the expressions of JAK2 and STAT3 proteins increased(P<0.01),and the protein expression of LC3Ⅱ/LC3Ⅰ and Beclin-1 decreased(P<0.01).Compared with the model group,the glucose content decreased in the drug containing serum group and metformin hydrochloride group(P<0.01),the intracellular glycogen content increased(P<0.01),the TG content decreased(P<0.01),the glycogen staining area increased(P<0.01),the lipid droplet staining area decreased(P<0.01),the contents of IL-6 and TNF-α in cell supernatant decreased(P<0.01),the expressions of JAK2 and STAT3 proteins decreased(P<0.01),and the expressions of LC3Ⅱ/LC3Ⅰ and Beclin-1 proteins increased(P<0.05,P<0.01).The effect of modified Huanglian Wendan Decoction was better than that of metformin hydrochloride(P<0.05,P<0.01).Conclusion Modified Huanglian Wendan Decoction can possibly improve glucose and lipid metabolism and reduce inflammation in IR-HepG2 cells through intervening JAK2/STAT3 signaling pathway and mediating autophagy.

Objective To investigate the mechanism of modified Huanglian Wendan Decoction in improving insulin resistance(IR)HepG2 cells by regulating JAK2/STAT3 signaling pathway.Methods HepG2 cells were incubated with 0.25 mmol/L palmitic acid for 24 h to induce IR model.The cells were divided into model group,drug containing serum group and metformin hydrochloride group.Blank serum,modified Huanglian Wendan Decoction drug containing serum and 2 mmol/L metformin hydrochloride intervention were administered,respectively.Normal cultured HepG2 cells were used as control group.The glucose assay kit was used to detect the glucose content in the cell supernatant,the liver glycogen assay kit was used to detect the glycogen content in the cells,the triglycerides(TG)assay kit was used to detect the TG content in the cells,PAS staining was used to observe the glycogen status of the cells,oil red O staining was used to observe the lipid droplet status of the cells,ELISA assay kit was used to detect the contents of interleukin-6(IL-6)and tumor necrosis factor(TNF)-α in cell supernatant,Western blot was used to detect the protein expressions of Janus kinase 2(JAK2),signal transduction and transcription activator 3(STAT3),LC3 and Beclin-1 in the cells.Results Compared with the control group,the glucose content in the supernatant of HepG2 cells in the model group increased(P<0.01),the intracellular glycogen content decreased(P<0.01),the TG content increased(P<0.01),the glycogen staining area decreased(P<0.01),the lipid droplet staining area increased(P<0.01),the contents of IL-6 and TNF-α in the supernatant increased(P<0.01),the expressions of JAK2 and STAT3 proteins increased(P<0.01),and the protein expression of LC3Ⅱ/LC3Ⅰ and Beclin-1 decreased(P<0.01).Compared with the model group,the glucose content decreased in the drug containing serum group and metformin hydrochloride group(P<0.01),the intracellular glycogen content increased(P<0.01),the TG content decreased(P<0.01),the glycogen staining area increased(P<0.01),the lipid droplet staining area decreased(P<0.01),the contents of IL-6 and TNF-α in cell supernatant decreased(P<0.01),the expressions of JAK2 and STAT3 proteins decreased(P<0.01),and the expressions of LC3Ⅱ/LC3Ⅰ and Beclin-1 proteins increased(P<0.05,P<0.01).The effect of modified Huanglian Wendan Decoction was better than that of metformin hydrochloride(P<0.05,P<0.01).Conclusion Modified Huanglian Wendan Decoction can possibly improve glucose and lipid metabolism and reduce inflammation in IR-HepG2 cells through intervening JAK2/STAT3 signaling pathway and mediating autophagy.

Objective:To study the anti-inflammatory and protective effects of electroacupuncture on rats with cardiopulmo-nary resuscitation based on cholinergic anti-inflammatory pathway(CAP).Methods:Sixty male SD rats were randomly divided into sham operation group(Sham group),cardiopulmonary resuscitation group,electroacupuncture group,antagonist of α7-nicotinic ace-tylcholine receptor(α7nAChR)α Bungarus toxin group(αBGT group,1 μg/kg)and agonist of α7nAChR 3-(2,4-dimethoxybenzyli-dene)anabaseine(GTS-21)group(5 mg/kg),with 12 rats in each group.Model of cardiac arrest was established by inducing ventric-ular fibrillation in rats with percutaneous epicardial electrical stimulation,and routine resuscitation was performed,the defibrillation times,cardiopulmonary resuscitation duration and 72 h survival rate of rats in each group were observed and recorded.Neurological damage of rats in each group was scored(NDS score).Levels of inflammatory factors TNF-α,IL-1β and IL-6 in serum,heart,lung and brain of rats were measured by ELISA.HE staining was applied to evaluate the degree of tissue damage of heart,lung and brain tis-sues of rats in each group.Expressions of α7nAChR,NF-κB/MAPKs signal pathway related proteins in heart,lung and brain of rats in each group were detected by Western blot.Results:Compared with Sham group,the number of defibrillations,duration of cardiopul-monary resuscitation,NDS score,levels of TNF-α,IL-1β and IL-6 in serum,heart,lung and brain tissues,and NF-κB p65 protein in heart,lung and brain tissues in cardiopulmonary resuscitation group increased greatly(P<0.05),the 72 h survival rate,phosphory-lation levels of α7nAChR protein,ERK1/2,JNK and p38MAPK in heart,lung and brain tissues decreased greatly after restoration of spontaneous circulation(P<0.05),cells and neurons in the CA1 region of heart,lung and hippocampus were severely damaged.Com-pared with cardiopulmonary resuscitation group,change trend of relevant indicators in electroacupuncture group was opposite to the above,damage of cells and neurons in rat heart,lung and brain tissue was reduced(P<0.05).αBGT reduced the anti-inflammatory effect of electroacupuncture on cardiopulmonary resuscitation rats(P<0.05).GTS-21 enhanced the anti-inflammatory protection of electroacupuncture on cardiopulmonary resuscitation rats(P<0.05).Conclusion:Electroacupuncture has anti-inflammatory and pro-tective effects on cardiopulmonary resuscitation rats based on CAP,and NF-κB/MAPKs participate in α7nAChR mediated CAP.

Purpose/Significance By using data mining methods,the medication rules and prescription characteristics of traditional Chinese medicine(TCM)for treating hypertension with preserved ejection fraction heart failure are discussed.Method/Process The da-tabase is established based on the TCMprescription information of hypertensive patients with preserved ejection fraction heart failure ad-mitted to the Cardiovascular Disease Department of the First Affiliated Hospital of Heilongjiang University of Chinese Medicine from Janu-ary 2019 to July 2022.SPSS Statistics and Moderler software are used to analyze the frequency,taste,meridian tropism,efficacy,associ-ation rules,clustering and factor analysis of the prescriptions to explore medication rules.Result/Conclusion TCMin the treatment of hy-pertension with preserved ejection fraction heart failure is mainly to replenish qi and spleen,nourish the heart and calm the mind,relieve phlegm,relieve cough and asthma,and pay attention to promoting blood circulation and removing blood stasis,promoting water infiltra-tion and dampness,which can reflect the overall concept of TCMand the characteristics of diagnosis and treatment based on syndrome dif-ferentiation and provide references for clinical medication.

Abstract OBJECTIVE To explore the effect of the polymorphisms of ATM(rs11212617), KCNJ11(rs5219), CYP2C9(rs1799853, rs1057910), TCF7L2(rs12255372, rs290487) and IRS1(rs1801278) on efficacy of metformin and gliclazide combined treatment for diabetes mellitus type 2. METHODS Eighty-one patients with newly diagnosed diabetes mellitus type 2 in Standardized Metabolic Disease Management Center of the Endocrinology Department of of Zhejiang Chinese Medical University Affiliated Jiaxing TCM Hospital were enrolled in this study, and they were treated with metformin hydrochloride tablets and gliclazide modified release tablets. MassARRAY was used to type the above single nucleotide polymorphism(SNP), and followed up for 3 months. Fasting plasma glucose(FPG), glycosylated hemoglobin type Alc(HbA1c) and fasting insulin(FINS) were compared before and after treatment. RESULTS The gene frequencies of all SNPs were compliant to Hardy-Weinberg equilibrium, the samples were well represented. The rs1799853, rs1057910, rs12255372 and rs1801278 had low mutation frequencies with poor clinical significance. The benefits of FPG compliance rate, increase of FINS, decrease of HbAlc after treatment were stronger in patients with homozygous rs5219 C allele than those with T-allele carriers (P<0.05). However, there were no significant differences in treatment effects among the genotypes of rs11212617 and rs290487. CONCLUSION The rs5219 polymorphism of KCNJ11 gene is associated with the efficacy of metformin hydrochloride tablets and gliclazide modified release tablets in treating diabetes mellitus type 2, with a more significant effect observed in individuals homozygous for the C allele. This finding can serve as a reference for personalized medication in clinical treatment of diabetes mellitus type 2.

ObjectiveTo explore the effect of Anmeidan on the sleep quality and serum levels of brain-derived neurotrophic factor （BDNF）， glial fibrillary acidic protein （GFAP）， and irisin in the patients with chronic insomnia. MethodA multicenter， randomized， double-blind， placebo-controlled clinical study was carried out， including 480 patients with chronic insomnia （deficiency syndrome） in Wuhan （Hubei）， Guangzhou （Guangdong）， and Lanzhou （Gansu）. They were randomized into an observation group and a control group at a ratio of 1∶1. The observation group was orally administered with Anmeidan granules at a dose of 11 g， 3 times per day， and the control group with Anmeidan simulant at a dose of 11 g， 3 times per day， Both groups of patients received sleep education after enrollment. After 4 weeks of medication， the Athens insomnia scale （AIS） scores， Spiegel scale scores， and serum levels of BDNF， GFAP， and irisin were compared between the two groups as well as between before and after treatment. ResultA total of 480 adult patients with chronic insomnia were enrolled in this study， with 64 patients falled off. Finally， the 415 patients were included in the analysis， including 213 patients in the observation group and 202 patients in the control group. There was no difference in age or sex between the two groups of patients. Compared with before treatment， the treatment in both groups decreased the AIS and Spiegel scores （P<0.01）. After treatment， the observation group had lower AIS and Spiegel scores than the control group （P<0.01）. The treatment in the observation group slightly lowered the level of BDNF， elevated the level of irisin （P<0.05）， and lowered the level of GFAP （P<0.05） in the serum. After treatment， the observation group showed higher level of irisin （P<0.05） and lower levels of BDNF and GFAP in the serum than the control group. ConclusionAnmeidan may improve the sleep quality of patients with chronic insomnia by elevating the irisin level and lowering the GFAP level in the serum.

Objective:To identify the pathogenic gene and inheritance pattern in a pedigree of congenital iris coloboma with congenital cataract.Methods:The method of pedigree investigation was adopted.A pedigree of congenital iris coloboma with congenital cataract was collected by Yunnan Disabled Rehabilitation Center and the 2nd Afliated Hospital of Kunming Medical University in February 2020.Ophthalmic examinations were carried out on the female proband, her parents, her children and her husband, and the clinical diagnosis was made.Genomic DNA was extracted from peripheral blood samples collected from the family members.The suspected pathogenic gene in the proband and her husband was screened by whole exome sequencing and was identified by bioinformatics analysis.The amino acid conservation was analyzed by UGENE software.The impact of the mutation on protein translation was predicted using MutationTaster software.The pathogenicity of the mutation was assessed according to the American College of Medical Genetics (ACMG) Standards and Guidelines.Pathogenic gene and mutations were verified by Sanger sequencing.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of the 2nd Afliated Hospital of Kunming Medical University (No.PJ-2020-61).Written informed consent was obtained from each subject or custodian.Results:The proband showed large iris defects in both eyes with only a small amount of observable iris tissue in the periphery, lens cortical opacity and posterior capsule opacification, accompanied by nystagmus.A novel heterozygous frameshift variation c. 415dupA (p.R139fs) was located in exon 8 of PAX6 gene, and the variation was conservative across multiple species.The variation was in the highly conserved region of PAX6 gene and caused the dysfunction of PAX6 protein.The variation was graded as PVS1+ PM2+ PP1, a pathogenic variation, based on ACMG guidelines.The pedigree was consistent with co-segregation, indicating that the novel variation was pathogenic.The proband and her children were diagnosed, but her parents were phenotypically normal, in accordance with autosomal dominant inheritance. Conclusions:The novel frameshift variation c.415dupA (p.R139fs) on the exon 8 of PAX6 gene is responsible for congenital iris coloboma with congenital cataract in the pedigree.This is the first report of this novel variation in PAX6 gene.

Objective:To investigate the predictive value of acute gastrointestinal injury (AGI) score for the severity of acute pancreatitis (AP), infectious pancreatic necrosis and patients′ death.Methods:Clinical data of 719 patients with AP were collected from the AP database of the National Clinical Research Center for Digestive System Diseases from January 2016 to June 2018. According to the severity of the disease, they were divided into MAP group (506 cases), MSAP group (112 cases) and SAP group (101 cases). AGI, APACHEⅡ, MCTSI and BISAP scores were calcululated in the three groups. Receiver operating characteristic curve (ROC) was drawn and the area under the curve (AUC) was calculated. The predictive value of the above four scoring systems for the hospitalization days, disease severity, infectious pancreatic necrosis and death was analyzed, respectively.Results:There were no cases of infectious pancreatic necrosis or death in the MAP group, but there were 9 cases of infectious pancreatic necrosis and 2 deaths in the MSAP group and 19 cases of infectious pancreatic necrosis and 8 deaths in the SAP group. There was a strong correlation between AGI score and AP patients′ hospitalization days ( r=0.619). AUC of AGI, APACHEⅡ, MCTSI and BISAP score in predicting the AP patients′ severity (MSAP+ SAP) was 0.967 (95% CI 0.951-0.982), 0.769(95% CI 0.720-0.899), 0.842(95% CI 0.809-0.875), 0.862 (95% CI0.832-0.893). AUC for forecasting infectious pancreatic necrosis was 0.803, 0.677, 0.692, 0.724, and the 95% CI was 0.724-0.882, 0.573-0.781, 0.582-0.636, 0.801-0.812. AUC for predicting death in patients with AP were 0.915, 0.597, 0.659, 0.812, and the 95% CI were 0.843-0.986, 0.444-0.751, 0.498-0.698 and 0.882-0.926. AGI score had the highest predictive value, followed by BISAP score, and the correlation between these two scores was the closest. The predictive value of AGI combined with BISAP score for infectious pancreatic necrosis and patient death (AUC were 0.837, 0.942, 95% CI were 0.770-0.903, 0.897-0.987) was better than that of AGI and BISAP score alone. Conclusions:AGI score combined with BISAP score is more effective in predicting the severity of AP, the occurrence of infectious pancreatic necrosis or patient death.