Objective To analyze the MTHFR C677T gene polymorphism and homocysteine in women in early pregnancy in Ordos region,to clarify the distribution characteristics of MTHFR C677T gene polymor-phism and the correlation between the two,and to provide genetic basis for scientific guidance on folic acid supplementation during pregnancy and prevention of birth defects.Methods A total of 602 Han women in early pregnancy who were registered and underwent early pregnancy examinations in the gynecology clinic of Ordos Central Hospital from September 2022 to September 2023 were selected as the research subjects.Blood samples were collected from all research objects.MTHFR C677T gene polymorphism was detected by using PCR chip hybridization,and homocysteine level was detected by biochemical enzyme circulation method.Sta-tistical analysis of MTHFR C677T locus genotype and allele frequency,as well as their correlation with homo-cysteine was conducted.Results The detection frequencies of MTHFR C677T gene polymorphism CC,CT,and TT types were 23.6%,47.5%,and 28.9%,respectively.The detection frequencies of alleles C and T were 47.3%and 52.7%,respectively.There were statistically significant differences compared to Han women in Shanghai,Wenzhou,Meishan,Nanning,and other regions(P<0.05),but there was no statistically significant difference compared to Han women in Xi'an(P>0.05).The serum homocysteine level of pregnant women with TT genotype was higher than that of pregnant women with CC and CT genotypes,while the serum ho-mocysteine level of pregnant women with CT genotype was higher than that of pregnant women with CC gen-otype(P<0.05).The CT and TT genotypes of MTHFR C677T were both risk factors for hyperhomocys-teinemia in women in early pregnancy in this region,the risk was 2.80 and 8.07 times higher than that of the CC genotype,respectively,and the differences were statistically significant(P<0.05).Conclusion The distri-bution of MTHFR C677T gene polymorphism Han women in early pregnancy in Ordos region has regional characteristics and is correlated with homocysteine level.Developing personalized folic acid supplementation plans based on different genotypes during pregnancy is of great significance for preventing birth defects.

ObjectiveTo investigate the effect of rutin on the browning of 3T3-L1 preadipocytes and the mechanism. MethodCell counting kit-8 （CCK-8） assay was used to detect the effect of different concentration of rutin （3.125， 6.25， 12.5， 25， 50， 100， 200 μmol·L^-1） on 3T3-L1 cell activity， and Western blot to examine the effect of rutin （12.5， 25， 50 μmol·L^-1） on the expression of thermogenesis-associated proteins uncoupling protein 1 （UCP1）， PR domain containing 16 （PRDM16） and peroxisome proliferator-activated receptor γ coactivator-1α （PGC-1α） in adipocytes. After the optimal concentration of rutin was determined， the effect of rutin on lipid droplet formation in adipocytes was observed based on oil red O staining， and the expression of nuclear respiratory factor 1 （NRF1）， nuclear respiratory factor 2 （NRF2） and mitochondrial transcription factor A （TFAM）， which were the landmark proteins of mitochondrial biosynthesis， was detected by Western blot. ResultCompared with the blank group， 200 μmol·L^-1 rutin inhibited 3T3-L1 cell activity （P<0.01）. Compared with the blank group， at the concentration of 12.5， 25， 50 μmol·L^-1 rutin significantly promoted the expression of thermogenesis-associated proteins （UCP1， PRDM16， and PGC-1α） （P<0.01）， which was determined as the optimal concentration. Compared with the blank group， 50 μmol·L^-1 rutin significantly increased the immunofluorescence intensity of mitochondrial UCP1 protein in 3T3-L1 cells （P<0.01） and the expression of the markers of mitochondrial biosynthesis （NRF1， NRF2， and TFAM） （P<0.01）. In addition， 50 μmol·L^-1 rutin significantly inhibited lipid droplet formation of 3T3-L1 adipocytes （P<0.01）. ConclusionRutin inhibited lipid droplet deposition in 3T3-L1 adipocytes and increased the expression of thermogenesis-related proteins （UCP1， PRDM16， and PGC-1α） and markers of mitochondrial biosynthesis （NRF1， NRF2， and TFAM）， thereby inducing the browning of 3T3-L1 adipocytes. This lays a basis for the development of drugs that safely regulate the browning of white cells.

Mulberry (Morus alba L.) leaf is a well-established traditional Chinese botanical and culinary resource. It has found widespread application in the management of diabetes. The bioactive constituents of mulberry leaf, specifically mulberry leaf flavonoids (MLFs), exhibit pronounced potential in the amelioration of type 2 diabetes (T2D). This potential is attributed to their ability to safeguard pancreatic β cells, enhance insulin resistance, and inhibit α-glucosidase activity. Our antecedent research findings underscore the substantial therapeutic efficacy of MLFs in treating T2D. However, the precise mechanistic underpinnings of MLF's anti-T2D effects remain the subject of inquiry. Activation of brown/beige adipocytes is a novel and promising strategy for T2D treatment. In the present study, our primary objective was to elucidate the impact of MLFs on adipose tissue browning in db/db mice and 3T3-L1 cells and elucidate its underlying mechanism. The results manifested that MLFs reduced body weight and food intake, alleviated hepatic steatosis, improved insulin sensitivity, and increased lipolysis and thermogenesis in db/db mice. Moreover, MLFs activated brown adipose tissue (BAT) and induced the browning of inguinal white adipose tissue (IWAT) and 3T3-L1 adipocytes by increasing the expressions of brown adipocyte marker genes and proteins such as uncoupling protein 1 (UCP1) and beige adipocyte marker genes such as transmembrane protein 26 (Tmem26), thereby promoting mitochondrial biogenesis. Mechanistically, MLFs facilitated the activation of BAT and the induction of WAT browning to ameliorate T2D primarily through the activation of AMP-activated protein kinase (AMPK)/sirtuin 1 (SIRT1)/peroxisome proliferator-activated receptor-gamma coactivator 1α (PGC-1α) signaling pathway. These findings highlight the unique capacity of MLF to counteract T2D by enhancing BAT activation and inducing browning of IWAT, thereby ameliorating glucose and lipid metabolism disorders. As such, MLFs emerge as a prospective and innovative browning agent for the treatment of T2D.
Mice ; Animals ; Adipose Tissue, Brown ; Sirtuin 1/pharmacology* ; Diabetes Mellitus, Type 2/metabolism* ; AMP-Activated Protein Kinases/metabolism* ; Morus/metabolism* ; Flavonoids/metabolism* ; Prospective Studies ; Signal Transduction ; Adipose Tissue, White ; Plant Leaves ; Uncoupling Protein 1/metabolism* ; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha/metabolism*

The incidence of diabetes has been on the rise as the result of lifestyle changes， especially the high-fat diet and reduced exercise. Thus， it has become a global public health problem and it is an urgent task to explore effective therapy. There has been an explosion of research on the relationship of transforming growth factor-β （TGF-β） signaling pathways with diabetes complications and tumors， but the role of the pathways in the occurrence and progression of diabetes remains unclear. TGF-β signaling pathways can be activated by many factors， directly or indirectly leading to the apoptosis of islet β cells and insulin resistance （IR）， and thus they are expected to become new targets for the treatment of diabetes. TGF-β-related signaling pathways involve AMP-activated proteinkinase （AMPK）， protooncogene （c-Myc）， Ski-relatednovel protein N （SnoN）， Smad ubiquitination regulatory factor 1 （Smurf1）， miR-335-5p， and other signaling molecules. They participate in the occurrence and development of IR， apoptosis of islet β cells， insulin secretion disorder， fibrosis of adipocytes， and metabolic disorder of adipocytes， and inhibit the browning of white adipose tissue， playing an important part in the pathological process of human diabetes. According to traditional Chinese medicine （TCM）， the pathogenesis of diabetes is the deficiency of Qi and Yin， and the late stage is characterized by the syndrome of Qi deficiency， and Yang deficiency and blood stasis， which should be treated according to the principle of replenishing Qi and nourishing Yin， warming Yang and activating blood. It has been found that the efficacy of some Chinese medicinals and compound prescriptions on diabetes is closely related to the TGF-β signaling pathways. This paper reviews TGF-β-associated signaling pathways， elucidating the roles of them in pathogenesis of diabetes， and analyzes the relationship of TGF-β-associated signaling pathways with the effect of compound Chinese medicine prescriptions against diabetes. This study is expected to lay a theoretical basis for the research on the treatment diabetes.

ObjectiveTo investigate the medicinal effect of total flavonoids of mulberry leaves on regulating liver lipid metabolism disorder in diabetes mellitus type 2 （T2DM） rats， and the mechanism based on liver peroxidase proliferators activate receptors-α （PPAR-α） and carnitine palmityl transferase-1 （CPT-1） proteins. MethodTotal flavonoids of mulberry leaves were extracted and purified by ethanol extraction + macroporous resin purification and then identified. T2DM rat model was induced by high fat diet （HFD） + streptozocin（STZ）method. Rats with blood glucose ≥ 11.1 mmol·L^-1 were divided into three administration groups with the high dose （300 mg·kg^-1）， medium dose （150 mg·kg^-1）， and low dose （75 mg·kg^-1） of total flavonoids of mulberry leaves for 8 weeks， respectively， to observe the weight and blood glucose of the rats. The pathological changes of rat livers were observed by hematoxylin-eosin （HE） staining. Biochemical method was used to detect the levels of total cholesterol （TC）， triglyceride （TG）， low density lipoprotein-cholesterol （LDL-C）， and high density lipoprotein-cholesterol （HDL-C） of blood lipid metabolism in rats. The messenger ribonucleic acid （mRNA） and protein expressions of PPAR-α and CPT-1 were determined by real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot. ResultAfter 8 weeks of intervention of total flavonoids of mulberry leaves， compared with the control group， the food intake， liver index， and fasting blood glucose of rats in the model group increased significantly （P<0.01）. Compared with the model group， the food intake， fasting blood glucose， and liver index of rats in the administration groups decreased significantly （P<0.01）. The results of HE staining showed that the liver tissue structure of rats in the control group was complete and there was no obvious abnormality. The model group showed vacuolar degeneration and inflammatory infiltration of hepatocytes of rats. There was no obvious abnormality in the liver structure of rats in the administration groups. The results of blood lipid showed that compared with the control group， the levels of TC， TG， and LDL-C increased significantly （P<0.01）， but the level of HDL-C decreased significantly （P<0.01） in the model group. Compared with the model group， the levels of TC， TG， and LDL-C decreased significantly （P<0.05， P<0.01）， whereas the level of HDL-C increased significantly （P<0.01） in the administration groups. The results of Real-time PCR showed that compared with the control group， the mRNA expression of PPAR-α and CPT-1 of rats in the model group decreased significantly （P<0.01）. Compared with the model group， the mRNA expressions of PPAR-α and CPT-1 of rats in the high-dose group increased significantly （P<0.01）. The results of Western blot showed that compared with the control group， the protein expressions of PPAR-α and CPT-1 of rats in the model group decreased significantly （P<0.01）. Compared with the model group， the protein expressions of PPAR-α and CPT-1 of rats in the high-dose group increased significantly （P<0.05， P<0.01）. ConclusionTotal flavonoids of mulberry leaves can effectively reduce blood glucose and improve liver lipid metabolism disorder in T2DM rats. The total flavonoids of mulberry leaves could regulate lipid metabolism and play a hypoglycemic role by activating and regulating PPAR-α and CPT-1 proteins and promoting oxidative decomposition of fatty acids.

Objective:To observe the relationship between the different stages of type 2 diabetes mellitus(T2DM)and the intestinal flora and verify its underlying mechanism.Methods:T2DM rats were generated by high-fat diet(HFD)combined with intraperitoneal streptozo-tocin(STZ)injection.The rats were divided into four groups:the control group(fed with normal feed for 1 month),the HFD group(fed with HFD for 1 month),the T2DM group(HFD combined with STZ and blood glucose＞11.1 mM),and the unformed T2DM model(Un-mod)group(HFD combined with STZ and blood glucose＜11.1 mM).Feces were collected,and bacterial communities in the fecal samples were analyzed by 16S rRNA gene sequencing.The content of short-chain fatty acids(SCFAs)in feces was measured by gas chromatography.Western blot and quantitative real-time polymerase chain reaction were used to detect the expression of G protein-coupled receptor 41(GPR41)and GPR43.Results:At different stages of T2DM,the intestinal flora and SCFAs content of rats were significantly decreased(all P＜.05).Our results indicated that g_Prevotella had a significant negative correlation,and g_Ruminococcus_torques_group and g_lachnoclastic had a significant positive correlation with blood glucose.The content of SCFAs,in particular acetate and butyrate,in rat feces of different stages of T2DM were significantly reduced,as well as GPR41 and GPR43 expression.The results in the Un-mod group were similar to the T2DM group,and the expression of GPR41 and GPR43 proteins were significantly higher than those in the T2DM group(both P＜.001).Conclusion:The intestinal flora-SCFAs-GPR41/GPR43 network may be important in the development of T2DM.Decreasing blood glucose levels by regulating the intestinal flora may become a new therapeutic strategy for T2DM,which has very important clinical and social values.

Objective:To investigate the changes of non-specific and HBV core antigen (HBcAg)-specific Th9 cells, and intereleukin-9 (IL-9) in HBV-infected patients, and to assess the influence of Th9 cells on CD8 + T cell function. Methods:Twelve patients with acute hepatitis B (AHB) and 58 with chronic hepatitis B (CHB), who were hospitalized in the First Affiliated Hospital of Xinxiang Medical University between January 2018 and January 2019, were enrolled in this study. Twenty healthy subjects negative for HBsAg were selected as controls. Peripheral blood mononuclear cells (PBMCs) and plasma samples were isolated. Non-specific Th9 cells (CD3 + CD4 + IL-9 + ) and HBcAg-specific Th9 cells were analyzed by flow cytometry. Plasma IL-9 level was measured by enzyme linked immunosorbent assay. CHB patients received tenofovir disoproxil fumarate (TDF) antiviral therapy. The changes of non-specific Th9 cells, HBcAg-specific Th9 cells and plasma IL-9 level were assessed 48 weeks after TDF therapy. CD4 + CCR4 -CCR6 -CXCR3 -(Th9) cells and CD8 + T cells were isolated from 12 HLA-A2 restricted CHB patients and co-cultured with HepG2.2.15 cells with the presence of anti-IL-9 neutralizing antibody. The percentage of dead HepG2.2.15 cells and the levels of IFN-γ and TNF-α were detected. Student′s t test, one-way analysis of variance or SNK- q test was used for statistical comparison between groups. Results:There were no significant differences in non-specific Th9 cells or plasma IL-9 level among AHB patients, CHB patients and healthy controls ( P>0.05). HBcAg-specific Th9 cells was down-regulated in CHB patients when compared with AHB patients [(2.49±0.61)% vs (3.19±0.62)%, P<0.001]. The percentage of HBcAg-specific Th9 cells was negatively correlated with HBV DNA ( r=-0.385, P=0.003), but not correlated with ALT ( P>0.05) in CHB patients. TDF therapy for 48 weeks remarkably elevated the HBcAg-specific Th9 cells [(2.94±0.48)%, P<0.001], however, did not affect non-specific Th9 cells or plasma IL-9 level ( P>0.05) in CHB patients. The cytotoxicity of HBcAg-specific Th9 cells was low in CHB patients. However, HBcAg-specific Th9 cells could induce enhanced cytotoxicity of CD8 + T cells to HepG2.2.15 cells, which manifested as increased percentage of dead HepG2.2.15 cells and higher levels of IFN-γ and TNF-α. Anti-IL-9 neutralizing antibody reduced the enhancement of CD8 + T cell cytotoxicity by HBcAg-specific Th9 cells ( P<0.001). Conclusions:Chronic HBV infection might suppress the level and function of HBcAg-specific Th9 cells, resulting in persistent infection.

The widespread application of next generation sequencing (NGS) in clinical settings has enabled testing, diagnosis, treatment and prevention of genetic diseases. However, many issues have arisen in the meanwhile. One of the most pressing issues is the lack of standards for reporting genetic test results across different service providers. The First Forum on Standards and Specifications for Clinical Genetic Testing was held to address the issue in Shenzhen, China, on October 28, 2017. Participants, including geneticists, clinicians, and representatives of genetic testing service providers, discussed problems of clinical genetic testing services across in China and shared opinions on principles, challenges, and standards for reporting clinical genetic test results. Here we summarize expert opinions presented at the seminar and report the consensus, which will serve as a basis for the development of standards and guidelines for reporting of clinical genetic testing results, in order to promote the standardization and regulation of genetic testing services in China.

Objective To investigate the types of staphylococcal cassette chromosome mec (SCCmec)gene and an-timicrobial resistance of community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA)isolated from outpatients and inpatients in a hospital.Methods MRSA strains isolated between May 2011 and August 2015 in a hospi-tal and the relevant case data were collected,polymerase chain reaction(PCR)method was used to identify mecA gene of MRSA and SCCmec gene of CA-MRSA,antimicrobial susceptibility testing of CA-MRSA were performed and analyzed. Results A total of 305 MRSA isolates were collected,296 of which were mecA positive,29.73% (88/296)were CA-MR-SA. The genotyping of CA-MRSA showed that 48 strains were SCCmec type Ⅳ,36 were SCCmec type V,the other 4 strains were undefined. Antimicrobial susceptibility testing results showed that susceptibility rates of CA-MRSA to vanco-mycin,linezolid,and tigecycline were all 100% ,resistance rates to penicillin and oxacillin were both 100% ;resistance rates of SCCmec type IV and SCCmec type V CA-MRSA strains to levofloxacin,rifampicin,and ciprofloxacin were all signifi-cantly different (all P<0.05),to ampicillin/sulbactam,furantoin,and erythromycin were all >58% .Conclusion The main SCCmec type of CA-MRSA are type IV and type V in this hospital,antimicrobial resistance rate is high,clinicians should pay high attention,and use antimicrobial agents according to antimicrobial susceptibility testing results.

Objective To analyze antimicrobial resistance of hospital-associated methicillin-resistant Staphylococ-cusaureus(HA-MRSA)and community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA),and provide reference for clinical treatment and rational antimicrobial use. Methods From May 2013 to June 2014, Staphylococcus aureus in a hospital were collected and analyzed,strains were identified and performed antimicrobial susceptibility testing by using VITEK 2 Compact system,diagnosis of HA-MRSA and CA-MRSA were confirmed in combined with clinical symptoms.Results A total of 84 MRSA isolates were isolated (61 were HA-MRSA strains,23 were CA-MRSA).Resistant rates of HA-MRSA and CA-MRSA to penicillin G and oxacillin were both 100.00% ;to ampicillin/sulbactam was 100.00% and 95.65% respectively;to compound sulfamethoxazole was 39.34% and 34.78% respectively. Antimicrobial resistant rates of HA-MRSA to gentamicin,tetracycline,erythro-mycin,clindamycin,levofloxacin,ciprofloxacin,moxifloxacin,nitrofurantoin,and rifampicin were all higher than CA-MRSA,the difference were significant(all P<0.001).Conclusion Antimicrobial resistance of HA-MRSA and CA-MRSA are all serious,monitor should be intensified,antimicrobial use should be chosen according to antimicro-bial susceptibility testing result.