Objective: To investigate changes in lactate and histone lactylation during senescence of mouse bone marrow mesenchymal stem cells (BMSCs), to clarify the effects of regulating lactate metabolism on the osteogenic differentiation of senescent BMSCs, and to provide theoretical evidence and an experimental basis for metabolic regulatory interventions in aging-related osteoporosis and impaired bone repair. Methods: Approval was obtained from the Institutional Laboratory Animal Ethics Committee. BMSCs were isolated using the whole bone marrow adherence method and surface markers (CD29, CD90, CD11b and CD45) and identified by flow cytometry. A cellular model of BMSCs senescence was established using different concentrations of etoposide (0, 5, 10, 15, and 20 μmol/L). The gene and protein expression levels of senescence-related markers, namely, p53, p21, and p16, were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot analysis, and the optimal induction concentration was determined using CCK-8 and SA-β-gal assays. BMSCs were divided into a control group and an etoposide-induced senescence group to examine lactate production and the expression of histone lactylation-related enzymes, including Ldha, Ep300, and Sirt2. Western blot analysis was used to analyze pan-lysine lactylation（Pan-Kla） and histone H3 lysine 18 lactylation （H3K18la). During the lactate production inhibition experiments, BMSCs were divided into three groups: control, senescence group (treated with 10 μmol/L etoposide), and senescence+ GNE-140 group (treated with 10 μmol/L etoposide and 1 μmol/L GNE-140). qRT-PCR, Western blot, and SA-β-gal staining were performed to assess the effects of lactate production inhibition on cellular senescence phenotypes. During osteogenic differentiation, BMSCs were assigned to a control group, senescence group (treated with 10 μmol/L etoposide), senescence+ 5 mmol/L sodium lactate group, and senescence+ 10 mmol/L sodium lactate group. The levels of Pan-Kla and H3K18la were detected by Western blot. The mRNA expressions of runt-related transcription factor 2 (Runx2), transcription factor Sp7 (Sp7) and alkaline phosphatase (Alpl) were detected by qRT-PCR. The protein expressions of collagen type I (COL1), RUNX2, osterix (OSX) and osteocalcin (OCN) were detected by Western blot, and mineralized nodule formation was observed by Alizarin Red S staining. Results: Flow cytometry analysis indicated that the isolated BMSCs highly expressed CD29 (97.3%) and CD90 (95.7%). Etoposide induced BMSC senescence by activating the p53-p21 signaling pathway, with an optimal concentration of 10 μmol/L. Compared with the control group, BMSCs exhibited decreased lactate production, reduced mRNA expression of Ldha and Ep300 (P<0.001), upregulated Sirt2 expression (P<0.01), and diminished levels of Pan-Kla and H3K18la modification (P<0.001). Further inhibition of lactate production with GNE 140 further increased the expression levels of p53 and p21 and the percentage of SA β gal positive cells (P<0.01). In osteogenic differentiation experiments, the expression levels of Runx2, Sp7, Alpl mRNA and COL1, RUNX2, OSX, OCN proteins, as well as the mineralized nodule formation ability of senescent BMSCs, were decreased (P<0.05); exogenous supplementation with 5 mmol/L and 10 mmol/L sodium lactate can increase the levels of panolactic lactation and histone lactation, and improved the osteogenic differentiation capacity and mineralized nodule formation of senescent BMSCs. Conclusion BMSC senescence is associated with reduced lactate production and decreased histone lactylation levels. Exogenous supplementation with sodium lactate partially rescues osteogenic differentiation impairment in senescent BMSCs.

OBJECTIVE To evaluate the quality of evidence in the systematic evaluation/meta-analysis of traditional Chinese medicine （TCM） for diabetes retinopathy （DR） based on the GRADE system. METHODS Chinese and English databases were searched to obtain the relevant studies of systematic evaluation/meta-analysis of traditional Chinese medicine in the treatment of DR. The search time was from the establishment of each database to January 13th， 2024. According to the inclusion and exclusion criteria， literature screening was conducted. After extracting relevant information from the included literature， the GRADE system was used to evaluate the quality level of the evidence body in the included studies， and the evidence of the outcome indicators was integrated and summarized. RESULTS A total of 51 studies were ultimately included， encompassing 135 outcome indexes. Among these， 19 indicators （14.1%） were of high quality， 87 （64.4%） were of medium quality， 26 （19.3%） were of low quality， and 3 （2.2%） were of very low quality. Overall， the evidence quality of the outcome indicators in the included studies was medium to low quality. The integrated results of evidence on the efficacy of outcome indexes showed that compared with conventional Western medicine， calcium dobesilate or placebo， TCM had significant advantages in improving overall efficacy， reducing bleeding spot area， reducing macular foveal thickness， and increasing visual improvement rate. In addition，the combination of TCM and conventional Western medicine or calcium dobesilate was significantly more effective than using conventional Western medicine or calcium dobesilate alone. CONCLUSIONS The overall quality of the evidence in the systematic evaluation/meta-analysis study on the treatment of DR with TCM is medium to low quality. Based on existing research findings， TCM demonstrates good clinical efficacy in the treatment of DR.

AIM:To investigate the effects of the γ-aminobutyric acid(GABAergic)neural pathway from the anterior cingulate cortex(ACC)to the nucleus accumbens(NAc)on the regulation of irritable bowel syndrome(IBS)and its underlying mechanisms in mice.METHODS:(1)A C57BL/6J mice model of IBS was established by using chronic acute combing stress(CACS).The mice were divided into a normal group and an IBS group(n=8).The presence of IBS-like symptoms was determined through behavioural tests,an intestinal motility test and abdominal withdrawal reflex scores.(2)Fluorescence gold(FG)retrograde tracing and immunohistochemistry were used to investigate the ACC-NAc GABAergic neural pathway and to examine the activation of GABA in the ACC in IBS mice(n=8).(3)A total of 1.5 μL of normal saline(NS),GABAA receptor antagonist bicuculline(BIC)or agonist isoguvacine hydrochloride(Isog)was ad-ministered via a preburied catheter into the NAc of mice in IBS and normal groups.The mice were randomly divided into three groups(n=8):NS group,BIC group and Isog group.IBS-like symptoms were assessed.(4)The mice were prein-jected with AAV2/9-mDlx-iCre-WPRE-pA in the ACC and AAV2/2Retro Plus-hSyn-DIO-hM3D(Gq)-eGFP-WPRE-pA in the NAc and subsequently divided into four groups(n=8):NS(intraperitoneal injection)+NS(NAc microinjection)group,NS+BIC group,clozapine N-oxide(CNO)+NS group and CNO+BIC group.The mice who received AAV2/2Retro-hSyn-DIO-hM4D(Gi)-EGFP-WPRE-pA in the NAc were randomly divided into three groups(n=8):NS+NS group,NS+BIC group and CNO+NS group.Enzyme-linked immunosorbent assay(ELISA)was employed to estimate the expression levels of histamine and 5-hydroxytryptamine(5-HT)in colon tissue,and the effects of GABAergic neural pathways from ACC to NAc on IBS were studied.RESULTS:CACS induced IBS-like symptoms in mice.The results of FG retrograde tracing combined with fluorescence immunohistochemistry showed that GABA neurons of ACC could project to NAc.The injection of BIC in the NAc was found to significantly reduce anxiety-like behaviours,diarrheal symptoms and visceral hy-persensitivity in the IBS mice(P<0.05).Chemogenetic inhibition of the ACC-NAc GABAergic neurons ameliorated IBS-like symptoms in mice(P<0.05).CONCLUSION:The GABAergic pathway of ACC-NAc might be involved in the regu-lation of IBS in mice,which may be related to the release of histamine and 5-HT in colon tissue.

Objective To analyze the clinical efficacy of preoperative short-course radiotherapy combined with neoadjuvant chemotherapy in locally advanced mid-low rectal cancer.Methods Eighty patients with locally advanced mid-low advanced rectal cancer were enrolled and assigned to single group(preoperative neoadjuvant chemotherapy,n=32)and combination group(preoperative short-course radiotherapy+neoadjuvant chemotherapy,n=48)according to different treatment methods.All of them underwent total mesorectal resection after chemoradiotherapy.The two groups were compared for clinical curative effect,serum tumor markers[carcinoembryonic antigen(CEA),carbohydrate antigen 125(CA125),CA199]and immune function(CD3+,CD4+,CD8+)before and after chemoradiotherapy;and the incidence of adverse reactions during chemoradiotherapy was recorded.Results The overall remission rate was higher in combination group than in single group(37.50%vs 15.63%,P<0.05).After chemoradiotherapy,the levels of serum CEA,CA125 and CA199 in both groups were decreased,and they were lower in combination group than in single group(P<0.05).After chemoradiotherapy,the levels of CD3+and CD4+in single group and combination group increased,and the level of CD8+decreased(P<0.05),but there was no statistically significant differences in the above indicators between two groups(P>0.05).Additionally,the difference in adverse reactions between two group was trivial(P>0.05),and no grade-IV adverse reactions occurred in either group.Conclusion The combination of preoperative short-course radiotherapy and neoadjuvant chemotherapy is effective for locally advanced mid-low advanced rectal cancer,and beneficial for increasing overall remission rate and lowering the levels of serum tumor markers,with high safety.

Objective To analyze the clinical efficacy of preoperative short-course radiotherapy combined with neoadjuvant chemotherapy in locally advanced mid-low rectal cancer.Methods Eighty patients with locally advanced mid-low advanced rectal cancer were enrolled and assigned to single group(preoperative neoadjuvant chemotherapy,n=32)and combination group(preoperative short-course radiotherapy+neoadjuvant chemotherapy,n=48)according to different treatment methods.All of them underwent total mesorectal resection after chemoradiotherapy.The two groups were compared for clinical curative effect,serum tumor markers[carcinoembryonic antigen(CEA),carbohydrate antigen 125(CA125),CA199]and immune function(CD3+,CD4+,CD8+)before and after chemoradiotherapy;and the incidence of adverse reactions during chemoradiotherapy was recorded.Results The overall remission rate was higher in combination group than in single group(37.50%vs 15.63%,P<0.05).After chemoradiotherapy,the levels of serum CEA,CA125 and CA199 in both groups were decreased,and they were lower in combination group than in single group(P<0.05).After chemoradiotherapy,the levels of CD3+and CD4+in single group and combination group increased,and the level of CD8+decreased(P<0.05),but there was no statistically significant differences in the above indicators between two groups(P>0.05).Additionally,the difference in adverse reactions between two group was trivial(P>0.05),and no grade-IV adverse reactions occurred in either group.Conclusion The combination of preoperative short-course radiotherapy and neoadjuvant chemotherapy is effective for locally advanced mid-low advanced rectal cancer,and beneficial for increasing overall remission rate and lowering the levels of serum tumor markers,with high safety.

Objective To explore the occurrence of fetal chromosomal abnormalities among pregnant women with an adverse reproductive history using traditional karyotyping and single nucleotide polymorphism microarray(SNP-array)technology.Methods Totally 94 in 2 163(4.35%)cases of singleton pregnant women with an adverse reproductive history were performed amniocentesis in Jinhua Maternal and Child Health Care Hospital from June 2015 to June 2017. Traditional karyotyping and SNP-array were employed simultaneously for prenatal diagnosis,and the detection rates of the two methods were compared. Results All of the 94 specimens were successfully analyzed, 11 cases were found with chromosomal anomaly, the overall detection rate was 11.7%(11/94). Seven (7.4%,7/94) abnormalities cases were detected by karyotyping,and 7(7.4%)by SNP-array.The karyotyping results of trisomy 21,and 45,X and the deletion of chromosome 13 were consistent with SNP-array.Only 3(3.2%,3/94)microdeletion/duplications (the sizes of duplications and deletions were between 422.4-1 708.4 kb)and 1(1/4)loss of heterozygosity were detected by SNP-array,but were missed by karyotyping.Furthermore, 2 cases′copy number variation were found pathogenic gene related, while the other 2 were considered benign or variant of uncertain significance. Four cases(4/7)of abnormalities were detected by karyotyping, while confirmed balanced translocation and inversion by SNP-array.All patients were informed and chosen to continue the pregnancy.Conclusions The rate of abnormal fetal chromosomes in pregnant women with an adverse reproductive history is still high.SNP-array is a new molecular genetic technique,and combined with use of traditional karyotyping,it could improve the detection rate of fetal chromosomal abnormalities and reduce abortion rate, thus providing a basis for genetic counseling and prenatal diagnosis.

ObjectiveTo evaluate totally laparoscopic Meckel's diverticulectomy in comparison with laparoscopic-assisted or open diverticulectomy.MethodsThe clinical data of 58 cases of Meckel's diverticulum admitted between January 2006 and January 2011 were analyzed.Cases were divided into three groups according to different period of time. As a result,totally laparoscopic surgery was performed in 13 cases,laparoscopic-assisted resection in 25 cases and open diverticulectomy in 20 cases. Totally laparoscopic group was compared with the other two groups in operation time,flatus defecation time,the incidence of postoperative complications and postoperative hospital stay.ResultsThe mean length of incision was ( 1.6 ± 0.4 ) cm,the mean operation time was ( 41 ± 5 ) min,flatus defecation time was (21.2 ±3.7) h,and the postoperative hospital stay was (6.3 ± 1.2) d in totally laparoscopic group.While that was (2.5 ± 1.2 ) m,( 38 ± 2 ) cm,( 23.6 ± 4.2 ) h,( 6.5 ± 2.3 ) d,respectively in laparoscopicassisted group,and the mean length of incision was (5.0 ± 2.2 ) cm,the mean operation time was (51 ± 6 )min,flatus defecation time was (32.3 ± 6.7) h,the postoperative hospital stay was (8.4 ± 3.8) d in open surgery group.Compared with conventional laparotomy,laparoscopic techniques enjoy advantages of minimal invasion,shorter operative time,fewer complications,shorter recovery period and earlier gastrointestinal recovery(P ＜ 0.05). There were nosignificant differences in operative time, recovery period and complications between totally laparoscopic group and laparoscopic-assisted group.ConclusionsTotally laparoscopic Meckel's diverticulectomy is safe,effective and miniinvasive in experienced hands.