Cardiac arrhythmia is one of the most common cardiovascular diseases,with high morbidity and mortality,which threatens human health and lives. With the development of medicine, natural products are revealing ever-greater anti-arrhythmic benefits and potential. However,their targets have not been fully clarified. In the recent ten years ,scientists have been studying the molecular mechanisms of natural products that have been found to inhibit INa,ICa-L,Ito,IK1,IKr,IKM3,HERG channel current and steady state K+ current,promote IKs and IKATP,inhibit microRNA-1 expression and change cardiac microRNA expression profile,affect Na+-K+-ATPase and superoxide dismutase activity,inhibitβ receptor and angiotensinⅡ receptor,and regulate lipid metabolism,thus affecting cardiac rhythm and exerting anti-arrhythmic effect. This paper revies the research advances in the antiarrhythmic tar?gets of natural products.

To improve the inulinase application in biotechnology, the characteristic of inulinase from Kluyveromyces marxianus YX01 was investigated. The inu gene of K. marxianus YX01 was transformed into Pichiapastoris GS115 host cells with molecular biology techniques. Then we achieved the heterologous expression of exo-inulinase whose molecular mass was about 86.0 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Furthermore, six His-tag was added to the inulinase and a two-step method was applied in the purification of inulinase, including concentration via dialysis by polyethylene glycol 20 000 and metal Ni-NTA Agarose affinity adsorption. The purification factor of purified protein was 3.6 and the recovery rate of enzyme activity was 33.1%. We characterized the purified inulinase. The optimum temperature was 60 degrees C and pH was 4.62. When inulin and sucrose were used as substrates, the K(m) and V(max) values were 80.53 g/L vs 4.49 g/(L x min) and 183.10 g/L vs 20.20 g/(L x min), respectively. In addition, metal ions including Mn2+, Ca2+, Cu2+, Zn2+ and Fe2+ exhibited different degrees of inhibition on the enzyme activity, and Cu2+, Zn2+ and Fe2+ exhibited the most significant inhibition. Our findings might lay a good foundation for industrial application of inulinase.
Glycoside Hydrolases ; chemistry ; genetics ; Industrial Microbiology ; Inulin ; Kluyveromyces ; enzymology ; genetics ; Pichia ; Sucrose ; Temperature

Objective To investigate the relationship between the methylation of CpG island of RUNX3 gene promoter and its expression in a human cutaneous malignant melanoma cell line A375, and to assess the role of RUNX3 gene methylation in the pathogenesis of human cutaneous malignant melanoma. Methods Cultured A375 cells were treated with various concentrations (0, 1, 5, 10, 20 μmol/l) of 5-azacyti-dine for 24 or 72 hours followed by another 5 days of culture. Then, methylation-specific PCR (MSP) was performed to evaluate the methylation status of RUNX3 promoter region, and Western-blot analysis to detect the protein expression of RUNX3 in A375 cells. Results The RUNX3 gene promoter region was hypermethylated in untreated A375 cells, along with the absence of protein expression of RUNX3. However, after the treatment with 5-azacytidine, the promoter region of RUNX3 gene was demethylated partly, and the expression of RUNX3 protein was restored in A375 cells. Further, the expression intensity was directly correlated with the concentration of 5-azacytidine. Conclusions The promoter hypermethylation of RUNX3 gene may be related to the silencing of RUNX3 gene expression in A375 cells, whereas 5-azacytidine can cause the demethylation of RUNX3 gene, reactivate the gene which has been inactivated by the promoter hypermethylation, and finally induce the re-expression of RUNX3 protein.

Objective In order to investigate the synaptic plasticity in dentate gyrus after seizures and axonal and dendrtic sprout- ing induced by KA administration. Methods The density of synapses, the curvature forms of synaptic interface were studied under electron microscope. Results 1 .The density of the synapse is decreased obviously 3 days after KA injection, while the density of synapses is increased to control level 7 days after KA injection. 2. Compare with the curvation forms of synaptic interface of control animals and 3days after KA administration animals, the amount of smile synapses is significantly decresed and the amount of frown synapses is significantly increased in the moleculous layer of dentate gyrus 7 days after KA injection. Conclusions 1 .This result demonstrates that the axonal and dendritic sprouting of dentate granule cells is functional. 2. The increase of frown synapse is related to the release of glutamate of sprouting mossy fibers.

Objective To study the effects and molecular mechanism of MAWBP in the proliferation, invasion and migration of human colorectal cancer cell lines. Methods Colorectal cancer cell lines, caco-2 and HT-29, were divided into MAWBP over-expressed group and control group. MAWBP was stablely over-expressed in both cell lines by lentivirus, and the positive clones were screened by puromycin. The expression of MAWBP gene at mRNA and protein levels was evaluated by RT-PCR and western blot. Cell proliferation was detected by MTT. Cell migration and invasion were detected by transwell experiment. The EMT related protein levels of Snail, E-cadherin, and MAPK pathway protein of ERK, p38, p-ERK, p-p38 were determined by western blot analysis. Results Compared with the control group, the expression of MAWBP was significantly higher in HT-29-MAWBP and caco2-MAWBP group . The cell proliferation , invasion and migration ability significantly reduced in the HT-29-MAWBP group. The expression of Snail, p-ERK and p-p38 significantly decreased, while the E-cadherin increased in HT-29-MAWBP and caco2-MAWBP group. Conclusions Over- expression of MAWBP exerts an inhibitory effect on the proliferation, invasion and migration of colorectal caner cells via inhibit the MAPK pathway and regulate the EMT process.

Objective To investigate the effect of phorbol-12-myristate-13-acetate(PMA)on cyclooxygenase-2(COX-2) mRNA and protein expression in cultured human HaCaT keratinocytes,and the mechanism for cytotoxity of PMA against keratinocytes.MethodsRT-PCR and Westem blot were utilized to detect the expression of COX-2 mRNA and protein in cultured HaCaT ells at 24 hours after the treatment with various concentrations of PMA (0.1,1.0,10 mg/L).ResultsWithout any treatment,there was no or a weak expression df COX-2 mRNA and protein in HaCaT cells;incubation witll PMA resulted in the induction of the expression of COX-2 in HaCaT cells.The expression levels of COX-2 mRNA and protein in 10 mg/L PMA-pretreated HaCaT cells were significantly higher than those in 1.0 mg/L PMA-pretreated HaCaT cells,which was in turn higher than that in 0.1 mg/L PMA-pretreated cells and untreated cells;the difrerence was statistically significant (all P＜0.01).Conclusion These results suggest that PMA may be involved in keratinocyte tumorigenesis by upregulating he expression of COX-2 as well as synthesis and release of prostaglandin in keratinocytes.

Objective To evaluate the clinical efficacy and safety of combined ozone hydrotherapy for the treatment of atopic dermatitis(AD). Methods A total of 60 patients with moderate or severe AD aged from 6 to 65 years were enrolled, and randomly and equally divided into a test group and a control group. Both the two groups were treated with oral levocetirizine capsules 5 mg once a day, topical tacrolimus ointment twice a day, and topical moisturizers. The test group was additionally treated with ozone hydrotherapy 3- 5 times every week. The treatment lasted 2 weeks. The severity scoring of atopic dermatitis (SCORAD) score, visual analog scale (VAS) score, dermatology life quality index (DLQI) or children′s dermatology life quality index (CDLQI) score were assessed before and after the treatment, and compared between the two groups. Enzyme?linked immunosorbent assay(ELISA) was performed to measure the levels of interleukin?4(IL?4)and nerve growth factor(NGF)in peripheral blood from the patients before and after the treatment. Results After 2?week treatment, the SCORAD scores, VAS scores and DLQI/CDLQI scores significantly decreased from 42.13 ± 16.03, 7.14 ± 2.12 and 14.92 ± 5.94 before the treatment to 27.3 ± 11.01, 2.23 ± 1.31 and 9.69 ± 4.17 respectively in the test group(all P<0.05), and from 43.36 ± 17.78, 6.45 ± 1.29 and 15.15 ± 5.40 to 39.65 ± 16.67, 3.32 ± 0.87 and 12.84 ± 5.37 respectively in the control group(all P<0.05). In addition, all the three scores were significantly lower in the test group than in the control group after 2?week treatment(all P<0.05). A significant decline was also observed in the levels of IL?4 and NGF in peripheral blood in the two groups at the end of the treatment compared with those at baseline(all P<0.05), and the magnitude of decline in IL?4 levels was significantly higher in the test group than in the control group(t=8.57, P<0.05), while that in NGF levels was insignificantly different between the two groups(t=9.51, P>0.05). Conclusion Combined ozone hydrotherapy can effectively and safely improve the condition of patients with AD, likely by decreasing the levels of IL?4 in peripheral blood.

Objective To investigate the viability and safety of laparoscopic sleeve gastrectomy(LSG)based on the TJ point at the junction of the posterior gastric wall and the apex of the medial edge of the left diaphragm.Methods A retrospective analysis of 135 patients with obesity or obesity with metabolic syndrome who underwent LSG from January 2019 to January 2022 were divided into two groups according to the different free modes of fundogastric body.68 patients underwent surgery using the TPOL model LSG.A control group of 67 patients was treated with conventional LSG surgery.To analyze and compare the duration of operation,gastric fundus free time,intraoperative blood loss time,hospital stay time,postoperative gastric fistula,bleeding rate,and occurrence of gastroesophageal reflux disease(GERD)between the two groups.Results All patients successfully completed LSG surgery and were safely discharged.They were followed for a period of 12 to 36 months.the duration of operation for the study group was(56.13±10.56)minutes,while for the control group it was(62.45±12.74)minutes.The gastric fundus was freed in(6.34±4.16)minutes for the study group and(12.58±6.37)minutes for the control group.The duration of hospitalization was(3.84±0.42)days for the study group and(4.06±0.69)days for the control group.The intraoperative blood loss was(10.87±1.28)ml for the study group and(15.56±3.39)ml for the control group.The incidence of postoperative GERD was 5(7.35％)for the study group and 13(19.40％)for the control group.The differences between the two groups were statistically significant(P＜0.05).The decrease in excess weight at 12 months after surgery,as well as complications such as bleeding and gastric fistula,were not statistically significant(P＞0.05).Conclusion LSG(TPOL model)extends from the TJ point(the Tri-junction point where the gastric left mesangium,gastric posterior mesangium,and pancreatic mesangium meet)to the apex of the medial foot edge of the left diaphragm.LSG(TPOL model)can be efficiently and safely achieve tension-free anastomosis,resulting in a complete free gastric fundus.This method is feasible,and has clinical value for the standardized free gastric fundus of LSG.

Traditional Chinese medicines have demonstrated clinical efficacy in preventing and treating chemotherapy-induced peripheral neuropathic pain(CIPNP).However,their specific clinical application and mechanism of action require further in-depth study and exploration.There is thus a need to develop more accurate and clinically relevant animal models that reflect the occurrence and development of human diseases as a tool for research.This review provides an in-depth analysis and discussion of the recent establishment and detection criteria of existing rat and mouse animal models of paclitaxel-induced peripheral neuropathic pain.We also evaluate and explain the application of these models for the prevention and treatment of CIPNP in traditional Chinese medicine,thus providing a theoretical basis and reference for future experimental and mechanistic research on the subject.This research will benefit clinical practice and promotion,offering valuable insights into preventing and treating CIPNP using traditional Chinese medicines.

Most of the CHD patients reveal string pulse, mainly due to damage of heart function, lowering of arterial compliance and increase of total peripheral resistance. The common pulse in patients of blood diseases reveal frequent, tiny, string and slippery characteristic, mostly due to the increase of compensatory pumping action of the heart, shortening of ejection time of the left heart, with better vessel compliance and hemorheology, low total peripheral resistance.