Objective To observe the therapeutic effects of Fourteen-Needle Awakening Acupuncture combined with repetitive transcranial magnetic stimulation(rTMS)on post-stroke patients with minimally conscious state(MCS).Methods Sixty MCS patients admitted to the Rehabilitation Department of Guangdong Second Traditional Chinese Medicine Hospital between December 2023 and December 2024 were randomly divided into an observation group and a control group,with 30 cases in each group.Both groups received conventional awakening therapy.The control group received additional sham rTMS,while the observation group received Fourteen-Needle Awakening Acupuncture combined with rTMS.The treatment duration was 14 days for both groups.After two weeks,awakening status was evaluated.Changes in Coma Recovery Scale-Revised(CRS-R)scores,Glasgow Coma Scale(GCS)scores,Full Outline of Unresponsiveness(FOUR)scores,as well as serum levels of vascular endothelial growth factor(VEGF)and neuron-specific enolase(NSE)were compared between groups before and after treatment.Results(1)After treatment,17 patients in the observation group regained consciousness,with a recovery rate of 56.67％(17/30);9 patients in the control group regained consciousness,with a recovery rate of 30.00％(9/30).The recovery rate in the observation group was significantly higher than that in the control group(P＜0.05).(2)After treatment,the CRS-R scores in both groups were significantly improved(P＜0.05),with the observation group showing a greater degree of improvement(P＜0.05).(3)After treatment,the FOUR scores and GCS scores of both groups were significantly improved(P＜0.05),with the observation group showing a greater degree of improvement(P＜0.05).(4)After treatment,the serum VEGF and NSE levels of both groups were significantly improved(P＜0.05),with the observation group showing a greater degree of improvement(P＜0.05).Conclusion Fourteen-Needle Awakening Acupuncture combined with rTMS effectively enhances cortical excitability and modulates electrophysiological activity of cerebral neurons in post-stroke MCS patients.This combined therapy demonstrates positive therapeutic effects in improving MCS and significantly accelerates consciousness recovery.

Objective To investigate the therapeutic effects of varying treatment durations of recombinant human brain natriuretic peptide(rhBNP)on heart failure following percutaneous coronary intervention(PCI)in patients with acute ST-segment elevation myocardial infarction(STEMI).Methods A total of 196 STEMI patients with heart failure(HF)following emergency percutaneous coronary intervention(PCI)were enrolled and randomly assigned to one of four groups:control group(n=53),short-course rhBNP group(n=47),medium-course rhBNP group(n=50),and long-course rhBNP group(n=46).The rates of cardiovascular mortality and HF-related rehospitalization were evaluated at 30 days and 6 months post-treatment.Serum levels of N-terminal pro-B-type natriuretic peptide(NT-proBNP),matrix metalloproteinase-9(MMP-9),tissue inhibitor of matrix metalloproteinase-1(TIMP-1),left ventricular end-diastolic diameter(LVEDD),and left ventricular ejection fraction(LVEF)were measured at 24 hours,3 days,1 week,30 days,and 6 months after HF treatment initiation.Results Compared with the control group,both the short-and medium-term rhBNP groups showed a significant reduction in cardiovascular mortality and HF-related rehospitalization rates in the long-term rhBNP group at 30 days and 6 months(P<0.05).Additionally,the medium-term rhBNP group exhibited lower HF-related rehospitalization rates than both the control and short-term rhBNP groups(P<0.05).Serum levels of NT-proBNP,MMP-9,and LVEDD significantly decreased in the short-term rhBNP group within 24 hours compared to the control group(P<0.05),while TIMP and LVEF levels increased(P<0.05).In comparison with the short-term rhBNP group,the medium-term rhBNP group demonstrated sustained reductions in NT-proBNP,MMP-9,and LVEDD levels at 72 hours,1 week,30 days,and 6 months(P<0.05),accompanied by increases in TIMP and LVEF(P<0.05).Furthermore,the long-term rhBNP group showed greater improvements than the medium-term group,with significantly lower NT-proBNP,MMP-9,and LVEDD levels and higher TIMP and LVEF values at 1 week,30 days,and 6 months(P<0.05).In terms of the adverse reactions,the incidence of hypotension in the control group,short course rhBNP group,medium course rhBNP group and long course rhBNP group increased successively(P<0.05).Conclusion The clinical efficacy of rhBNP in STEMI patients with HF following PCI gradually improved as the treatment duration increased,but the incidence of hypotension also rose accordingly.

Objective:To explore the molecular mechanism by which the methionine adenosyltransferase 2A (MAT2A)/β-catenin/zinc finger E-box binding homeobox 1 (ZEB1)/epithelial-mesenchymal transition (EMT) pathway regulates neural tube defect (NTD) through intracellular S-adenosylmethionine (SAM).Methods:A mouse NTD model was induced using the SAM metabolic disorder inhibitor ethionine. Eighty specific pathogen-free C57BL/6 mice were divided into three groups: a normal group (36 mice), an ethionine group (46 mice), and an ethionine+SAM group (44 mice). Phosphate-buffered saline (PBS), ethionine, and ethionine+SAM were respectively injected intraperitoneally on embryonic day 7.5 (E7.5), and the mice were sacrificed on E10.5. Embryonic tissues were collected, and the morphology of embryos in each group was observed under a stereomicroscope. The interaction between ethionine and MAT2A was analyzed using Autodock software. The expression levels of MAT2A, β-catenin, ZEB1, and EMT-related proteins in the brain tissues of embryos from the three groups were measured using immunofluorescence, immunohistochemistry, Western blotting, enzyme-linked immunosorbent assay (ELISA), and real-time quantitative polymerase chain reaction (RT-qPCR). Variance analysis was used for intergroup comparisons.Results:(1) Autodock analysis results showed that MAT2A binds to ethionine through covalent bonds, exhibiting a complementary effect, thereby accelerating the expression of MAT2A. (2) After successful construction of the NTD model, normal embryos were plump with well-developed brains. NTD embryos showed delayed development, obvious anencephaly, unclosed neural tubes, and asymmetry. (3) The levels of SAM and SAH in the embryonic tissues of the ethionine group were significantly lower than those in the normal group (1 737.56±95.64 vs. 872.33±205.11, and 89.17±9.50 vs. 51.25±9.48, respectively). The SAM and SAH levels in the ethionine+SAM group was 1 197.00±222.27 and 66.61±12.25, significantly higher than those in the ethionine group ( P<0.017). Compared with the normal group and the ethionine+SAM group, the expression of MAT2A mRNA in the embryonic brain tissue of the ethionine group was significantly upregulated (1.00±0.00, 1.59±0.52, and 2.42±0.53, respectively, F=49.64, P<0.001; pairwise comparisons between groups P<0.017). (4) Compared with the normal group, the expression of Ctnnb1 in the ethionine group was reduced, and the expression of Ctnnb1 in the ethionine+SAM group was higher than that in the ethionine group (1.00±0.00, 0.38±0.16, and 0.76±0.10, respectively, F=149.03, P<0.001; pairwise comparisons between groups P<0.017). (5) The expression of ZEB1 in the ethionine group was higher than that in the normal group and the ethionine+SAM group (2.91±0.55, 1.00±0.00, and 1.61±0.20, respectively, F=150.01, P<0.001; pairwise comparisons between groups P<0.017). (6) The expression levels of E-cadherin and Vimentin in the ethionine group were lower than those in the normal group. In contrast, the expression of N-cadherin was higher than that in the normal group. After SAM supplementation, the expression levels of E-cadherin and Vimentin were upregulated, and the expression level of N-cadherin was downregulated (0.54±0.12, 1.00±0.00, and 0.72±0.14, respectively, F=87.44; 0.53±0.17, 1.00±0.00, and 0.76±0.09, F=87.44; 3.11±0.53, 1.00±0.00, and 2.13±0.56, F=95.54; all P<0.001; pairwise comparisons within the same index group P<0.017]). Conclusions:Ethionine promotes the expression of MAT2A, leading to reduced SAM production. Ethionine regulates the level of ZEB1 by increasing MAT2A and inhibits the EMT process to interfere with methionine cycle metabolism, ultimately resulting in NTD.

Objective To study the horizontal transfer mechanism of optrA gene-carrying linezolid-resistant En-terococcus faecalis(LREf)among clinical Enterococcus faecalis mediated by plasmids.Methods Non-repeated LREf from a tertiary first-class hospital in Kunming City from August 2022 to August 2023 were collected and iden-tified by matrix-assisted laser desorption/ionisation-time of flight mass spectrometry(MALDI-TOF MS).Antimi-crobial susceptibility testing was performed by VITEK 2 Compact,disc diffusion method and microbroth dilution method,optrA gene was detected by polymerase chain reaction(PCR),molecular biology characteristics of LREf was analyzed by whole-genome sequencing(WGS),LREf as the donor strain and clinically isolated Enterococcus faecalis as the recipient strain for conjugation test.Results A total of 17 LREf strains were collected,optrA gene was detected from plasmids of 12 LREf strains.Multiple resistance genes and virulence genes were detected.12 LREf strains were mainly ST16 type(50.0％).Among the 24 transconjugates in the conjugation test,8 were suc-cessfully conjugated,with a conjugation rate of 33.3％.Further analysis revealed that IS1216E insertion sequences presented at upstream and downstream of the optrA gene on the plasmid before and after conjugation of strains,and formed IS1216E-fexA-optrA-erm(A)-IS1216E-like transposon units.Conclusion The optrA gene can be horizon-tally transferred among clinical Enterococcus faecalis by mediating of plasmid of genes carrying both erm(A)and fexA,and the insertion sequence IS1216E plays an important role in its horizontal transfer process.

Objective:To analyze effects of T11TS in the treatment of Cryptococcus neoformans fungal infection and the regula-tion of T-cell calcineurin(CaN)-activated T lymphoid nuclear factor(NFAT)pathway.Methods:SD rats infected with Cryptococcus neoformans were treated with immunoenhancer T11TS.Flow cytometry,Western blot and nuclear translocation were used to detect the changes of T cell function and related signal pathway.Results:The number of Cryptococcus neoformans in lung and spleen of CT3 group was significantly less than Cryptococcus neoformans infection group,CT1 group and CT2 group(P<0.05).The results of flow cy-tometry showed that the expression levels of LCK,FYN,LAT,PLCγ1,CaN,NFAT and IL-2 in Cryptococcus neoformans infection group were significantly lower than control group(P<0.05).The expression levels of LCK,FYN,LAT,PLCγ1,CaN,NFAT and IL-2 of CT1,CT2 and CT3 groups were significantly higher than Cryptococcus neoformans infection group(P<0.05).The results of Western blot were similar to those of flow cytometry.Conclusion:T11TS in the treatment of Cryptococcus neoformans infection can enhance the immunity of the body by up-regulating the expression of related signal factors in the CaN-NFAT pathway of T lymphocytes,increasing the expression of NFAT and IL-2,stimulating the activation and proliferation of T lymphocytes.

Objective:To analyze effects of T11TS in the treatment of Cryptococcus neoformans fungal infection and the regula-tion of T-cell calcineurin(CaN)-activated T lymphoid nuclear factor(NFAT)pathway.Methods:SD rats infected with Cryptococcus neoformans were treated with immunoenhancer T11TS.Flow cytometry,Western blot and nuclear translocation were used to detect the changes of T cell function and related signal pathway.Results:The number of Cryptococcus neoformans in lung and spleen of CT3 group was significantly less than Cryptococcus neoformans infection group,CT1 group and CT2 group(P<0.05).The results of flow cy-tometry showed that the expression levels of LCK,FYN,LAT,PLCγ1,CaN,NFAT and IL-2 in Cryptococcus neoformans infection group were significantly lower than control group(P<0.05).The expression levels of LCK,FYN,LAT,PLCγ1,CaN,NFAT and IL-2 of CT1,CT2 and CT3 groups were significantly higher than Cryptococcus neoformans infection group(P<0.05).The results of Western blot were similar to those of flow cytometry.Conclusion:T11TS in the treatment of Cryptococcus neoformans infection can enhance the immunity of the body by up-regulating the expression of related signal factors in the CaN-NFAT pathway of T lymphocytes,increasing the expression of NFAT and IL-2,stimulating the activation and proliferation of T lymphocytes.

Objective To investigate the therapeutic effects of varying treatment durations of recombinant human brain natriuretic peptide(rhBNP)on heart failure following percutaneous coronary intervention(PCI)in patients with acute ST-segment elevation myocardial infarction(STEMI).Methods A total of 196 STEMI patients with heart failure(HF)following emergency percutaneous coronary intervention(PCI)were enrolled and randomly assigned to one of four groups:control group(n=53),short-course rhBNP group(n=47),medium-course rhBNP group(n=50),and long-course rhBNP group(n=46).The rates of cardiovascular mortality and HF-related rehospitalization were evaluated at 30 days and 6 months post-treatment.Serum levels of N-terminal pro-B-type natriuretic peptide(NT-proBNP),matrix metalloproteinase-9(MMP-9),tissue inhibitor of matrix metalloproteinase-1(TIMP-1),left ventricular end-diastolic diameter(LVEDD),and left ventricular ejection fraction(LVEF)were measured at 24 hours,3 days,1 week,30 days,and 6 months after HF treatment initiation.Results Compared with the control group,both the short-and medium-term rhBNP groups showed a significant reduction in cardiovascular mortality and HF-related rehospitalization rates in the long-term rhBNP group at 30 days and 6 months(P<0.05).Additionally,the medium-term rhBNP group exhibited lower HF-related rehospitalization rates than both the control and short-term rhBNP groups(P<0.05).Serum levels of NT-proBNP,MMP-9,and LVEDD significantly decreased in the short-term rhBNP group within 24 hours compared to the control group(P<0.05),while TIMP and LVEF levels increased(P<0.05).In comparison with the short-term rhBNP group,the medium-term rhBNP group demonstrated sustained reductions in NT-proBNP,MMP-9,and LVEDD levels at 72 hours,1 week,30 days,and 6 months(P<0.05),accompanied by increases in TIMP and LVEF(P<0.05).Furthermore,the long-term rhBNP group showed greater improvements than the medium-term group,with significantly lower NT-proBNP,MMP-9,and LVEDD levels and higher TIMP and LVEF values at 1 week,30 days,and 6 months(P<0.05).In terms of the adverse reactions,the incidence of hypotension in the control group,short course rhBNP group,medium course rhBNP group and long course rhBNP group increased successively(P<0.05).Conclusion The clinical efficacy of rhBNP in STEMI patients with HF following PCI gradually improved as the treatment duration increased,but the incidence of hypotension also rose accordingly.

Objective To study the horizontal transfer mechanism of optrA gene-carrying linezolid-resistant En-terococcus faecalis(LREf)among clinical Enterococcus faecalis mediated by plasmids.Methods Non-repeated LREf from a tertiary first-class hospital in Kunming City from August 2022 to August 2023 were collected and iden-tified by matrix-assisted laser desorption/ionisation-time of flight mass spectrometry(MALDI-TOF MS).Antimi-crobial susceptibility testing was performed by VITEK 2 Compact,disc diffusion method and microbroth dilution method,optrA gene was detected by polymerase chain reaction(PCR),molecular biology characteristics of LREf was analyzed by whole-genome sequencing(WGS),LREf as the donor strain and clinically isolated Enterococcus faecalis as the recipient strain for conjugation test.Results A total of 17 LREf strains were collected,optrA gene was detected from plasmids of 12 LREf strains.Multiple resistance genes and virulence genes were detected.12 LREf strains were mainly ST16 type(50.0％).Among the 24 transconjugates in the conjugation test,8 were suc-cessfully conjugated,with a conjugation rate of 33.3％.Further analysis revealed that IS1216E insertion sequences presented at upstream and downstream of the optrA gene on the plasmid before and after conjugation of strains,and formed IS1216E-fexA-optrA-erm(A)-IS1216E-like transposon units.Conclusion The optrA gene can be horizon-tally transferred among clinical Enterococcus faecalis by mediating of plasmid of genes carrying both erm(A)and fexA,and the insertion sequence IS1216E plays an important role in its horizontal transfer process.

Objective:To explore the molecular mechanism by which the methionine adenosyltransferase 2A (MAT2A)/β-catenin/zinc finger E-box binding homeobox 1 (ZEB1)/epithelial-mesenchymal transition (EMT) pathway regulates neural tube defect (NTD) through intracellular S-adenosylmethionine (SAM).Methods:A mouse NTD model was induced using the SAM metabolic disorder inhibitor ethionine. Eighty specific pathogen-free C57BL/6 mice were divided into three groups: a normal group (36 mice), an ethionine group (46 mice), and an ethionine+SAM group (44 mice). Phosphate-buffered saline (PBS), ethionine, and ethionine+SAM were respectively injected intraperitoneally on embryonic day 7.5 (E7.5), and the mice were sacrificed on E10.5. Embryonic tissues were collected, and the morphology of embryos in each group was observed under a stereomicroscope. The interaction between ethionine and MAT2A was analyzed using Autodock software. The expression levels of MAT2A, β-catenin, ZEB1, and EMT-related proteins in the brain tissues of embryos from the three groups were measured using immunofluorescence, immunohistochemistry, Western blotting, enzyme-linked immunosorbent assay (ELISA), and real-time quantitative polymerase chain reaction (RT-qPCR). Variance analysis was used for intergroup comparisons.Results:(1) Autodock analysis results showed that MAT2A binds to ethionine through covalent bonds, exhibiting a complementary effect, thereby accelerating the expression of MAT2A. (2) After successful construction of the NTD model, normal embryos were plump with well-developed brains. NTD embryos showed delayed development, obvious anencephaly, unclosed neural tubes, and asymmetry. (3) The levels of SAM and SAH in the embryonic tissues of the ethionine group were significantly lower than those in the normal group (1 737.56±95.64 vs. 872.33±205.11, and 89.17±9.50 vs. 51.25±9.48, respectively). The SAM and SAH levels in the ethionine+SAM group was 1 197.00±222.27 and 66.61±12.25, significantly higher than those in the ethionine group ( P<0.017). Compared with the normal group and the ethionine+SAM group, the expression of MAT2A mRNA in the embryonic brain tissue of the ethionine group was significantly upregulated (1.00±0.00, 1.59±0.52, and 2.42±0.53, respectively, F=49.64, P<0.001; pairwise comparisons between groups P<0.017). (4) Compared with the normal group, the expression of Ctnnb1 in the ethionine group was reduced, and the expression of Ctnnb1 in the ethionine+SAM group was higher than that in the ethionine group (1.00±0.00, 0.38±0.16, and 0.76±0.10, respectively, F=149.03, P<0.001; pairwise comparisons between groups P<0.017). (5) The expression of ZEB1 in the ethionine group was higher than that in the normal group and the ethionine+SAM group (2.91±0.55, 1.00±0.00, and 1.61±0.20, respectively, F=150.01, P<0.001; pairwise comparisons between groups P<0.017). (6) The expression levels of E-cadherin and Vimentin in the ethionine group were lower than those in the normal group. In contrast, the expression of N-cadherin was higher than that in the normal group. After SAM supplementation, the expression levels of E-cadherin and Vimentin were upregulated, and the expression level of N-cadherin was downregulated (0.54±0.12, 1.00±0.00, and 0.72±0.14, respectively, F=87.44; 0.53±0.17, 1.00±0.00, and 0.76±0.09, F=87.44; 3.11±0.53, 1.00±0.00, and 2.13±0.56, F=95.54; all P<0.001; pairwise comparisons within the same index group P<0.017]). Conclusions:Ethionine promotes the expression of MAT2A, leading to reduced SAM production. Ethionine regulates the level of ZEB1 by increasing MAT2A and inhibits the EMT process to interfere with methionine cycle metabolism, ultimately resulting in NTD.

ObjectiveTo observe the distribution of traditional Chinese medicine （TCM） syndrome types in advanced pancreatic cancer patients， and explore the association between median survival time and different TCM syndromes and different intervention times of Chinese herbal medicine （CHM）. MethodsThe clinical data of 136 advanced pancreatic cancer patients who have received CHM for more than 3 months were collected retrospectively， including gender， age， family history， smoking history， drinking history， location of disease， lymph node metastasis， multiple distant metastasis， western medicine treatment methods， TCM diagnosis and treatment information， and survival time. The Kaplan-Meier （KM） estimator was used， and the median survival time of patients was calculated. The TCM syndrome type of each patient was judged， and the main single syndrome types and compound syndrome types were summarized. The median survival time was compared among different compound syndrome types. The patients were further divided into the group of those having received CHM ≥6 months and those having received CHM ＜6 months. Whether receiving CHM ≥6 months was taken as the grouping variable， while the matching variables were age， gender， family history， smoking history， drinking history， location of disease， lymph node metastasis， multiple distant metastasis， surgery， chemotherapy， and radiotherapy when propensity score matching was performed， and the difference in median survival time between the two groups of patients before and after matching was compared. ResultsFor 136 cases of advanced pancreatic cancer， the top five single syndromes were spleen qi deficiency， liver blood stasis， liver qi stagnation， spleen dampness， and liver heat. The main compound types were liver constraint， spleen deficiency and blood stasis syndrome， liver-gallbladder damp-heat and blood stasis syndrome， liver constraint， qi stagnation and spleen deficiency syndrome， spleen-stomach yang deficiency and blood stasis syndrome， and spleen deficiency and dampness-heat internal accumulation syndrome. The overall median survival time before and after matching was 12.47 （7.70，17.10） months and 13.77 （8.83，17.20） months， respectively， and was significantly higher in the group treated with CHM ≥ 6 months than that treated with CHM ＜6 months （P＜0.05）. Among the 136 patients before matching， the median survival time of patients with spleen deficiency and dampness-heat internal accumulation syndrome was longest ［16.23 （14.17，19.40） months］， while that of patients with spleen-stomach yang deficiency and blood stasis syndrome was the shortest ［7.33 （5.80，12.83） months］. For patients with liver constraint， spleen deficiency and blood stasis syndrome， liver-gallbladder damp-heat and blood stasis syndrome， and spleen-stomach yang deficiency and blood stasis syndrome， those having received CHM ≥ 6 months have much longer median survival time than those having received CHM ＜6 months （P＜0.05）. Among the 108 patients after matching， the median survival time of those with spleen deficiency and dampness-heat internal accumulation syndrome was the longest ［15.23 （7.67，18.27） months］， while that of spleen-stomach yang deficiency and blood stasis syndrome was the shortest ［8.80 （6.90，16.17） months］. For patients with liver-gallbladder dampness-heat and blood stasis syndrome and spleen-stomach yang deficiency and blood stasis syndrome， the median survival time was higher in the group treated with CHM ≥ 6 months treated with CHM ＜6 months （P＜0.05）. ConclusionAfter treatment with CHM， advanced pancreatic cancer patients with spleen deficiency and damp-heat internal accumulation had a better prognosis， while those with spleen-stomach yang deficiency and blood stasis had a worse prognosis. Treatment with CHM ≥ 6 months could extend the median survival of advanced pancreatic cancer patients with liver-gallbladder damp-heat and blood stasis syndrome and spleen-stomach yang deficiency and blood stasis syndrome.