Objective To observe the clinical efficacy of Dulongneedling in the treatment of vertigo in cervical spondylosis of vertebral artery(CSA).Methods A total of 46 cases of patients with definitive diagnosis of vertigo in CSA admitted to the outpatient clinic of the Acupuncture,Muxibustion and Rehabilitation Department of Guangdong Second Traditional Chinese Medicine Hospital from June to October 2024 were selected for the study.The patients were randomly divided into observation group and control group according to the random number table method,with 23 cases in each group.The observation group was treated with Dulongten-needling in shoulder and neck,and the control group was given conventional acupuncture treatment,the course of treatment for the two groups covered two weeks.After two weeks of treatment,the clinical efficacy of the two groups was evaluated,and the changes in the scores of Evaluation Scale For Cervical Vertigo(ESCV)before and after treatment were observed in the patients of the two groups.Three dimensional pseudo-continuous arterial spin labeling(3D-pCASL)technique was applied to determin the changes in cerebral blood flow(CBF)values in the region of region of interest(ROI),and the changes in CBF values in the patients of the two groups were compared.Results(1)The total effective rate of the observation group was 91.30%(21/23),and that of the control group was 78.26%(18/23),and the efficacy of the observation group was superior to that of the control group,the difference being statistically significant(P<0.05).(2)After treatment,the ESCV scores of patients in the two groups were significantly improved(P<0.05),and the improvement in the observation group was significantly superior to that in the control group,the difference being statistically significant(P<0.05).(3)After treatment,the CBF change values of each ROI in the two groups showed elevated(positive),and the CBF change values of the right middle cerebral artery(R-MCA)in the control group showed decreased(negative).After treatment,the CBF change values of the observation group in the blood supply areas of the right posterior inferior cerebellar artery(R-PICA),right anterior inferior cerebellar artery(R-AICA),right anterior cerebral artery(R-ACA),branches from vertebral arteries(VA)and basilar arteries(BA),bilateral superior cerebellar arteries(SCA),and bilateral posterior cerebral arteries(PCA)were significantly elevated,and the observation group was significantly superior in improving the above ROIs in the CBF change value to that of the control group,the difference being statistically significant(P<0.05).Conclusion Dulongneedling in the treatment of vertigo in CSA can significantly improve the CBF of patients,reduce the clinical symptoms such as vertigo,neck and shoulder pain,headache.

Objective: To determine the expression of melanoma-associated antigens D4(MAGED4) and SIRT7 in human glioma, and to analyze the potential effects of MAGED4 and SIRT7 on glioma cell proliferation. Methods: The MAGED4 and SIRT7 expression levels and their correlation were compared by the China glioma genome atlas(CGGA), human protein atlas(HPA), and UALCAN databases. Survival analysis, ROC curve analysis, and Cox regression analysis were used to predict the outcome of MAGED4 and SIRT 7 in glioma patients. Gene ontology(GO) and Kyoto encyclopedia of genes and genomes(KEGG) signaling pathway enrichment analysis were used to explore the biological functions of MAGED4 and SIRT7 in glioma. Western blot experiment was used to investigate whether MAGED4 protein exerted its regulatory effects on the activity of the PI3K/AKT signaling pathway via SIRT7. The effect of MAGED4 on cell proliferation in glioma through SIRT7 was explored by CCK-8. Results: The analysis results of CGGA, UALCAN, and HPA databases showed that the expression levels of MAGED4 and SIRT7 in glioma tissues were higher than those in normal brain tissue, and the expression were positively correlated. Results of survival, ROC, and Cox analysis showed that high expression of MAGED4 and SIRT7 mRNA were risk factors for poor prognosis in glioma. Results of KEGG enrichment analysis showed that MAGED4 and SIRT7 were associated with the PI3K/AKT signaling in glioma, and Western blot results showed that MAGED4 activated the PI3K/AKT signaling pathway by regulating SIRT7. The CCK-8 results showed that MAGED4 promotes the proliferation of glioma cells through SIRT7. Conclusion MAGED4 and SIRT7 are highly expressed in glioma and associated with poor prognosis, and MAGED4 promotes glioma cell proliferation through activation of the PI3K/AKT signaling pathway by SIRT7.

Objective To investigate the effects of DNA demethylation drugs combined with histone deacetylase in-hibitors on fragile X mental retardation 1 neighbor protein (FMR1NB) expression and its promoter methylation in human oral cancer cells and try to find a strategy of weakening the heterogeneity of FMR1NB expression.Methods Human oral cancer cell lines Cal27 and SCC-9 were treated with decitabine (DAC) , an inhibitor of DNA meth-yltransferase, combined with trichostatin A (TSA) and valproic acid (VPA), inhibitors of histone deacetylase.Then reverse transcription-polymerase chain reaction (RT-PCR) , quantitative real-time PCR (qRT-PCR) and Western blot were used to detect the expression of FMR1 NB and pyrosequencing was used to detect the methylation of FMR1NB promoter.Results Compared with the blank control group, DAC and its combination with TSA and VPA significantly induced the expression of FMR1NB mRNA and protein in Cal27 and SCC-9 cells.Compared with DAC alone group, FMR1NB mRNA expression of each DAC-combined drug groups significantly increased, but FMR1NB protein did not significantly change in Cal27 cells; for SCC-9 cells, except for DAC+TSA group, the mRNA and protein levels of FMR1NB significantly increased in all other groups.In addition, there was no signifi-cant difference in the expression of FMR1 NB mRNA and protein between the three-combined drugs group and two-combined drugs groups.Further methylation assay showed that the methylation level of the overall FMR1NB promot-er and its each CpG site measured were reduced to varying degrees in all treatment groups except for three-combina-tion drug group of SCC-9.Conclusion DAC and its combination with TSA and VPA can enhance the expression of FMR1NB by mediating the demethylation of FMR1NB promoter, wherein the enhanced expression effect of the com-bination of the two drugs is stronger, suggesting that they have the potential to weaken the heterogeneity of FMR1NB expression and improve the immunotherapy effect of oral cancer.

Purpose To verify the safety and effectiveness of the Insight NM/CT Pro SPECT/CT Imaging System in clinical applications is verified through clinical validation of the acceptable rate of clinical image quality,commonly use features of the device,ease of use of the machine,satisfaction with the stability of the entire machine and safety evaluation.Materials and Methods A total of 79 subjects who voluntarily participated in the trial from July to October 2022 in the Affiliated Hospital of Inner Mongolia Medical University,were selected to undergo endocrine,circulatory,skeletal,respiratory and urological imaging for multiple systems using the Insight NM/CT Pro SPECT/CT,respectively.The quality of the images,the machine operation and safety were evaluated.Results The image quality of all 79 patients were evaluated.Of 36 cases,there were 35 cases of planar imaging were acceptable;Of 43 cases,there were 42 cases of tomographic imaging were acceptable,with an acceptability rate of 97.22%and 97.67%,respectively,and a lower limit of 95%CI of 85.47%and 87.71%,respectively,which were both higher than those of the target value(83.00%).When the operators performed imaging operations on 79 patients,the common functions of the equipment,the ease of use of the equipment and the stability of the whole machine were evaluated as a satisfaction rate of 100%,with no adverse events.Conclusion Insight NM/CT Pro SPECT/CT effectiveness fully meets the requirements of clinical applications,with clear images of tissue structure,full range of commonly used equipment features,stable system,and superior safety and reliability.

Objective To visually analyze the Chinese and English literature in the research field of Cistanches Herba through bibliometrics;To understand the research status and hotspots of Cistanches Herba;To provide reference for the related research.Methods Literature related to Cistanches Herba in CNKI and Web of Science core databases from January 1,2002 to December 31,2022 was retrieved.VOSviewer 1.6.18 was used for co-occurrence clustering and temporal overlay analysis of authors and keywords,while CiteSpace 6.1.R2 was used for keyword burst analysis.Results Totally 1 631 articles were included,including 1 481 Chinese articles and 150 English articles,and the overall number of publications was on the rise.This field has formed research teams represented by Guo Yuhai from Zhejiang Sci-Tech University,Tu Pengfei from Beijing University of Chinese Medicine,and Chen Jun and Xu Rong from Chinese Academy of Medical Sciences.High frequency keywords included Cistanches Herba,echinoside,phenylethanolside,high performance liquid chromatography,cistanche polysaccharide,artificial cultivation,anti-fatigue,etc.Conclusion The research in this field mainly focuses on TCM therapy,clinical study,etiology and pathogenesis.Network pharmacology,molecular biology,data mining and so on may become the focus of future research of Cistanches Herba.

Objective To detect the expression profile of heme oxygenase-1(HO-1)during the process of wound repair in radiation-wound combined injury(R-W-CI),and evaluate its wound healing improving effects of R-W-CI by HO-1 knockdown with siRNA.Methods A total of 36 male C57BL/6J mice(8 weeks old)were randomly and equally divided into a simple skin wound group(W group)and a skin wound group combined with whole-body radiation(6 Gy)injury(R-W-CI group).During the wound healing process,the wounds were photographed and recorded,and the residual areas were quantified by Image J.Wound tissues were sampled and stained with HE staining for pathological and histological observation,and the damage to the hematopoietic system was assessed by dynamic examination of the peripheral blood.The expression and changes of HO-1 in wound tissues were detected by q-PCR and Western blotting.Then,26 male C57BL/6J mice(8 weeks old)were randomly and equally divided into siRNA knockdown HO-1 group(si-HO-1 group)and siRNA negative control group(si-NC group).After radiation combined injury was inflicted,60 μL of F127 gel loaded with si-HO-1(5 μm/L)was applied to each wound in the si-HO-1 group,and an equal amount of F127 gel loaded with negative control si-NC was applied to the wound in the si-NC group.The knockdown of HO-1 in wound tissues was detected by Western blotting,and the changes in wound area were observed.In the wound tissues harvested in 3 d after wounding,the expression of cytokines IL-1β,IL-6 and TNF-α was examined by q-PCR and the proliferation of granulation tissues was evaluated by Ki67 immunohistochemical staining.HE staining was performed on wound tissues on day 3 and day 9 post-injury to assess the improvement effect of knockdown of HO-1 on wound healing of radiation combined injuries.Results Compared with the W group,semi-quantitative analysis of the residual wound area showed that healing was significantly delayed in the R-W-CI group on days 7 and 10 post-injury(P＜0.01).HE staining on day 7 showed that in the R-W-CI group,the re-epithelialization was delayed,and the growth of granulation tissues was poor;and at the same time,peripheral blood leukocytes and their classified counts showed a significant decrease in the early period after injury(P＜0.05).Further tests indicated that the expression of HO-1 protein was slightly higher in the wound of the R-W-CI group than that of the W group in 3 and 7 d after injury,though no significant difference(P＞0.05),whereas statistical difference was seen in 10 d(P＜0.05),accompanied by the distribution of the full-length and truncated forms of HO-1 protein.Quantitative PCR obtained similar results in the mRNA expression of HO-1 in wounds in both 7 and 10 d after injury(P＜0.05).siRNA intervention could effectively knock down the HO-1 protein level of the wounds(P＜0.05),promote wound contraction(P＜0.05),reduce the width of the wound(P＜0.01),up-regulate the inflammatory cytokines IL-6 and TNF-α in 3 d,enhance the proliferation of repair cells in wound margin,and improve the growth of the granulation tissue in the R-W-CI model when compared with the conditions after si-NC intervention.Conclusion There exists a sustained high expression level of HO-1 during wound repair,and wound knockdown of HO-1 by siRNA can improve the lack of inflammation status and promote wound healing in R-W-CI mice.

Objective: To investigate the contamination of phthalic acid esters (PAEs) and assess the health risk of PAEs contamination in market-available yellow rice wine in Huzhou City, Zhejiang Province, so as to provide the safety safeguard for consuming yellow rice wine. Methods: Yellow rice wine samples were collected from markets in Huzhou City from 2021 to 2022, and 16 PAEs were determined in yellow rice wine using magnetic solid-phase extraction coupled with gas chromatography-tandem mass spectrometry. The carcinogenic and non-carcinogenic risks of PAEs were evaluated using the health risk models proposed by United States Environmental Protection Agency. Results: A total of 75 yellow rice wine samples were collected, and 44 samples were detected with PAEs contamination (58.67%). Dimethyl phthalate (DMP), di-isobutyl phthalate (DIBP) and di-butyl phthalate (DBP) were detected, and there were 17 samples (22.67%) detected with DBP overdose (DMP and DIBP had no limit standard). DMP, DBP and DIBP, which were not classified as Class 2B and higher carcinogens by WHO's International Agency for Research on Cancer, had no definitive carcinogenic risks. Under mean PAEs, the five types of yellow rice wine all had no carcinogenic risks. Under 75% percentile of PAEs concentrations, the DBP in beverage wine with plastic packaging had a carcinogenic risk score of 1.207 5, with a gross carcinogenic risk score of 1.207 5. Under the maximum PAEs concentration, the ross carcinogenic risk scores of cooking wine with plastic packaging, beverage wine with plastic packaging, beverage wine with glass bottle packaging, and beverage wine with jar packaging were 2.751 0, 2.782 0, 1.298 2 and 2.944 0, presenting non-carcinogenic risks. Conclusion There is PAEs contamination in market-available yellow rice wine in Huzhou City, and no carcinogenic risk is evaluated. Non-carcinogenic health risk requires to be given a high priority.

ObjectiveTo explore the therapeutic effect and mechanism of Yiqi Huoxue Tongbian prescription on slow transit constipation （STC） in rats. MethodThe rat model of STC was established by gavage of loperamide hydrochloride. Rats were assigned into control， model， mosapride， low-， medium-， and high-dose （3.51， 7.02， and 14.04 g·kg^-1， respectively） Yiqi Huoxue Tongbian prescription groups. The changes of general signs， fecal moisture content， and intestinal propulsion rate were measured after model establishment and drug administration. The colonic mucosal changes were observed by hematoxylin eosin staining. Enzyme-linked immunosorbent assay was employed to determine the content of substance P （SP） and vasoactive intestinal peptide （VIP） in the colon of rats in each group. The gray values of aquaporin （AQP） 3， AQP4， AQP8， and c-Kit in rat colon tissue were measured by immunohistochemistry and Western blot， and the changes of intestinal flora were detected by 16S rRNA high-throughput sequencing. ResultCompared with the model group， 10 days of treatment with Yiqi Huoxue Tongbian prescription increased the fecal moisture content and intestinal propulsion rate （P<0.01）. The medium- and high-dose Yiqi Huoxue Tongbian prescription groups and the mosapride group showed no obvious mucosal inflammation and neat arrangement of goblet cells with a large number in the colon tissue. Moreover， the three groups showed increased SP content （P<0.01） and decreased VIP content （P<0.01） in the serum. The medium- and high-dose Yiqi Huoxue Tongbian prescription groups showed down-regulated protein levels of AQP3， AQP4， and AQP8 （P<0.01） and up-regulated protein level of c-Kit （P<0.01）. The drug administration groups presented slightly increased observed species， Chao1， ACE， and Shannon， Simpson， and PD whole tree. The principal component analysis showed that the control group had a short distance with the high- and medium-dose Yiqi Huoxue Tongbian prescription groups， indicating that high- and medium-dose Yiqi Huoxue Tongbian prescription can recover the intestinal flora to that in the control group. ConclusionYiqi Huoxue Tongbian prescription can alleviate the defecation status of rats with slow transit constipation by down-regulating the expression of AQP3， AQP4， and AQP8 to reduce the absorption of water in the intestine， up-regulating the expression of c-Kit to increase the number and distribution of Cajal interstitial cells， and regulating the balance of flora in the colon tissue.

Objective:To analyze the molecular mechanism of Zhenqi Fuzheng Capsules in the adjuvant treatment of AIDS by network pharmacology method and molecular docking technology.Methods:The active components and targets of Zhenqi Fuzheng Capsules were obtained through TCMSP, and the AIDS-related targets were obtained through GeneCards, OMIM and DrugBank databases. The intersection target PPI network was constructed through STRING 11.5 database, and Cytoscape 3.9.1 software was used for network topology analysis; Metascape database was used for GO function and KEGG pathway enrichment analysis of core targets; Cytoscape 3.9.1 was used to construct Zhenqi Fuzheng Capsules component-target-pathway network; Autodock Tools software was used to carry out molecular docking of core targets and active components.Results:Totally 31 active components and 180 targets of Zhenqi Fuzheng Capsules were screened out. TNF, IL6, AKT1, IL1B, TP53, VEGFA, RELA, EGFR and CASP3 were identified as the core targets. GO functional enrichment analysis obtained 1 436 biological processes, 53 cellular components, and 117 molecular functions. KEGG pathway enrichment analysis obtained 167 pathways, which were related to pathways in cancer, AGE-RAGE signaling pathway in diabetic complications, and IL-17 signaling pathway was closely related. Molecular docking results showed that core targets such as AKT1 and TNF had good binding activity to quercetin, kaempferol, and luteolin.Conclusion:The main active components of Zhenqi Fuzheng Capsules in the adjuvant treatment of AIDS are quercetin, kaempferol and luteolin, which may treat AIDS through the IL-17 signaling pathway.

Geobacillus thermoglucosidasius is a kind of Gram-positive facultative anaerobic bacteria. The fast growth rate under high temperature and less susceptibility to microbial contamination enable G. thermoglucosidasius to be a desirable producer of biofuels and high-value-added chemicals for the next-generation industrial biotechnology. However, compared with the classical model strain Escherichia coli, the applications of G. thermoglucosidasius are hampered by its low transformation efficiency. This study aimed at obtaining competent cells with high transformation efficiency through inactivating restriction enzymes, adding cell membrane inhibitors and cell wall weakening agents. The results showed that the electro-transformation efficiency achieved 1.2×10⁴ CFU/(μg DNA) by knocking out four genes encoding restriction enzymes. Adding a certain amount of tween 80, dl-threonine and glycine further increased the competent efficiency about 22.5, 44, and 334 times, respectively. The electro-transformation efficiency was enhanced to 4.6×10⁶ CFU/(μg DNA) under the optimized conditions, laying a foundation for genetic manipulation and metabolic engineering of G. thermoglucosidasius.
Electroporation ; Electroporation Therapies ; Bacillaceae ; Cell Membrane ; Escherichia coli/genetics*