This study was aimed at creating an engineered strain of Bacillus subtilis for efficient expression of biologically active type 2 toxin B(TcdB2)derived from a highly virulent strain of Clostridium difficile.The TcdB2 gene was cloned from ST1/RT027 strain genome DNA,incorporated into the PHT01 vector,and then transformed into B.subtilis strain WB800N for prokaryotic expression.Cell toxicity assays revealed that the recombinant TcdB2 exhibited cytotoxic effects in various cells.The engineered B.subtilis strain effectively expressed biologically active TcdB2,thus providing a basis for further exploration of the pathogenic mechanisms of highly virulent strains of C.difficile and establishing a foundation for potential vaccine can-didate targets.

Objective:To explore the application value of simultaneous monitoring of voriconazole(VRCZ)and voriconazole N-oxide(VNO)in efficacy and safety of VRCZ in the prevention and treatment of fungal infections in allogeneic hematopoietic stem cell transplantation(allo-HSCT)patients before engraftment(i.e.,days+1 to+30 after transplantation).Methods:The influencing factors of VRCZ,VNO concentration and MR(CVNO/CVRCz)and the difference of VRCZ in the prevention and treatment of fungal infection and liver and kidney injury were analyzed.The receiver operating characteristic curve(ROC)was used to analyze the differences(the corresponding to the maximum of the Youden index on the curve was set as the cut-off value)to confirm the critical value.Results:The factors affecting VRCZ concentration(CVRCZ),VNO concentration(CVNO)and MR were patient weight,VRCZ daily dose,and transplantation type(all P＜0.05).CVRCZ and CVNO in the effective group were higher than those in the ineffective group(P＜0.001),the opposite of MR(P＜0.001);the liver and renal injury group had lower MR than the normal group(P＜0.05).ROC showed that CVRCZ,CVNO and MR had important value in predicting VRCZ in the prevention and treatment of invasive fungal infections in allo-HSCT patients before engraftment,and their cutoff of concentrations were 0.95 μg/ml,1.35 μg/ml and 1.645,respectively(AUC:0.9677,0.7634,0.9564).CVRCZ and MR can assist in indicating liver[cutoff values:0.65 μg/ml,1.96(AUC:0.5971,0.6663)]and renal injury[cutoff values:0.95 μg/ml,1.705(AUC:0.6039,0.6164)].Conclusion:The great value of simultaneous monitoring of VRCZ,VNO and MR can predict in the efficacy and safety of VRCZ in allo-HSCT patients before engraftment.The prediction accuracy of CVRCZ was higher than that of MR,followed by that of CVNO.Increased CVRCZ and decreased MR increase the risk of liver and kidney injury.

Objective:To explore the influence of environment temperature on the incidence of testicular torsion.Methods:We collected the clinical data on 172 cases of testicular torsion diagnosed in the Second Hospital of Hebei Medical University from De-cember 2013 to December 2020.According to the local environment temperature on the day of onset,we divided the patients into groups A(below 0℃),B(0-10℃),C(10-20℃)and D(above 20℃),and compared the incidence rates of testicular torsion among the four groups,followed by correlation analysis.Results:The incidence rate of testicular torsion was 12.8％(n=22)in group A,35.5％(n=61)in B,34.9％(n=60)in C and 16.9％(n=29)in D,the highest at 0-10℃ in group B,with sta-tistically significant difference among the four groups(x2=29.07,P＜0.001).Spearman correlation analysis indicated that the inci-dence of testicular torsion was negatively correlated with the environment temperature(r=-0.261,P＜0.01),with no statistically significant difference among different seasons(x2=5.349,P＞0.05),but higher in autumn and winter than in the other two sea-sons.Conclusion:The incidence of testicular torsion is negatively correlated with the environment temperature,elevated when the temperature decreases,but has no statistically significant difference among different seasons,though relatively higher in autumn and winter.

Objective:To investigate the effect of spouse smoking cessation(SSC)on the pregnancy outcome of the patients with polycystic ovary syndrome(PCOS)based on a secondary analysis of the results of a large randomized clinical trial.Methods:Basedon the inclusion and exclusion criteria,this secondary analysis included 861 female participants in the Trial of Acu-puncture Combined with Clomiphene for PCOS from 2011 to 2015.According to the smoking status of their husbands,we divided the subjects into a smoking group(n=345),a non-smoking group(n=387),and a smoking cessation group(n=129),collected their general demographics,anthropometric indexes,laboratory results and pregnancy outcomes,compared them among the three groups,and analyzed the relationship of SSC with the pregnancy outcomes of the PCOS patients.Results:The levels of total testos-terone(T)and low-density lipoprotein cholesterol(LDL-C)in the smoking cessation group were similar to those in the non-smoking group,but significantly lower than those in the smoking group(P=0.014 and P=0.011).The contents of fasting insulin,total cho-lesterol,and apolipoprotein B exhibited statistically significant differences among the three groups(P=0.029,P=0.018 and P=0.024),dramatically lower in the smoking cessation and non-smoking groups than in the smoking group,but similar between the for-mer two.The ovulation rate of the smoking cessation group was similar to that of the non-smoking group,remarkably higher than that of the smoking group(P=0.038).No statistically significant differences were observed in other indicators among the three groups(P＞0.05).Multivariate logistic regression analysis showed that SSC during pregnancy preparation was a protective factor for ovulation in the PCOS patients(OR=1.74,95％CI:1.04-2.92,P=0.035).Conclusion:SSC can improve androgen and glucolipid me-tabolisms of infertile patients with PCOS,and thereby significantly increase the ovulation rate.It is of positive significance to encourage SSC clinically for improving the pregnancy outcomes of infertile women with PCOS.

Objective:To test the expression of miR-146a-5p RNA, interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α) as markers for predicting post-stroke cognitive impairment (PSCI).Methods:Forty cerebral infarction patients who had been followed up after 3 months formed a PSCI group, and another 40 who showed no post-stroke impairment formed the normal (PSCN) group. Forty healthy age-matched people were the AMC group. The executive functioning of each participant was quantified using the digital span test (DST), a Stroop color word test (SCWT), part B of the trail making test (TMT-B), and a semantic fluency test (SFT). Plasma expression levels of miR-146a-5p, IL-6 and TNF-α were also recorded. Receiver operating characteristics (ROC) curves were prepared to analyze the value of the miR-146a-5p, IL-6, TNF-α and Montreal cognitive assessment (MoCA) scores for predicting PSCI.Results:At baseline, the average expression of plasma miR-146a-5p in the PSCI group was significantly lower than in the other groups, with that of the PSCN group significantly higher than the AMC group′s average. Plasma IL-6 content in the PSCI group was significantly higher than in the other two groups on average, with that in the PSCN group significantly higher than in the AMC group. The average TNF-α levels in both the PSCI and PSCN groups were significantly higher than in the AMC group. Three months later, however, the average DST and SFT scores of the PSCI group were significantly lower than those of the other two groups, while TMT-B and stroop interference effects (SIE) times were significantly longer. TMT-B and SIE times in the PSCN group averaged significantly longer than in the AMC group. At baseline, the area under the curve predicting PSCI of plasma miR-146a-5P combined with MoCA scores was 0.90, with a sensitivity of 72.5% and specificity at the optimal critical point of 97.5%.Conclusions:A high level of plasma miR-146a-5p in the acute stage may protect an ACI patient′s cognitive functioning by inhibiting neuroinflammatory responses. Its expression level and the patient′s MoCA score can help to predict PSCI.

Objective:To observe the effects of preventative moxibustion on analgesia,substance P(SP),prostaglandin(PG)F2α and PGE2 in rats with dysmenorrhea due to cold-dampness stagnation,and to explore the analgesic mechanism. Methods:Sixty-four female Wistar non-pregnant rats were randomly divided into a blank group,a model group,a Western medicine group,and a preventative moxibustion group,with 16 rats in each group.Eight qualified diestrus rats were selected from each group.Except for the blank group,the other three groups established models of dysmenorrhea due to cold-dampness stagnation using an ice water bath combined with estradiol benzoate and oxytocin.On the 8th day after modeling,the preventative moxibustion group was treated with gentle moxibustion at Shenque(CV8)and Guanyuan(CV4),and the Western medicine group was given ibuprofen solution for 4 consecutive days.On the 11th day,the intervention groups(i.e.the Western medicine group and the preventative moxibustion group)were treated once again after being injected with oxytocin.The writhing score and the pain threshold of rats were determined;the serum levels of brain-derived neurotrophic factor(BDNF),SP,PGF2α,and PGE2 were measured;the mRNA and protein expression levels of BDNF and its receptor tropomyosin receptor kinase B(TrkB)in the spinal dorsal horn and hypothalamus were detected. Results:Compared with the blank group,the writhing score increased(P<0.01),the pain threshold decreased(P<0.01),the serum levels of BDNF,SP,and PGF2α increased(P<0.01),while the PGE2 decreased(P<0.01);the protein and mRNA expression levels of BDNF and TrkB in the spinal dorsal horn and hypothalamus increased(P<0.01)in the model group.Compared with the model group,the writhing score decreased,the pain threshold increased,the serum BDNF,SP,and PGF2α levels decreased significantly,the serum PGE2 level increased,and the protein and mRNA expression levels of BDNF and TrkB in the spinal dorsal horn and hypothalamus decreased significantly in the preventative moxibustion group and the Western medicine group,while the inter-group differences were significant(P<0.01).Compared with the Western medicine group,the writhing score decreased,the pain threshold increased,the serum BDNF,SP,and PGF2α,levels decreased,the serum PGE2 level increased,and the protein and mRNA expression levels of BDNF and TrkB in the spinal dorsal horn and hypothalamus in the preventative moxibustion group decreased significantly,while the inter-group differences were significant(P<0.05 or P<0.01). Conclusion:Preventative moxibustion at Shenque(CV8)and Guanyuan(CV4)can improve the pain sensitization state of rats with dysmenorrhea due to cold-dampness stagnation,down-regulate the mRNA and protein expression levels of BDNF and TrkB in the spinal dorsal horn and hypothalamus;regulation of the serum SP,PGF2α,and PGE2 levels may be part of the mechanism.

Epigenomic imbalance drives abnormal transcriptional processes,promoting the onset and progression of cancer.Although defective gene regulation generally affects carcinogenesis and tumor suppression networks,tumor immunogenicity and immune cells involved in antitumor responses may also be affected by epigenomic changes,which may have significant implications for the development and application of epigenetic therapy,cancer immunotherapy,and their combinations.Herein,we focus on the impact of epigenetic regulation on tumor immune cell function and the role of key abnormal epigenetic processes,DNA methylation,histone post-translational modification,and chromatin structure in tumor immunogenicity,and introduce these epigenetic research methods.We emphasize the value of small-molecule inhibitors of epigenetic modulators in enhancing antitumor immune responses and discuss the challenges of developing treatment plans that combine epigenetic therapy and immuno-therapy through the complex interaction between cancer epigenetics and cancer immunology.

Aim To investigate the role of metabolites of eicosapentaenoic acid (EPA) in promoting the transdifferentiation of pancreatic α cells to β cells. Methods Male C57BL/6J mice were injected intraperitoneally with 60 mg/kg streptozocin (STZ) for five consecutive days to establish a type 1 diabetes (T1DM) mouse model. After two weeks, they were randomly divided into model groups and 97% EPA diet intervention group, 75% fish oil (50% EPA +25% DHA) diet intervention group, and random blood glucose was detected every week; after the model expired, the regeneration of pancreatic β cells in mouse pancreas was observed by immunofluorescence staining. The islets of mice (obtained by crossing GCG

Objective:To explore the feasibility of using self-made visual throat forceps to remove hypopharyngeal foreign bodies. Methods:The throat forceps were combined with the endoscope and connected to a monitor via a data cable resulting in a visual throat forceps apparatus. This device was utilized to examine and treat the hypopharyngeal foreign bodies. Results:Among 53 patients, foreign bodies were detected in 51,with 48 cases involving hypopharyngeal foreign bodies. All were successfully extracted using the visual throat forceps. Three cases, diagnosed as esophageal foreign bodies by electronic gastroscopy, were treated using the same method. Conclusion:Visual throat forceps can be used to examine the hypopharynx and remove foreign bodies. It has the advantages of simple operation, rapid operation, and high success rate of foreign body removal from the hypopharynx. It is worthy of clinical application.
Humans ; Hypopharynx/surgery* ; Pharynx/surgery* ; Endoscopes ; Surgical Instruments ; Foreign Bodies/diagnosis*

Decoction is the most commonly used dosage form in the clinical treatment of traditional Chinese medicine (TCM). During boiling, the violent movement of various active ingredients in TCM creates molecular forces such as hydrogen bonding, π-π stacking, hydrophobic interactions and electrostatic interactions, which results in the formation of self-assembled aggregates in decoction (SADs), including particles, gels, fibers, etc. It was found that SADs widely existed in decoction with biological activities superior to both effective monomers and their physical mixtures, providing a new idea to reveal the pharmacodynamic material basis of Chinese herbal medicine from the perspective of component interactions-phase structure. Recently, SADs have become a novel focus of research in TCM. This paper reviewed their relevant studies in recent years and found some issues to be concerned in the research, such as the polydispersity of decoction system, instability of active ingredient interactions during boiling, uncertainty of the aggregates self-assembly rules, and stability, purity, yield of the products. In this regard, some solutions and new ideas were presented for the integrated development and clinical application of SADs.