OBJECTIVES: To investigate the effect of in vitro cultured calculus bovis (ICCB) on cerebral ischemia/reperfusion injury (CIRI) and its mechanism. METHODS: A CIRI rat model and a cell model were induced by middle cerebral artery occlusion (MCAO) in Sprague Dawley rats and oxygen glucose deprivation/reperfusion (OGD/R) in BV2 cells, respectively. The CIRI rat model was evaluated using the modified neurological severity score (mNSS), brain water content, and cerebral infarction volume after 1.5 h of ischemia followed by 72 h of reperfusion. Histopathological changes in the cortex and hippocampal CA1 region were observed with hematoxylin and eosin staining. Microglial polarization and NOD-like receptor thermal protein domain associated protein (NLRP) 3 inflammasome expression in the cortex were examined by immunofluorescence. BV2 cell viability was measured via MTT assay after treatment with ICCB and Nigericin. The expressions of NLRP3, ASC, caspase-1 proteins and inflammatory cytokines were detected with Western blotting in OGD/R treated BV2 cells (0.5 h OGD+24 h reperfusion) and in cells pretreated with Nigericin for 24 h. RESULTS: ICCB treatment significantly improved neurological function, reduced cerebral infarct volume and brain water content, and mitigated pathological damage in the cortical and hippocampal CA1 regions of rats subjected to CIRI (all P<0.05). ICCB promoted the transition of cortical microglia from M1 to M2 phenotypes and suppressed NLRP3 activation in microglial cells (all P<0.01). ICCB significantly down-regulated the expression of NLRP3, ASC, and caspase-1 proteins, and reduced the secretion of IL-18 and IL-1β in BV2 cells of OGD/R model (all P<0.01). In addition, Nigericin significantly reversed the salvage effect of ICCB on model cells (both P<0.01) and the modulation of inflammatory cytokines (P<0.05). CONCLUSIONS ICCB exerts a protective effect against CIRI by mitigating neuroinflammation, through the reduction of M1 microglial polarization, promotion of M2 conversion, and suppression of the NLRP3/ASC/caspase-1 signaling pathway.
Animals ; Rats, Sprague-Dawley ; Reperfusion Injury/prevention & control* ; Microglia/metabolism* ; Rats ; NLR Family, Pyrin Domain-Containing 3 Protein ; Brain Ischemia/metabolism* ; Male

This study investigated the expression of the soluble nontoxic mutant CRM197 of diphtheria toxin,to prepare a safe and effective recombinant CRM197 protein.Codon-optimized CRM197 gene sequences were cloned into the pCold Ⅱ and pET-28a(+)prokaryotic expression vectors.The successfully cloned recombinant plasmids were screened and transformed into Escherichia coli BL21(DE3)competent cells.After induction of expression,the protein expression type was determined through SDS-PAGE analysis.The conditions for expression of the solublerecombinant protein were then optimized.The recombinant CRM197 protein was purified through two rounds of Ni-NTA affinity chromatography.The purified protein was used to immunize mice,and antibody levels in the se-rum were measured with ELISA.The codon adaptation index(CAI)of the optimized sequence increased from 0.72 to 0.93.The recom-binant plasmids pET-28a(+)-CRM197 and pCold Ⅱ-CRM197 were successfully constructed,as confirmed through colony PCR and double digestion.Expression analysis revealed that pET-28a(+)-CRM197 was expressed primarily as inclusion bodies,whereas pColdⅡ-CRM197 was expressed predominantly in soluble form.The conditions for soluble protein expression via pCold Ⅱ-CRM197 were optimized.When the inoculum was 3%and the IPTG concentration was 0.4 mmol/L,induction at 20 ℃ for 24 h significantly increased the expression of the soluble target protein.The pCold Ⅱ-CRM197 recombinant protein was purified from the supernatant with Ni-NTA affinity chromatography,thus resulting in a target protein with a purity greater than 98%.ELISA after three rounds of immuniza-tion indicated that the levels of IgG,IgM,IgG1,and IgG2a antibodies in the serum in immunized mice were significantly higher than those in the control group.In summary,the CRM197 recombinant protein was successfully prepared with the pCold Ⅱvector and exhib-ited high soluble expression,high purity,and favorable immunogenicity.

Objectives:To investigate the prognostic value of the CHA2DS2-VASc and R2CHA2DS2-VASc scores in patients with atrial fibrillation(AF)and heart failure(HF).Methods:Patients with AF and HF from hospitals diagnosed by the Heart Failure Center in Guangdong Province between January 2017 and December 2021 were selected.Major adverse cardiovascular events(MACE)were used as the follow-up endpoint.Statistical methods such as the area under the receiver operating characteristic(ROC)curve(AUC),net reclassification index(NRI),and integrated discrimination improvement(IDI)were applied to evaluate the predictive value of the CHA2DS2-VASc and R2CHA2DS2-VASc scores in patients with AF and HF.Results:A total of 1 839 patients were enrolled in this study,comprising 703 patients in the MACE group and 1 136 patients in the non-MACE group.Compared with the non-MACE group,the MACE group exhibited significantly advanced age,higher prevalence of New York Heart Association class Ⅳ and coronary artery disease,lower diastolic blood pressure and estimated glomerular filtration rate levels,and elevated serum N-terminal pro-B-type natriuretic peptide concentrations(all P<0.05).Additionally,significantly lower proportions of patients in the MACE group received angiotensin-converting enzyme inhibitors/angiotensin receptor blockers/angiotensin receptor-neprilysin inhibitors,beta-blockers,mineralocorticoid receptor antagonists,or anticoagulant therapy(all P<0.05).Multivariable logistic regression analysis revealed that each 1-point increment in both CHA2DS2-VASc and R2CHA2DS2-VASc scores was associated with approximately 10%increased risk of MACE.ROC curve analysis demonstrated that the AUC values for predicting MACE in AF patients with HF were 0.555(95%CI:0.528-0.582,P<0.001)for CHA2DS2-VASc and 0.576(95%CI:0.549-0.608,P<0.001)for R2CHA2DS2-VASc,indicating marginally superior discriminatory capacity of the R2CHA2DS2-VASc score.Delong's test confirmed statistically significant differences between the two scoring systems(P=0.001).The R2CHA2DS2-VASc score demonstrated a NRI of 0.259(95%CI:0.166-0.352,P<0.001)and an IDI of 0.007(95%CI:0.005-0.010,P<0.001)compared with the conventional CHA2DS2-VASc score.Although the R2CHA2DS2-VASc score exhibited slightly better predictive accuracy and outcome discrimination capacity than the original scoring system,both scores demonstrated suboptimal clinical predictive performance.Conclusions:Both the R2CHA2DS2-VASc and CHA2DS2-VASc scores show suboptimal performance for predicting the risk of MACE in patients with AF and HF,and the predicting performance of R2CHA2DS2-VASc score is marginally superior to CHA2DS2-VASc score in this patient cohort.

Objectives:To investigate the prognostic value of the CHA2DS2-VASc and R2CHA2DS2-VASc scores in patients with atrial fibrillation(AF)and heart failure(HF).Methods:Patients with AF and HF from hospitals diagnosed by the Heart Failure Center in Guangdong Province between January 2017 and December 2021 were selected.Major adverse cardiovascular events(MACE)were used as the follow-up endpoint.Statistical methods such as the area under the receiver operating characteristic(ROC)curve(AUC),net reclassification index(NRI),and integrated discrimination improvement(IDI)were applied to evaluate the predictive value of the CHA2DS2-VASc and R2CHA2DS2-VASc scores in patients with AF and HF.Results:A total of 1 839 patients were enrolled in this study,comprising 703 patients in the MACE group and 1 136 patients in the non-MACE group.Compared with the non-MACE group,the MACE group exhibited significantly advanced age,higher prevalence of New York Heart Association class Ⅳ and coronary artery disease,lower diastolic blood pressure and estimated glomerular filtration rate levels,and elevated serum N-terminal pro-B-type natriuretic peptide concentrations(all P<0.05).Additionally,significantly lower proportions of patients in the MACE group received angiotensin-converting enzyme inhibitors/angiotensin receptor blockers/angiotensin receptor-neprilysin inhibitors,beta-blockers,mineralocorticoid receptor antagonists,or anticoagulant therapy(all P<0.05).Multivariable logistic regression analysis revealed that each 1-point increment in both CHA2DS2-VASc and R2CHA2DS2-VASc scores was associated with approximately 10%increased risk of MACE.ROC curve analysis demonstrated that the AUC values for predicting MACE in AF patients with HF were 0.555(95%CI:0.528-0.582,P<0.001)for CHA2DS2-VASc and 0.576(95%CI:0.549-0.608,P<0.001)for R2CHA2DS2-VASc,indicating marginally superior discriminatory capacity of the R2CHA2DS2-VASc score.Delong's test confirmed statistically significant differences between the two scoring systems(P=0.001).The R2CHA2DS2-VASc score demonstrated a NRI of 0.259(95%CI:0.166-0.352,P<0.001)and an IDI of 0.007(95%CI:0.005-0.010,P<0.001)compared with the conventional CHA2DS2-VASc score.Although the R2CHA2DS2-VASc score exhibited slightly better predictive accuracy and outcome discrimination capacity than the original scoring system,both scores demonstrated suboptimal clinical predictive performance.Conclusions:Both the R2CHA2DS2-VASc and CHA2DS2-VASc scores show suboptimal performance for predicting the risk of MACE in patients with AF and HF,and the predicting performance of R2CHA2DS2-VASc score is marginally superior to CHA2DS2-VASc score in this patient cohort.

This study investigated the expression of the soluble nontoxic mutant CRM197 of diphtheria toxin,to prepare a safe and effective recombinant CRM197 protein.Codon-optimized CRM197 gene sequences were cloned into the pCold Ⅱ and pET-28a(+)prokaryotic expression vectors.The successfully cloned recombinant plasmids were screened and transformed into Escherichia coli BL21(DE3)competent cells.After induction of expression,the protein expression type was determined through SDS-PAGE analysis.The conditions for expression of the solublerecombinant protein were then optimized.The recombinant CRM197 protein was purified through two rounds of Ni-NTA affinity chromatography.The purified protein was used to immunize mice,and antibody levels in the se-rum were measured with ELISA.The codon adaptation index(CAI)of the optimized sequence increased from 0.72 to 0.93.The recom-binant plasmids pET-28a(+)-CRM197 and pCold Ⅱ-CRM197 were successfully constructed,as confirmed through colony PCR and double digestion.Expression analysis revealed that pET-28a(+)-CRM197 was expressed primarily as inclusion bodies,whereas pColdⅡ-CRM197 was expressed predominantly in soluble form.The conditions for soluble protein expression via pCold Ⅱ-CRM197 were optimized.When the inoculum was 3%and the IPTG concentration was 0.4 mmol/L,induction at 20 ℃ for 24 h significantly increased the expression of the soluble target protein.The pCold Ⅱ-CRM197 recombinant protein was purified from the supernatant with Ni-NTA affinity chromatography,thus resulting in a target protein with a purity greater than 98%.ELISA after three rounds of immuniza-tion indicated that the levels of IgG,IgM,IgG1,and IgG2a antibodies in the serum in immunized mice were significantly higher than those in the control group.In summary,the CRM197 recombinant protein was successfully prepared with the pCold Ⅱvector and exhib-ited high soluble expression,high purity,and favorable immunogenicity.

OBJECTIVE To study on the identification of traditional Chinese medicine of Macleaya cordata(M. cordata) and its similar varieties. METHODS By consulting the ancient herbal books and modern literature, this paper systematically combs and studies the M. cordata. The morphological identification, microscopic identification, physiochemical identification, molecular identification were used to identify M. cordata and its similar varieties. RESULTS Obtained M. cordata herbal textual research data. There were some differences between M. cordata and Macleaya microcarpa(M. microcarpa) and other similar varieties in traits, microscopic, physicochemical and molecular characteristics. Molecular identification results showed that the length of the rbcL gene of M. cordata were 600 bp to 603 bp, with the average GC content ranging from 43.95% to 44.28%. There were significant differences in the variation sites between M. cordata and other similar varieties, and the variation sites with M. microcarpa were the least. The interspecific genetic distance between M. cordata and its similar varieties was greater than its maximum intraspecific genetic distance. NJ analysis results of rbcL could effectively distinguish M. cordata from other similar varieties accurately and quickly. There were significant differences in the secondary structure of rbcL between M. cordata and its similar varieties. CONCLUSION The traditional Chinese medicine identification methods of M. cordata, M. microcarpa and other similar varieties are constructed, which provides experimental basis for the variety identification of M. cordata and the subsequent development of traditional Chinese medicine resources.

OBJECTIVE: To compare the effectiveness of lower extremity axial distractor (LEAD) and traction table assisted closed reduction and intramedullary nail fixation in treatment of femoral subtrochanteric fracture. METHODS: The clinical data of 117 patients with subtrochanteric fracture of femur treated by closed reduction and intramedullary nail fixation between May 2012 and May 2022 who met the selection criteria were retrospectively analyzed. According to the auxiliary reduction tools used during operation, the patients were divided into LEAD group (62 cases with LEAD reduction) and traction table group (55 cases with traction table reduction). There was no significant difference in baseline data, such as gender, age, injured side, cause of injury, fracture Seinsheimer classification, time from injury to operation, and preoperative visual analogue scale (VAS) score, between the two groups ( P>0.05). Total incision length, operation time, intraoperative blood loss, fluoroscopy frequency, closed reduction rate, fracture reduction quality, fracture healing time, weight-bearing activity time, and incidence of complications, as well as hip flexion and extension range of motion (ROM), Harris score, and VAS score at 1 month and 6 months after operation and last follow-up were recorded and compared between the two groups. RESULTS: There were 14 cases in the LEAD group from closed reduction to limited open reduction, and 43 cases in the traction table group. The incisions in the LEAD group healed by first intention, and no complication such as nerve and vascular injury occurred during operation. In the traction table group, 3 cases had perineal crush injury, which recovered spontaneously in 1 week. The total incision length, operation time, intraoperative blood loss, fluoroscopy frequency, and closed reduction rate in the LEAD group were significantly better than those in the traction table group ( P<0.05). There was no significant difference in the quality of fracture reduction between the two groups ( P>0.05). Patients in both groups were followed up 12-44 months, with an average of 15.8 months. In the LEAD group, 1 patient had delayed fracture union at 6 months after operation, 1 patient had nonunion at 3 years after operation, and 1 patient had incision sinus pus flow at 10 months after operation. In the traction table group, there was 1 patient with fracture nonunion at 15 months after operation. X-ray films of the other patients in the two groups showed that the internal fixator was fixed firmly without loosening and the fractures healed. There was no significant difference in fracture healing time, weight bearing activity time, incidence of complications, and postoperative hip flexion and extension ROM, Harris score, and VAS score at different time points between the two groups ( P>0.05). CONCLUSION For femoral subtrochanteric fracture treated by close reduction and intramedullary nail fixation, compared with traction table, LEAD assisted fracture reduction can significantly shorten the operation time, reduce intraoperative blood loss and fluoroscopy frequency, reduce incision length, effectively improve the success rate of closed reduction, and avoid complications related to traction table reduction. It provides a new method for good reduction of femoral subtrochanteric fracture.
Humans ; Fracture Fixation, Intramedullary ; Bone Nails ; Traction ; Blood Loss, Surgical/prevention & control* ; Retrospective Studies ; Treatment Outcome ; Femoral Fractures ; Hip Fractures/surgery* ; Lower Extremity ; Surgical Wound ; Fracture Fixation, Internal

In coronavirus disease 2019 (COVID-19) epidemic prevention and control, effective control of nosocomial infection and prevention of occupational exposure of first-line medical staff are the requirements of scientific prevention and control, as well as the premise and foundation of scientific treatment of patients. This paper introduces the priority of personnel management of infectious occupational exposure risk at The First Affiliated Hospital of Xi’an Jiaotong University, and analyzes engineering control, management and behavior control, and personal protection. It elaborates on the hospital management methods and proposes scientific preventive measures.

OBJECTIVE:To establish a method for simultaneous determination of α-pinene,β-pinene and linalool in volatile oil of Hedychium flavum. METHODS:The volatile oil was extracted from H. flavum according to steam distillation stated in Chinese Pharmacopoeia(2015 edition volume Ⅳ). GC method was adopted to determine the content. The determination was performed on HP-5 capillary chromatographic column(30 m×0.032 mm×0.25 μm)with hydrogen flame ion detector(FID),nitrogen gas as carrier gas,with injector temperature of 200 ℃,and detector temperature of 250 ℃,column flow rate of 0.8 mL/min, samples size of 1 μL and split ratio of 40:1 by programmed temperature. RESULTS:The linear range of α-pinene,β-pinene and linalool were 0.090 5-2.413 3 mg/mL(r=0.999 9),0.098 3-2.620 0 mg/mL(r=0.999 9),0.169 1-4.510 0 mg/mL(r=0.999 8), respectively. RSDs of precision,stability(12 h),reproducibility tests were no more than 2.0%(n=6). The average recoveries were 99.84%(RSD=0.49%,n=6),100.24%(RSD=1.38%,n=6),99.41%(RSD=1.67%,n=6),respectively.The contents of α-pinene,β-pinene and linalool ranged 0.214 4-1.325 0,0.766 2-3.172 1,0.357 4-1.518 7 mg/g in volatile oil from 23 batches of H. flavum produced in Guizhou province. CONCLUSIONS:The method established in this experiment is rapid,simple and accurate.It can be used for the content determination of α-pinene,β-pinene and linalool in volatile oil from H.flavum.