AIM To investigate the effects of different compatibility ratios and processing method on the content of rutin,isoquercetin,ferulic acid,quercetin,isotoosendanin,kaempferol,toosendanin,α-pinene,trans-anethole in the combination use of Toosendan Fructus and Foeniculi Fructus,and to explore the optimal compatibility ratio for its use.METHODS The analysis of HPLC-DAD was performed on a 30℃thermostatic ZORBAX SB C18 column(4.6 mm×250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1%phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the use of DAD detector.SPSS 24.0 software was used to analyze the data differences.RESULTS Nine constituents showed good linear relationships within their own ranges(r≥0.999 1),whose average recoveries were 96.19%-103.13%with the RSDs of 1.86%-2.67%.Generally higher total content of nine constituents were detected in the combination use groups when Toosendan Fructus-Foeniculi Fructus were at ratios of 1 ∶ 1,1 ∶ 2,and 2 ∶ 1 than those single uses(P<0.05),and among which the 1 ∶ 1 ratio contributed the highest total content.After salt processing,decreased content of toosendanin and isotoosendanin,α-pinene and trans-anethole(P<0.05,P<0.01)),increased isoquercetin content(P<0.01),and no significant content changes of other ingredients were detected.CONCLUSION Through this method of high accuracy and good reproducibility,we learn that the combination use of Toosendan Fructus and Foeniculi Fructus promotes the dissolution of the nine constituents,and the maximum content is achieved at ratio of 1 ∶ 1.

Objective To investigate the clinical efficacy of mitral valve repair technique in the treatment of rheumatic mitral valve lesions. Methods The clinical data of patients diagnosed with rheumatic mitral valve lesions and undergoing mitral valve repair under extracorporeal circulation in our department from 2021 to 2022 were retrospectively analyzed. Results A total of 100 patients were collected, including 78 females and 22 males with an average age of 52 years. There were no secondary open heart or death in the whole group. Extracorporeal circulation time was 136.3±33.1 min, aortic cross-clamping time was 107.6±27.5 min, ventilator use time was 12.9±5.9 h, ICU stay was 2.6±1.4 d, and vasoactive medication use was 823.4±584.4 mg. Before and after the surgery, there were statistical differences in the left ventricular end diastolic diameter, left atrial end systolic diameter, effective mitral valve orifice area, shortening rate of left ventricular short axis, mitral E-peak blood flow velocity, mean mitral transvalvular pressure difference, mitral pressure half-time, and cardiac function graded by New York Heart Association (P＜0.05). While there was no statistical difference in left ventricular ejection fraction or left ventricular end-diastolic volume (P>0.05). Conclusion Overall repair of rheumatic mitral valve lesions can significantly improve the cardiac function and hemodynamics of the patients, and is a good choice for patients with rheumatic mitral valve lesions.

Objective To explore the efficacy and influencing factors of polyethylene glycol interferon α-2b(Peg-IFNα-2b)combined nucleoside analogues(NAs)in the treatment of hepatitis B virus e-antigen(HBeAg)-negative chronic hepatitis B(CHB)patients who had received NAs treatment,and evaluate the correlation of mononucleotide polymorphisms of interleukin-28B and programmed death receptor-1(PD-1)with interferon treatment response.Methods HBeAg-negative CHB patients who visited Xiangya Hospital of Central South University from January 2020 to December 2022 were analyzed retrospectively.Patients with Peg-IFNα-2b and NAs treatment were as the study group,while those with NAs therapy alone as the control group.Clinical efficacy of two groups of patients at the 12nd,24th,and 48th weeks of treatment,as well as the persistent response and recurrence at the 72nd week were analyzed.PD-1 and IL-28B single nucleotide polymorphisms were adopted to evaluate the value of HBeAg-nega-tive CHB patients in response to interferon treatment.Results At the 48th week of treatment,the response rate of HBeAg-negative CHB patients in the study group was higher than that in the control group(52.05％[38/73]vs 1.64％[1/61],P＜0.05).Among HBeAg-negative CHB patients in the study group,response rates at 48th week of treatment in patients with baseline HBsAg＜100 IU/mL and HBsAg＜1 000 IU/mL were higher than those with HBsAg≥1 000 IU/mL,respectively(both P＜0.05).Univariate and multivariate analyses showed that in HBeAg-negative CHB patients in the study group,the baseline HBsAg levels(OR=1.004,95％CI:1.001-1.006)and HBsAg decline magnitude at the 24th week of treatment(OR=0.111,95％CI:0.034-0.362)were influencing factors for the response of interferon treatment combined with NAs(both P＜0.05).The results of single nucleo-tide polymorphism analysis showed that in HBeAg-negative CHB patients in the study group,the proportion of PD-1 rs10204525 C/T heterozygous mutation in the response population was higher(66.67％vs 16.67％,P＜0.05),while that of IL-28B mutation was not significantly different(P＞0.05).Conclusion Combined treatment with Peg-IFNa-2b can achieve higher HBsAg clearance rate and serological conversion rate in HBeAg-negative CHB patients who had received NAs treatment.HBsAg decline magnitude at the 24th week of treatment can better pre-dict the response at the 48th week of treatment.Patients with low baseline HBsAg level and those carrying PD-1 rs10204525C/T heterozygous mutation gene present better therapeutic effect after receiving Peg-IFNa-2b.

Objective To investigate the effect of fixation of posterior malleolar fractures of Haraguchi type Ⅱ on ankle function in trimalleolar fractures.Methods A total of 118 patients with trimalleolar fractures accompanied with posterior malleolar fractures of Haraguchi type Ⅱ who treated by surgery were selected as the study subjects,and divided into the fixation group(57 cases)and the non-fixation group(61 cases)based on whether the posterior malleolus was fixed during the operation.The operation related indicators,American Orthopaedic Foot and Ankle Society(AOFAS)ankle-hindfoot score,and the excellent and good rate of ankle function recovery were compared between the two groups.Results There was no significant difference in the operation time,intraoperative blood loss,hospital stay,fracture healing time,complete weight-bearing time or postoperative joint flatness between the two groups(P<0.05).At the end of the follow-up,the pain score and total score of the AOFAS ankle-hindfoot scale in the fixation group were higher than those in the non-fixation group,with statistically significant differences(P<0.05);and the excellent and good rate of ankle function recovery in the fixation group was higher than that in the non-fixation group,with statistically significant difference(P<0.05).Conclusion Surgical treatment for the trimalleolar fractures accompanied with posterior malleolar fractures of Haraguchi type Ⅱ can achieve satisfactory clinical outcome,and the fixation of posterior malleolar fractures of Haraguchi type Ⅱ can effectively reduce postoperative joint pain and improve ankle function.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

This study explored a new model of Prostate Imaging Reporting and Data System (PIRADS) and adjusted prostate-specific antigen density of peripheral zone (aPSADPZ) for predicting the occurrence of prostate cancer (PCa) and clinically significant prostate cancer (csPCa). The demographic and clinical characteristics of 853 patients were recorded. Prostate-specific antigen (PSA), PSA density (PSAD), PSAD of peripheral zone (PSADPZ), aPSADPZ, and peripheral zone volume ratio (PZ-ratio) were calculated and subjected to receiver operating characteristic (ROC) curve analysis. The calibration and discrimination abilities of new nomograms were verified with the calibration curve and area under the ROC curve (AUC). The clinical benefits of these models were evaluated by decision curve analysis and clinical impact curves. The AUCs of PSA, PSAD, PSADPZ, aPSADPZ, and PZ-ratio were 0.669, 0.762, 0.659, 0.812, and 0.748 for PCa diagnosis, while 0.713, 0.788, 0.694, 0.828, and 0.735 for csPCa diagnosis, respectively. All nomograms displayed higher net benefit and better overall calibration than the scenarios for predicting the occurrence of PCa or csPCa. The new model significantly improved the diagnostic accuracy of PCa (0.945 vs 0.830, P < 0.01) and csPCa (0.937 vs 0.845, P < 0.01) compared with the base model. In addition, the number of patients with PCa and csPCa predicted by the new model was in good agreement with the actual number of patients with PCa and csPCa in high-risk threshold. This study demonstrates that aPSADPZ has a higher predictive accuracy for PCa diagnosis than the conventional indicators. Combining aPSADPZ with PIRADS can improve PCa diagnosis and avoid unnecessary biopsies.
Male ; Humans ; Prostate/pathology* ; Prostate-Specific Antigen/analysis* ; Prostatic Neoplasms/diagnostic imaging* ; Biopsy ; Nomograms ; Retrospective Studies

Congenital bilateral absence of the vas deferens (CBAVD) is observed in 1%-2% of males presenting with infertility and is clearly associated with cystic fibrosis transmembrane conductance regulator (CFTR) mutations. CFTR is one of the most well-known genes related to male fertility. The frequency of CFTR mutations or impaired CFTR expression is increased in men with nonobstructive azoospermia (NOA). CFTR mutations are highly polymorphic and have established ethnic specificity. Compared with F508Del in Caucasians, the p.G970D mutation is reported to be the most frequent CFTR mutation in Chinese patients with cystic fibrosis. However, whether p.G970D participates in male infertility remains unknown. Herein, a loss-of-function CFTR p.G970D missense mutation was identified in a patient with CBAVD and NOA. Subsequent retrospective analysis of 122 Chinese patients with CBAVD showed that the mutation is a common pathogenic mutation (4.1%, 5/122), excluding polymorphic sites. Furthermore, we generated model cell lines derived from mouse testes harboring the homozygous Cftr p.G965D mutation equivalent to the CFTR variant in patients. The Cftr p.G965D mutation may be lethal in spermatogonial stem cells and spermatogonia and affect the proliferation of spermatocytes and Sertoli cells. In spermatocyte GC-2(spd)ts (GC2) Cftr p.G965D cells, RNA splicing variants were detected and CFTR expression decreased, which may contribute to the phenotypes associated with impaired spermatogenesis. Thus, this study indicated that the CFTR p.G970D missense mutation might be a pathogenic mutation for CBAVD in Chinese males and associated with impaired spermatogenesis by affecting the proliferation of germ cells.
Humans ; Animals ; Mice ; Male ; Mutation, Missense ; Retrospective Studies ; Cystic Fibrosis Transmembrane Conductance Regulator/genetics* ; Infertility, Male/genetics* ; Mutation ; Vas Deferens/abnormalities* ; Spermatogenesis/genetics*

Objective:To evaluate the calf muscle cross-sectional area (CSA) and fat fraction (FF) in amateur marathon runners based on T 2WI-Flex fat-suppression sequence and iterative decomposition of water and fat with echo asymmetrical and least-squares estimation quantitation sequence (IDEAL-IQ) technique, and to explore the correlation between them and age, body mass index (BMI), running age, speed exercise, and monthly exercise. Methods:From September to November 2022, 37 amateur marathon group (marathon group) and 12 healthy volunteers (control group) who did not often exercise were recruited in Shijiazhuang. T 2WI-Flex fat-suppression sequence and IDEAL-IQ sequence were performed on all subjects. The anterior group of the right calf (tibialis anterior, extensor hallucis longus, extensor digital longus), external group of the calf (peroneus longus, peroneus brevis), soleus, medial head and lateral head of the gastrocnemius were delineated along the edge of the muscle contour as regions of interest, and the CSA and FF values of the muscles were obtained. Twelve subjects matched with the clinical data of the control group were selected from the marathon group (marathon matched group), and the differences in CSA and FF were compared by independent sample t test or Mann-Whitney U test. Spearman correlation analysis was used to evaluate the correlation between CSA and FF values of the right calf and running-related indicators including age, BMI, running age, speed exercise, and monthly exercise in 37 amateur marathon runners. Results:The CSA of the right calf soleus muscle in the marathon matched matched group was larger than that in the control group ( t=-2.09, P=0.048). There was no significant difference in CSA of other calf muscles between the two groups ( P>0.05). The FF values of the right calf anterior group, the outer group, the soleus muscle, the medial head and the lateral head of the gastrocnemius muscle in the marathon matched group were lower than those in the control group ( P<0.05). The BMI of 37 amateur marathon runners was positively correlated with the CSA and FF values of the anterior group, the outer group, the soleus, the medial head and the lateral head of the gastrocnemius muscle (CSA: r=0.628, 0.468, 0.680, 0.566, 0.615, respectively, all P<0.05; FF value: r=0.395, 0.567, 0.631, 0.482, 0.516, respectively, all P<0.05). The FF values of the anterior group, the outer group, the soleus, the medial head and the lateral head of the gastrocnemius muscle were negatively correlated with monthly exercise ( r=-0.337, -0.405, -0.437, -0.338, -0.446, respectively, all P<0.05). Conclusions:Long-term running training can reduce the FF value of each calf muscle group, and the FF value of calf muscles is negatively correlated with the amount of running each month. Long-term running training has the greatest effect on the CSA of soleus muscle.

We established an indirect ELISA method using Trichinella spiralis trehalase(TsTRE)protein expressed in prokaryotic cells.The TsTRE gene was amplified by RT-PCR and ligated into the pCold I plasmid,which was expressed in E.coli BL21 competent cells.The rTsTRE protein was purified through affinity column chromatography.The TsTRE protein was localized with immunofluorescence techniques,and the immunogenicity of rTsTRE was detected by westernblotting.Subse-quently,rTsTRE protein was used as a coating antigen to establish an indirect ELISA.We optimized the antigen-coating con-centration,serum dilution concentration,antigen-coating incubation time,type of blocking solution,blocking incubation time,HRP-labeled goat anti-rabbit IgG serum dilution concentration,HRP-labeled goat anti-rabbit IgG serum incubation time and response time of TMB.Subsequently,the critical value,repeatability,sensitivity,specificity and clinical detection rate of the ELISA were evaluated.Immunofluorescence indicated that trehalase was abundant in the rod-shaped body,tail and epidermis of Trichinella spiralis muscle larvae.Western-blot indicated that rTsTRE protein combined with the positive serum of mice infected with T.spiralis for 42 d;the band was approximately 60 kDa.The established indirect ELISA had a positive threshold of 0.384;the intra-run and inter-run coefficients of variation were 5.504％-7.630％ and 4.664％-9.929％,and did not exceed 10％.The lowest detectable titer was 1:1 280.No cross reaction was observed with antibodies to Clonorchissinensis,Schistosoma ja-ponicum,Ascaris suum,Toxocara gondii and Toxocara canis,and the clinical negative detection rate was 0％.Thus,we suc-cessfully expressed the rTsTRE protein.Moreover,the established indirect ELISA method using the TsTRE protein as the coating antigen had good repeatability,sensitivity,specificity and clinical detectability,and can be applied to the detection of clinical samples.

Objective: We analyze the characteristics of Clostridioides difficile (C. difficile) infection among diarrhea patients in Kunming from 2018 to 2020 and provide evidence for follow-up surveillance and prevention. Methods: A total of 388 fecal samples of diarrhea patients from four sentinel hospitals in Yunnan Province from 2018 to 2020 were collected. Real-time quantitative PCR was used to detect the fecal toxin genes of C. difficile. The positive fecal samples isolated the bacteria, and isolates were identified by mass spectrometry. The genomic DNA of the strains was extracted for multi-locus sequence typing (MLST). The fecal toxin, strain isolation, and clinical patient characteristics, including co-infection with other pathogens, were analyzed. Results: Among the 388 fecal samples, 47 samples with positive reference genes of C. difficile were positive, with a total positive rate of 12.11%. There were 4 (8.51%) non-toxigenic and 43 (91.49%) toxigenic ones. A total of 18 strains C. difficile were isolated from 47 positive specimens, and the isolation rate of positive specimens was 38.30%. Among them, 14 strains were positive for tcdA, tcdB, tcdC, tcdR, and tcdE. All 18 strains of C. difficile were negative for binary toxins. The MLST results showed 10 sequence types (ST), including 5 strains of ST37, accounting for 27.78%; 2 strains of ST129, ST3, ST54, and ST2, respectively; and 1 strain of ST35, ST532, ST48, ST27, and ST39, respectively. Fecal toxin gene positive (tcdB+) results were statistically associated with the patient's age group and with or without fever before the visit; positive isolates were only statistically associated with the patient's age group. In addition, some C. difficile patients have co-infection with other diarrhea-related viruses. Conclusions: The infection of C. difficile in diarrhea patients in Kunming is mostly toxigenic strains, and the high diversity of strains was identified using the MLST method. Therefore, the surveillance and prevention of C. difficile should be strengthened.
Humans ; Bacterial Toxins/genetics* ; Enterotoxins/genetics* ; Clostridioides difficile/genetics* ; Multilocus Sequence Typing ; Coinfection ; Bacterial Proteins/genetics* ; China/epidemiology* ; Clostridium Infections/epidemiology* ; Diarrhea/microbiology*