OBJECTIVE: To establish a genotyping method for Diego, MNS and Kell blood groups based on quantitative real-time PCR (qPCR) technology, and preliminarily apply it to the screening of rare blood groups in blood donors. METHODS: Blood group gene standards containing heterozygous and homozygous alleles were prepared by blood group serological and PCR-SBT methods. Specific amplification primers and hybridization probes were designed, and explore to establish the qPCR method for detecting Diego, MNS, and Kell blood group genotypes. Then the established qPCR method was used to identify blood group genotypes of 186 blood donor samples. RESULTS: A method based on qPCR technology was established to identify Dia/Dib, S/s and K/k blood group antigens. The genotyping results of the gene standard samples were consistent with the serological testing results and genotypes detected by PCR-SBT. qPCR testing of 186 samples identified 11 cases of DI*A/B heterozygosity and 19 cases of GYPB*S/s heterozygosity, and the rest were DI*B/B, GYPB*s/s, KEL*02/02 homozygosity. No rare blood group genotypes of DI*A/A, GYPB*S/S, KEL*01.01/01.01 were found. CONCLUSION The established qPCR method is suitable for genotyping on Diego, MNS and Kell blood group, and it can be used for batch screening of blood donors and the establishment of rare blood group bank.
Humans ; Genotype ; Genotyping Techniques/methods* ; Real-Time Polymerase Chain Reaction/methods* ; Blood Group Antigens/genetics* ; Kell Blood-Group System/genetics* ; Blood Donors ; Blood Grouping and Crossmatching/methods* ; Erythrocytes ; MNSs Blood-Group System/genetics*

[This corrects the article DOI: 10.1016/j.apsb.2019.01.013.].

Hyssopus cuspidatus is an authentic medicinal herb used in Xinjiang， rich in the chemical constituents including flavonoids， phenolic acids and terpenoids. It possesses anti-inflammatory， antioxidant， and immunomodulatory effects. Modern pharmacological studies have demonstrated that H. cuspidatus exerts therapeutic effects on asthma by inhibiting airway inflammatory responses， suppressing airway remodeling， modulating the function of the hypothalamic-pituitary-adrenal axis， and relaxing bronchial smooth muscle. It also demonstrates intervention effects on chronic obstructive pulmonary disease by reducing levels of inflammatory cytokines and regulating immune balance. Additionally， H. cuspidatus can mitigate acute lung injury by inhibiting oxidative stress and intervene in lung cancer by suppressing proliferation and promoting apoptosis of lung cancer cells. In terms of clinical application， compound preparations containing H. cuspidatus， such as Hanchuan zupa granules and Luo’ou kezupa， have demonstrated favorable therapeutic effects on pulmonary diseases including asthma， cough， and pediatric bronchopneumonia. Currently， the research on precise action targets and compound matching rules of H. cuspidatus remains inadequate. Future studies should integrate modern technologies such as metabolomics to conduct in-depth exploration， thereby promoting the modernized development and clinical application of this traditional medicinal herb.

Apical microsurgery is accurate and minimally invasive, produces few complications, and has a success rate of more than 90%. However, due to the lack of awareness and understanding of apical microsurgery by dental general practitioners and even endodontists, many clinical problems remain to be overcome. The consensus has gathered well-known domestic experts to hold a series of special discussions and reached the consensus. This document specifies the indications, contraindications, preoperative preparations, operational procedures, complication prevention measures, and efficacy evaluation of apical microsurgery and is applicable to dentists who perform apical microsurgery after systematic training.
Microsurgery/standards* ; Humans ; Apicoectomy ; Contraindications, Procedure ; Tooth Apex/diagnostic imaging* ; Postoperative Complications/prevention & control* ; Consensus ; Treatment Outcome

Objective To investigate the distribution and antimicrobial resistance profiles of common pathogens isolated from cerebrospinal fluid(CSF)in CHINET program from 2015 to 2021.Methods The bacterial strains isolated from CSF were identified in accordance with clinical microbiology practice standards.Antimicrobial susceptibility test was conducted using Kirby-Bauer method and automated systems per the unified CHINET protocol.Results A total of 14 014 bacterial strains were isolated from CSF samples from 2015 to 2021,including the strains isolated from inpatients(95.3％)and from outpatient and emergency care patients(4.7％).Overall,19.6％of the isolates were from children and 80.4％were from adults.Gram-positive and Gram-negative bacteria accounted for 68.0％and 32.0％,respectively.Coagulase negative Staphylococcus accounted for 73.0％of the total Gram-positive bacterial isolates.The prevalence of MRSA was 38.2％in children and 45.6％in adults.The prevalence of MRCNS was 67.6％in adults and 69.5％in children.A small number of vancomycin-resistant Enterococcus faecium(2.2％)and linezolid-resistant Enterococcus faecalis(3.1％)were isolated from adult patients.The resistance rates of Escherichia coli and Klebsiella pneumoniae to ceftriaxone were 52.2％and 76.4％in children,70.5％and 63.5％in adults.The prevalence of carbapenem-resistant E.coli and K.pneumoniae(CRKP)was 1.3％and 47.7％in children,6.4％and 47.9％in adults.The prevalence of carbapenem-resistant Acinetobacter baumannii(CRAB)and Pseudomonas aeruginosa(CRPA)was 74.0％and 37.1％in children,81.7％and 39.9％in adults.Conclusions The data derived from antimicrobial resistance surveillance are crucial for clinicians to make evidence-based decisions regarding antibiotic therapy.Attention should be paid to the Gram-negative bacteria,especially CRKP and CRAB in central nervous system(CNS)infections.Ongoing antimicrobial resistance surveillance is helpful for optimizing antibiotic use in CNS infections.

Objective To investigate the antimicrobial resistance of clinical isolates from elderly patients(≥65 years)in major medical institutions across China.Methods Bacterial strains were isolated from elderly patients in 52 hospitals participating in the CHINET Antimicrobial Resistance Surveillance Program during the period from 2015 to 2021.Antimicrobial susceptibility test was carried out by disk diffusion method and automated systems according to the same CHINET protocol.The data were interpreted in accordance with the breakpoints recommended by the Clinical and Laboratory Standards Institute(CLSI)in 2021.Results A total of 514 715 nonduplicate clinical isolates were collected from elderly patients in 52 hospitals from January 1,2015 to December 31,2021.The number of isolates accounted for 34.3％of the total number of clinical isolates from all patients.Overall,21.8％of the 514 715 strains were gram-positive bacteria,and 78.2％were gram-negative bacteria.Majority(90.9％)of the strains were isolated from inpatients.About 42.9％of the strains were isolated from respiratory specimens,and 22.9％were isolated from urine.More than half(60.7％)of the strains were isolated from male patients,and 39.3％isolated from females.About 51.1％of the strains were isolated from patients aged 65-＜75 years.The prevalence of methicillin-resistant strains(MRSA)was 38.8％in 32 190 strains of Staphylococcus aureus.No vancomycin-or linezolid-resistant strains were found.The resistance rate of E.faecalis to most antibiotics was significantly lower than that of Enterococcus faecium,but a few vancomycin-resistant strains(0.2％,1.5％)and linezolid-resistant strains(3.4％,0.3％)were found in E.faecalis and E.faecium.The prevalence of penicillin-susceptible S.pneumoniae(PSSP),penicillin-intermediate S.pneumoniae(PISP),and penicillin-resistant S.pneumoniae(PRSP)was 94.3％,4.0％,and 1.7％in nonmeningitis S.pneumoniae isolates.The resistance rates of Klebsiella spp.(Klebsiella pneumoniae 93.2％)to imipenem and meropenem were 20.9％and 22.3％,respectively.Other Enterobacterales species were highly sensitive to carbapenem antibiotics.Only 1.7％-7.8％of other Enterobacterales strains were resistant to carbapenems.The resistance rates of Acinetobacter spp.(Acinetobacter baumannii 90.6％)to imipenem and meropenem were 68.4％and 70.6％respectively,while 28.5％and 24.3％of P.aeruginosa strains were resistant to imipenem and meropenem,respectively.Conclusions The number of clinical isolates from elderly patients is increasing year by year,especially in the 65-＜75 age group.Respiratory tract isolates were more prevalent in male elderly patients,and urinary tract isolates were more prevalent in female elderly patients.Klebsiella isolates were increasingly resistant to multiple antimicrobial agents,especially carbapenems.Antimicrobial resistance surveillance is helpful for accurate empirical antimicrobial therapy in elderly patients.

Objective To investigate the application value of systemic inflammatory response index(SIRI)in patients with acute-on-chronic liver failure(ACLF)and co-infection.Methods A retrospective analysis was performed for the clinical data of 579 ACLF patients with co-infection who were diagnosed and treated in The Fifth Medical Center of Chinese PLA General Hospital from January 2014 to March 2016,including demographic features,laboratory markers,and complications,and SIRI,Model for End-Stage Liver Disease(MELD)score,MELD combined with serum sodium concentration(MELD-Na)score,and Child-Pugh score were calculated.According to the results of follow-up on day 90,the patients were divided into survival group with 210 patients and death group with 369 patients.The independent-samples t test was used for comparison of normally distributed continuous data between two groups;the Mann-Whitney U rank sum test was used for comparison of non-normally distributed continuous data between two groups;the chi-square test were used for comparison of categorical data between two groups.The binary logistic regression analysis was used to investigate the independent risk factors for 90-day death.The receiver operating characteristic(ROC)curve and the area under the ROC curve(AUC)were used to assess the performance of SIRI,MELD-Na score,and Child-Pugh score in predicting the prognosis of ACLF patients with co-infection.The Kaplan-Meier survival analysis was performed based on the optimal cut-off value of SIRI.Results Among the 597 ACLF patients with co-infection,384(66.32%)had HBV-related ACLF and 114(19.69%)had alcohol-related ACLF;as for the main infection sites,316(54.58%)had abdominal infection and 133(22.97%)had pulmonary infection;the 90-day mortality rate was 63.73%.The multivariate logistic regression analysis showed that SIRI(odds ratio[OR]=1.177,95%confidence interval[CI]:1.117-1.239,P<0.05),blood ammonia(OR=1.009,95%CI:1.001-1.018,P<0.05),MELD-Na score(OR=1.047,95%CI:1.016-1.080,P<0.05),Child-Pugh score(OR=1.351,95%CI:1.054-1.730,P<0.05),age(OR=1.045,95%CI:1.021-1.070,P<0.05),comorbidity with hepatic encephalopathy(OR=2.269,95%CI:1.305-3.946,P<0.05),and comorbidity with acute kidney injury(OR=1.730,95%CI:0.990-3.023,P<0.05)were independent risk factors for 90-day death in ACLF patients with co-infection.The Pearson correlation analysis showed that SIRI was positively correlated with MELD-Na score(r=0.282,P<0.001)and Child-Pugh score(r=0.168,P<0.001).SIRI,MELD-Na score,and Child-Pugh score had an AUC of 0.855,0.734,and 0.690,respectively,in predicting 90-day death,and SIRI had a higher predictive efficiency than MELD-Na score and Child-Pugh score(Z=4.922 and 6.289,both P<0.001),with a sensitivity of 76.7%and a specificity of 82.9%.In addition,SIRI combined with MELD-Na score or Child-Pugh score improved the predictive efficiency of MELD-Na score(0.854 vs 0.734,Z=6.899,P<0.001)and Child-Pugh score(0.858 vs 0.690,Z=8.725,P<0.001).The patients with high SIRI(≥4.08)had a 90-day survival rate of 11.29%(36/319),which was significantly lower than that in the patients with low SIRI(<4.08)(χ2=225.24,P<0.001).Conclusion SIRI is an independent risk factor for death in ACLF patients with co-infection and has a good clinical value in predicting prognosis,with the advantages of convenience and low costs.

OBJECTIVE To establish a preparation system for megakaryocytes(MKs)derived from human embryonic stem cells(hESCs)and MK microparticles(MKMPs),and to assess the pro-angio-genic efficiency of these microparticles.METHODS ①hESCs were induced to mesodermal progenitor cells via monolayer culture with the first-stage induction medium for 2 days before the cells were induced to hemogenic endothelial/hematopoietic progenitor cells by culturing with the second-stage induction medium for another 3 days.Then,the cells were dissociated into single cells,seeded into the third-stage induction medium,and cultured using the suspension method for 8 days to obtain MKs.The specific characters of differentiated cells were identified through morphological observation and flow cytometry before stage-specific marker proteins in different periods were analyzed[hESCs:TRA-1-60,sialyl glycolipid stage-specific embryonic antigen4(SSEA4)];mesodermal progenitor cells:brachyury;hemogenic endothelial/hematopoietic progenitor cells:CD34,CD43;MKs:CD41a,CD42b),and immu-nofluorescence staining[β1-tubulin,von Willebrand factor(VWF)],[friend leukemia integration 1(FLI1),CD42].② MKMP collection and verification:MKMPs were collected via differential centrifugation.The concentration and size of these MKMPs were determined by nanoparticle tracking analysis(NTA),and both the morphology and ultrastructure were examined by transmission electron microscopy(TEM).Besides,the MKMPs-specific proteins[CD41,tumor susceptibility gene 101(TSG101)and CD9]were detected by Western blotting analysis.③ Biological function of MKMPs:MKMPs were stained with CD41a-PE antibodies and co-cultured with human umbilical veinvascular endothelial cells(HUVECs)labeled by CD34-APC for 3 h.Live-cell immunofluorescence was employed to find out whether HUVECs could absorb MKMPs.To find out whether MKMPs could affect the role of HUVECs in angio-genesis and cell migration,platelet microvesicles(PMPs)were used as positive controls.The experi-mental groups were added with different concentrations of microparticles(1,5,10 and 20 mg·L-1)while the control group was given no microparticles(0 mg·L-1).The number of nodes that formed the lumen after 5 h of incubation in Matrigel was counted,and the size of healing of the scratch area was analyzed after 6 h.To elucidate the mechanism through which MKMPs impacted angiogenesis,ELISA was used out to quantitatively detect the concentration of proteins in microparticles.RESULTS ① A three-stage differentiation cultural system was established to develop hESCs into MKs.Flow cytometry revealed progressive loss of pluripotency markers SSEA4 and TRA-1-60,while the mesodermal progenitor marker brachyury peaked at d 2.Subsequently,hemogenic endothelial/hematopoietic progenitor markers CD34 and CD43 emerged at d 5,followed by megakaryocytic markers CD41a and CD42b at d 13.Immunofluorescent images further demonstrated that MKs expressed specific proteins CD42,β1-tubulin,von VWF and FLI1 at d 13.②Microparticles were collected via differential centrifuga-tion.Transmission electron microscopy revealed that their substructure exhibited a typical double-layered membrane.Nanoparticle tracking analysis indicated that the size was(164.3±14.0)nm.The result of WB demonstrated that the microparticles expressed specific markers,including TSG101,CD9 and CD41.③ MKMPs were absorbed after being co-cultured with HUVECs for 3 h and enhanced the ability of HUVECs to form tubes and migrate.Notably,the treatment of 5 mg·L-1 MKMPs was more effective than 5 mg·L-1 PMPs treatment.The results of ELISA showed that the content of VEGF from MKMPs was higher than from PMPs,which may be the key factor in regulating endothelial biological function.CONCLUSION MKs derived from hESCs can generate functional microparticles which can promote angiogenesis.

The classical latent structure method does not consider the influence of primary and secondary symptoms,syndromes and symptoms in the analysis and modeling of syndromes.In this paper,based on the data of damp-heat in intestine and stomach syndrome involving 1087 diabetic patients,the classical latent structure analysis was used to obtain the quantitative syndrome diagnostic rules.Then,using Constrained Latent Tree Analysis(CLTA),the quantitative syndrome diagnostic rules containing primary and secondary symptoms were obtained as follows,primary symptoms include halitosis(2.3),yellow tongue coating(2),abdominal distension(2.3),greasy tongue coating(2.1),loose stool or loose stool(1.5),red tongue(1.3),smooth pulse(1.4).Secondary symptoms include epigastric distension(1.1).Compared with the traditional latent structure analysis method,the rules established by CLTA are more compatible with the concept of differentiating primary and secondary symptoms and the common practice of TCM.The quantitative syndrome diagnostic rules of damp-heat in intestine and stomach syndrome constructed by the CLTA method have considerable objectivity in the modeling process.The diagnostic rules established were also compatible with the qualitative concept of TCM theory in stratifying primary and secondary symptoms.Finally,the diagnostic rules are obtained by logistic regression analysis,and the accuracy of the three rules is compared.The results show that the rule recognition accuracy obtained by CLTA is the highest.Therefore,the syndrome diagnostic rules of damp-heat in intestine and stomach obtained from the analysis of CLTA are in line with the constraint semantics of primary and secondary diseases and the theory of traditional Chinese medicine.

Objective To summarize the changing prevalence of carbapenem resistance in Enterobacterales based on the data of CHINET Antimicrobial Resistance Surveillance Program from 2015 to 2021 for improving antimicrobial treatment in clinical practice.Methods Antimicrobial susceptibility testing was performed using a commercial automated susceptibility testing system according to the unified CHINET protocol.The results were interpreted according to the breakpoints of the Clinical & Laboratory Standards Institute(CLSI)M100 31st ed in 2021.Results Over the seven-year period(2015-2021),the overall prevalence of carbapenem-resistant Enterobacterales(CRE)was 9.43％(62 342/661 235).The prevalence of CRE strains in Klebsiella pneumoniae,Citrobacter freundii,and Enterobacter cloacae was 22.38％,9.73％,and 8.47％,respectively.The prevalence of CRE strains in Escherichia coli was 1.99％.A few CRE strains were also identified in Salmonella and Shigella.The CRE strains were mainly isolated from respiratory specimens(44.23±2.80)％,followed by blood(20.88±3.40)％and urine(18.40±3.45)％.Intensive care units(ICUs)were the major source of the CRE strains(27.43±5.20)％.CRE strains were resistant to all the β-lactam antibiotics tested and most non-β-lactam antimicrobial agents.The CRE strains were relatively susceptible to tigecycline and polymyxins with low resistance rates.Conclusions The prevalence of CRE strains was increasing from 2015 to 2021.CRE strains were highly resistant to most of the antibacterial drugs used in clinical practice.Clinicians should prescribe antimicrobial agents rationally.Hospitals should strengthen antibiotic stewardship in key clinical settings such as ICUs,and take effective infection control measures to curb CRE outbreak and epidemic in hospitals.