Intervertebral disc degeneration (IDD) is largely attributed to impaired endogenous repair. Nucleus pulposus-derived stem cells (NPSCs) senescence leads to endogenous repair failure. Small extracellular vesicles/exosomes derived from mesenchymal stem cells (mExo) have shown great therapeutic potential in IDD, while whether mExo could alleviate NPSCs senescence and its mechanisms remained unknown. We established a compression-induced NPSCs senescence model and rat IDD models to evaluate the therapeutic efficiency of mExo and investigate the mechanisms. We found that mExo significantly alleviated NPSCs senescence and promoted disc regeneration while knocking down thioredoxin (TXN) impaired the protective effects of mExo. TXN was bound to various endosomal sorting complex required for transport (ESCRT) proteins. Autocrine motility factor receptor (AMFR) mediated TXN K63 ubiquitination to promote the binding of TXN on ESCRT proteins and sorting of TXN into mExo. Knocking down exosomal TXN inhibited the transcriptional activity of nuclear factor erythroid 2-related factor 2 (NRF2) and activator protein 1 (AP-1). NRF2 and AP-1 inhibition reduced endogenous TXN production that was promoted by exosomal TXN. Inhibition of NRF2 in vivo diminished the anti-senescence and regenerative effects of mExo. Conclusively, AMFR-mediated TXN ubiquitination promoted the sorting of TXN into mExo, allowing exosomal TXN to promote endogenous TXN production in NPSCs via TXN/NRF2/AP-1 feed-forward circuit to alleviate NPSCs senescence and disc degeneration.

Objective:To investigate the changes of mitochondria in the substantia nigra pars reticulata(SNr)in a mouse model of acute hepatic encephalopathy(AHE),and the effects of mitochondrial division inhibitor Mdivi-1 on the motor function and mitochondrial function of SNr in AHE mice.Methods:The mouse model of AHE was established by intraperitoneal injection of thioacetamide(TAA)and treated with Mdivi-1.The changes of serum aspartate aminotrans-ferase(AST),alanine aminotransferase(ALT),and blood ammonia were detected by biochemical detection kits.Open field test,rotor-rod fatigue test and elevated plus maze test were performed to observe the motor function of AHE mice.Mitochondrial membrane potential(MMP),cellular reactive oxygen species(ROS)and ATP of SNr were detected by commercial kits.Results:Compared with the control group,the levels of AST,ALT and blood ammonia in AHE mice were increased.The total movement distance of the mice in the open field was reduced,and the movement time of the rotor-rod fatigue test and the elevated plus maze test were shortened.In SNr,mitochondria became smaller and rounder,mitochondrial fission increased,MMP decreased,cellular ROS increased,and ATP production decreased.After treat-ment with Mdivi-1,the levels of AST,ALT and blood ammonia in AHE mice were decreased.In the open field,the total movement distance of mice increased,the movement time of rotorrod fatigue test and elevated plus maze test increased,the mitochondria of SNr were larger,with decreased roundness,decreased mitochondrial division,increased MMP,decreased cellular ROS,and increased ATP production.Conclusion:Mdivi-1 can improve movement disorders in AHE mice by repairing mitochondrial in the SNr.

Objective:To investigate effect of neferine(NeF)on myocarditis in heart failure(HF)rats by regulating CC-che-mokine ligand 2/CC chemokine receptor 2(CCL2-CCR2)signaling axis.Methods:HF rat model was constructed by ligating left ante-rior descending branch,and randomly separated into HF group,different doses of NeF groups(5 mg/kg,10 mg/kg,20 mg/kg),20 mg/kg NeF+CCL2 group,another 10 rats without ligating left anterior descending branch were regarded as sham surgery group.After treatment,cardiac function[left ventricular end systolic diameter(LVESD),ejection fraction(EF),left ventricular ejection fraction(LVEF)]were evaluated.Serum indicators[brain natriuretic peptide(BNP),IL-1β,N-terminal B-type natriuretic peptide(NT-pro BNP),TNF-α]were detected.Myocardial tissue was isolated and expressions of TNF-α and IL-1β,pathological damage,collagen deposition,expressions of CCL2 and CCR2 proteins were detected.Results:Compared with sham surgery group,HF group showed a decrease in myocardial cells,infiltration of inflammatory cells,and severe deposition of collagen fibers,LVESD,serum BNP,NT-pro BNP,TNF-α,IL-1β levels,myocardial tissue TNF-α,IL-1β expressions,collagen area ratio,CCL2,CCR2 protein expressions were greatly increased,LVEF and EF were greatly decreased(P<0.05).Compared with HF group,central muscle tissue damage and collagen fiber deposition were reduced in different doses of NeF groups,LVESD,serum BNP,NT-pro BNP,TNF-α,IL-1β levels,myocardial tissue TNF-α,IL-1β expressions,collagen area ratio,CCL2,CCR2 protein expressions were greatly decreased,LVEF and EF were greatly increased,with differences between groups(P<0.05).After CCL2 intervention,compared with 20 mg/kg NeF group,myocardial tissue damage and collagen fiber deposition were worsened,LVESD,serum BNP,NT-pro BNP,TNF-α,IL-1β levels,myocardial tissue TNF-α,IL-1β expressions,collagen area ratio,CCL2,CCR2 protein expressions were greatly increased,LVEF and EF were greatly decreased(P<0.05).Conclusion:NeF regulates CCL2-CCR2 signaling axis,inhibits myocar-ditis in HF rats,and improves cardiac function.

To assess the positive predictive value (PPV) of extended noninvasive prenatal testing (NIPT-plus) for fetal chromosomal abnormalities. This retrospective research enrolled 511 cases of pregnant women with positive NIPT-plus results at the Obstetrics and Gynecology Hospital of Fudan University from May 2017 to January 2021. Karyotype analysis and chromosome microarray analysis (CMA) techniques was applied for verification. All cases were followed to determine their pregnancy outcome. The Chi-square test was used in PPV. 63 out of 511 refused prenatal diagnosis after counseling, 448 pregnant women with prenatal diagnosis showed that the PPVs of NIPT-plus test for fetal trisomy 21, 18 and 13 (T21, T18, T13), sex chromosome aneuploidy (SCAs) and chromosome microdeletion/microduplication syndrome (MMS) were 86.0% (92/107), 79.5% (35/44), 54.5% (12/22), 39.5% (75/190), and 41.7% (30/72), respectively. The results revealed that the PPV was higher among older pregnant women compared to young pregnant women (77.8% vs. 51.9%, P<0.01). With increasing maternal age, the PPV of NIPT-plus presented increasing trends for T21, T13, and composite PPV except for T18 or SCAs. In addition, the termination rates for confirmed SCAs fetal karyotypes 45, X; 47, XXX; 47, XXY and 47, XYY were 11/11, 3/15, 91.7% (22/24) and 1/14, respectively. NIPT-plus can safely and effectively detect fetal chromosomal abnormalities and can be extended to MMS screening, significantly reducing the proportion of interventional prenatal diagnoses, and those with positive screening still require further confirmation.

To assess the positive predictive value (PPV) of extended noninvasive prenatal testing (NIPT-plus) for fetal chromosomal abnormalities. This retrospective research enrolled 511 cases of pregnant women with positive NIPT-plus results at the Obstetrics and Gynecology Hospital of Fudan University from May 2017 to January 2021. Karyotype analysis and chromosome microarray analysis (CMA) techniques was applied for verification. All cases were followed to determine their pregnancy outcome. The Chi-square test was used in PPV. 63 out of 511 refused prenatal diagnosis after counseling, 448 pregnant women with prenatal diagnosis showed that the PPVs of NIPT-plus test for fetal trisomy 21, 18 and 13 (T21, T18, T13), sex chromosome aneuploidy (SCAs) and chromosome microdeletion/microduplication syndrome (MMS) were 86.0% (92/107), 79.5% (35/44), 54.5% (12/22), 39.5% (75/190), and 41.7% (30/72), respectively. The results revealed that the PPV was higher among older pregnant women compared to young pregnant women (77.8% vs. 51.9%, P<0.01). With increasing maternal age, the PPV of NIPT-plus presented increasing trends for T21, T13, and composite PPV except for T18 or SCAs. In addition, the termination rates for confirmed SCAs fetal karyotypes 45, X; 47, XXX; 47, XXY and 47, XYY were 11/11, 3/15, 91.7% (22/24) and 1/14, respectively. NIPT-plus can safely and effectively detect fetal chromosomal abnormalities and can be extended to MMS screening, significantly reducing the proportion of interventional prenatal diagnoses, and those with positive screening still require further confirmation.

Objective:To investigate the clinical effect of hysteroscopic scar excision for cesarean scar pregnancy(CSP) and its effect on serum β-human chorionic gonadotropin(β-HCG).Methods:From January 2017 to January 2019, 83 patients with endogenous CSP admitted to the People's Hospital of Wenzhou were divided into observation group(43 cases) and control group(40 cases) according to different surgical methods.The observation group underwent hysteroscopic scar lesion resection, while the control group underwent uterine artery embolization.The success rate, intraoperative bleeding volume, vaginal bleeding time and hospitalization time, postoperative complications and the recovery time of blood β-HCG were compared between the two groups.Results:The success rate of operation in the observation group was 97.67%(42/43), which was higher than that in the control group[82.50%(33/40)](χ 2=3.875, P<0.05). The amount of bleeding in the observation group[(43.28±10.28)mL] was less than that in the control group[(74.32±15.48)mL], the time of vaginal bleeding in the observation group[(5.79±1.45)d] was shorter than that in the control group[(8.54±1.89)d], and the time of hospitalization in the observation group[(7.32±1.48)d]was shorter than that in the control group[(10.80±1.97)d], the differences were statistically significant( t=10.832, 7.468, 9.140, all P<0.05). There was no statistically significant difference in postoperative complications between the two groups( P>0.05). The recovery time of blood β-HCG in the observation group[(19.54±3.52)d]was shorter than that in the control group[(28.73±6.57)d]( t=8.020, P<0.05). The total satisfaction rate of the observation group was 97.67%(42/43), which was higher than 77.50%(31/40) of the control group(χ 2=6.170, P<0.05). Conclusion:Hysteroscopy-assisted scar excision is effective in the treatment of CSP, and can shorten the recovery time of blood β-HCG.

Objective To investigate the prevalence and cardiovascular risk factors of silent brain infarct (SBI) in Shunyi Cohort.Methods This study was based on the population based Shunyi Study in China.One thousand and twenty-seven stroke-free participants older than 35 years,who completed cerebral MRI,were included.Cardiovascular risk factors were assessed by interview,physical examination and blood sample tests.SBI was evaluated on 3D-T1WI,T2WI and FLAIR sequences.Associations between risk factors and SBI were analyzed by Logistic regression and adjusted for age,sex,and relevant confounders.Results One thousand and twenty-seven participants,aged (55.9 ± 9.4) years,37.7％ male,were assessed.One hundred sixty-four participants(16.0％) had SBI on MRI.The prevalence of SBI increased by age (every 10 years,OR=2.12,95％ CI 1.74-2.58,P＜0.01).Hypertension(OR =2.67,95％ CI 1.77-4.04,P＜0.01),diabetes(OR=2.48,95％ CI 1.64-3.76,P＜0.01) and smoking(OR=1.98,95％ CI 1.08-3.62,P =0.028) were significantly associated with SBI.Conclusions The prevalence of SBI in this Chinese population is 16.0％,which increases with age.Hypertension,diabetes and smoking are associated with SBI.

Objective Toinvestigateantimicrobialresistanceamonggram-positivecocciinChinain 2013.Methods Retrospectivestudy.FromJune2013toDecember2013,1663consecutiveandnon-repetitive gram-positive cocci were collected from 15 teaching hospitals. The minimal inhibitory concentration ( MIC) of antibacterial agents was determined by agar dilution method. A retrospective study was conducted on rates of resistance to antimicrobial agents. The prevalence of penicillin-resistant Streptococcus pneumoniae ( PRSP) between children and adult patients and the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) between elder group and younger adult patients were compared using chi-square test. Results The prevalence of PRSP in children below 3 years old ( 72. 9%, 51/70 ) was higher than adult patients (55. 2%, 106/192) (χ2 =6. 653,P<0. 05). About 94. 9%(261/275) and (92. 7%,255/275) of S. pneumonia were resistant to erythromycin and clindamycin. All S. pneumoniae strains were susceptible to teicoplanin, vancomycin, linezolid, tigecycline and daptomycin. Penicillin still showed very high activity against Streptococcus spp. β-Hemolytic group. More than 60% of Streptococcus spp.β-Hemolytic group were resistant to erythromycin, clindamycin and tetracyclines. The prevalence of MRSA and methicillin-resistant coagulase-negative Staphylococci(MRCoNS) was 39. 7%(229/576) and 80. 6%(224/278), respectively. The MRSA prevalence ranged from 24. 2% to 70. 0% in different regions. About 52. 6%( 100/190 ) of Staphylococcus aureus from respiratory tract specimens, 38. 5%(40/104)of Staphylococcus aureus from blood samples, and 29. 7%(58/195) of Staphylococcus aureus from wound and pus were resistant to methicillin. The prevalence of MRSA in elder group ( 48. 6%, 84/173 ) was higher than that in younger adult patients (35. 7%, 144/403)(χ2 =8. 322,P <0. 05). The susceptibility rates of MRSA to chloramphenicol and trimethoprim/sulfamethoxazole were 86. 4% ( 244/228 ) and 94. 7% ( 237/228 ) , respectively. Susceptibility rates to gentamycin, erythromycin, clindamycin, tetracyclines, rifampicin and quinolones were ranged from 15. 8% to 59. 6%. All Staphylococci isolates were susceptible to teicoplanin, vancomycin, linezolid, daptomycin and tigecycline. All Enterococcus isolates were susceptible to daptomycin and tigecycline. All E. faecalis ( 158/158 ) and 96. 4% ( 133/138 ) of E. faecium were susceptible to teicoplanin. About 98. 0% ( 150/153 ) of E. faecalis and 97. 1% ( 145/138 ) of E. faecium were susceptible to linezoild. About 45. 8% (70/153) of E. faecalis and 60. 9% (84/138) of E. faecium were resistant to gentamycin with a high concentration. The susceptibility of E. faecalis to all the antibiotics tested exceptchloramphenicolandtetracyclinewashigherthanthatofE.faecium.Conclusions Basedon different age groups and regions, the resistance rates of Gram-positive cocci are different. Teicoplanin, vancomycin, tigecycline, daptomycin, linezolid and tedizolid showed very high activity against Gram-positive cocci. (Chin J Lab Med,2015,38:373-381)

Objective To evaluate the relationship between functional oipA gene of Helicobacterpylori and digestive disease. Methods Biopsy gastric mucosa were obtained from 360 patients who would get gastroscopy.Hp were isolated from urease positive samples and partly urease negative samples,and improved by microscope,urease experiment and 16SrRNA PCR. OipA and cagA of the isolates were obtained by PCR and the statas of oipA signal region were analysised after sequencing. Analyze the relevance between functional oipA gene of Helicobacter pylori and digestive disease.Results 106 isolated Hp were obtained,in which 72 strains with cagA gene positive,87 with oipA signal region gene positive (80 the statas of signal region was on and 1 was off;6 could not be decided).The frequency of functional oipA were both significantly higher than cagA in ulcer and atrophic gastritis.Conclusion The relationship between functional oipA gene and digestive disease was more closer than that of cagA.

Objective To establish an indirect ELISA method with Helicobacter pylori (HP)recombinated antigene OipA6 as coating antigen after prokaryotic expression and purification.Methods The ELISA plate was coated with Hp OipA6 recom-binant protein as antigen.Serum specimens and enzyme labeled anti human IgG were added.The reaction conditions was op-timized.The results of indirect ELISA were compared with the detection of oipA gene.Results When Hp OipA6 recombi-nant protein was 1.0 mg/ml,the condition of coating was 2~8℃ one night,the dilution of serum specimens was 1∶100 and enzyme labeled anti human IgG was 1∶4 000,the incubation time of enzyme labeled anti human IgG was 40 min,the chro-mogenic time was 15 min,the result of ELISA was the best.The sensitivity and specificity of the indirect ELISA were both higher than the detection of oipA gene.Conclusion It suggested that the indirect ELISA,in which Hp OipA6 recombinant protein were coated,may be an appropriate assisted method in detection of high virulent Hp infection.