Objective To exPlore the effect of mammalian target of raPamycin ( mTOR ) inhibitor eVerolimus on radiosensitiVity of human non_small cell lung cancer cell line in vitro by using eVerolimus to inhibit mTOR signaling Pathway of A549. Methods Human non_small cell lung cancer cell line A549 was subjected to radiation alone or in combination with eVerolimus treatment. The 50%inhibition concentration ( IC50 ) of eVerolimus in A549 cells was detected by methylthiazol tetrazolium ( MTT) assay in vitro. EVerolimus at the 20%inhibition concentration ( IC20 ) was used to Pretreat A549 cells for 24 h. Cells were then irradiated by X_ray with 2,4,6,8 Gy. The cell surViVal fraction was comPuted by clone formation. Cell surViVal curVe was fitted by multitarget one_hit model, and mean lethal dose ( D0 ), dose quasithreshold ( Dq ), surViVal fraction at 2 Gy ( SF2 ), and sensitization enhancement ratio (SER) were calculated. The exPression ofγ_H2AX was determined by Western blotting and then the relatiVe gray Values were analyzed. Results EVerolimus significantly imProVed the sensitiVity of A549 cells to radiation. The D0 , Dq and SF2 of eVerolimus+irradiation grouP were significantly lower than those of irradiation grouP. The SER was 1. 36. The residual amount of γ_H2AX Protein in the eVerolimus + irradiation grouP was significantly higher than that of the irradiation grouP. Conclusion EVerolimus inhibiting mTOR signaling Pathway can increase the radiosensitiVity of A549 cells.

Objective To investigate the effect of acupunctures variable with stages on lumbar intervertebral dise protrusion (LIDP). Methods 98 LIDP patients in our hospital from October, 2014 to July, 2015 were randomly divided into control group (n=49) and observa-tion group (n=49) according to the sequence of first diagnosis time. The control group received routine acupuncture, and the observation group acupunctured at Ouch point and Huantiao point (GB30) with different acupuncture according to manifestations and courses. Modified Yang acupuncture was adopted in acute phase, triple acupuncture was used in remittent phase, and lateral needling was used in the recovery phase. They were both treated once a day, 7 days as a course for 3 courses with 2 days of interval. Visual Analogue Scale (VAS), Oswestry Disability Index (ODI) and clinical effects were assessed before and 1, 2, 3 courses after treatment. Results Both VAS and ODI scores im-proved after treatment (F>7.12, P<0.05), especially in the observation group (t>4.43, P<0.05). The efficiency was higher in the observation group than in the control group (χ2=5.594, P<0.05). Conclusion Compared with normal acupuncture, the acupunctures variable with stages is more effective on LIDP than routine.

Objective: To construct a prediction model for preeclampsia (PE) risk in twin-pregnant women, so as to provide the basis for early screening and prevention of PE. Methods: A total of 467 twin-pregnant women who underwent prenatal examination and delivered at Huzhou Maternal and Child Health Hospital were selected. Sixty cases with preeclampsia (PE) were included in the case group, and 60 women without PE were included in the control group. General information, blood biochemical indicators and uterine artery resistance index (UtA-RI) were collected. A logistic regression model was used to screen predictive factors and establish a nomogram. The Bootstrap method was performed for the internal validation; the receiver operating characteristic (ROC) curve, calibration curve and decision curve analysis were employed to evaluate the discrimination, calibration and clinical utility of the nomogram, respectively. Results: In the case group, there were 47 individuals (78.33%) aged younger than 35 years, 21 individuals (35.00%) with pre-pregnancy body mass index (BMI) of 25 kg/m2 and above, and 33 individuals (55.00%) with in vitro fertilization. In the control group, there were 57 individuals (95.00%) aged younger than 35 years, 8 individuals (13.33%) with pre-pregnancy BMI of 25 kg/m2 and above, and 39 individuals (65.00%) with natural pregnancy. Multivariable logistic regression analysis identified age, pre-pregnancy BMI, method of conception, placental growth factor (PLGF) and UtA-RI as risk prediction factors for PE in twin-pregnant women. The established nomogram had an area under the ROC curve of 0.827 (95%CI: 0.755-0.899), a sensitivity of 0.767, a specificity of 0.733, a good discrimination and calibration, and a relatively high clinical net benefit. Conclusion The nomogram established by age, pre-pregnancy BMI, method of conception, PLGF and UtA-RI has a good predictive value for the risk of PE in twin-pregnant women.

Objective:To investigate the detection rate of intestinal colonization of carbapenem-resistant Enterobacterales (CRE) in inpatients, and to analyze the molecular epidemiological characteristics of CRE strains.Methods:This was a prospective study. Stool, rectal swab or perianal swab specimens of 213 inpatients in the surgical intensive care unit (SICU), medical intensive care unit (MICU) and the department of hematology (transplantation ward) in The Second Hospital of Anhui Medical University were collected from March to December, 2019. MacConkey plate containing carbapenems was used to screen CRE strains, and bacteria identification and drug susceptibility test were conducted. Key strains were selected for whole genome sequencing (WGS). Besides, multilocus sequence typing, capsular serotype, drug resistance gene, virulence gene and plasmid carrying characteristics of these strains were analyzed. Using KPN FJ723042 sequence as a reference, the single-nucleotide polymorphism (SNP) of all strains was analyzed.Results:Twenty-three CRE strains were detected, with a detection rate of 10.8%(23/213), which included 15(65.2%) carbapenem-resistant Klebsiella pneumoniae (CRKP) isolates, three (13.0%) Escherichia coli strains, three (13.0%) Enterobacter cloacae strains and two (8.7%) Citrobacter freundii strains. SNP cluster analysis showed that the 15 CRKP strains had two main clonotypes, which were both predominant in SICU. Fifteen strains of CRKP were type ST11-K64. All these strains carried β-lactamase Klebsiella pneumoniae carbapenemase 2 ( blaKPC-2 ), and 12 strains carried regulator of mucoid phenotype gene A2 ( rmpA2) and iucABCD. Conclusions:The detection rate of intestinal colonization of CRE in inpatients is high, and most strains are CRKP of type ST11-K64. These CRKP strains have both multidrug resistance and virulence characteristics, which are risks for hospital transmission.

Objective:The whole-genome sequencing and virulence characteristics analysis of a Klebsiella pneumoniae isolate that caused lower limb gas gangrene were performed to provide a reference for the comprehensive understanding of molecular virulence characteristics of K. pneumoniae causing severe community-acquired infection. Methods:The patient was admitted to the emergency department for treatment on March 13, 2018.The main clinical symptoms of the patients were high fever, gas gangrene of the left lower limb, and diabetic ketoacidosis. The pus specimen was collected for the bacterial culture, isolates identification and antimicrobial susceptibility testing. Hypermucoviscous phenotype was detected by string test. The whole genome of the isolate was sequenced and the multi-site sequence typing, capsular serotyping, plasmid characteristics, virulence and antimicrobial resistance genes of the isolate were analyzed. Plasmid curing and conjugation experiments were used to analyze plasmid characteristics. The virulence of the strain was assessed by serum killing and Galleria mellonella lethality assays. A two-sample t-test was used to compare the differences in the lethal dose of 50% (LD 50) between the tested strains and reference strains against the G. mellonella larvae. Results:K. pneumoniae strain KPN41053 was identified, it was only resistant to ampicillin and was negative for hypermucoviscous phenotype. Whole genome sequencing showed that the length of KPN41053 chromosome was 5 377 071 bp, belonging to ST660 type, and the capsular type was K16. A IncFIB(K)/HI1B virulence plasmid (207 506 bp) with a sequence that was highly similar to pLVPK was harbored by KPN41053. The plasmid carried a variety of virulence genes, among which rmpA and rmpA2 were pseudogenes. The plasmid could not be transferred horizontally by conjugation. The variation strain KPN41053_PC was obtained by plasmid curing. Serum killing analysis showed that KPN41053 was serum resistant (Grade 6), and KPN41053_PC was serum intermediately sensitive (Grade 3). The lgLD 50 of KPN41053 had no difference with that of the hypervirulent control strain (ST23-K1 type) ( t=0.32, P=0.765), and was significantly lower than that of KPN41053_PC ( t=5.97, P=0.004). Conclusions:KPN41053 was an atypical hypervirulent K. pneumoniae that belonged to ST660 but without a hypermucoviscous phenotype. The virulence plasmid harbored by KPN41053 was its key virulence factor. Hypervirulent K. pneumoniae can lead to community-acquired gas gangrene in diabetic patient, which deserves clinical attention.