Objective To investigate the clinical distribution,antimicrobial resistance,carbapenemase resistance genes,virulence genes,capsular serotypes and ST subtypes of carbapenem-resistant Klebsiella pneumoniae(CRKP)strains in intensive care unit of a tertiary hospital in Hunan Province for better management of CRKP infections.Methods CRKP strains were isolated from 8 intensive care units of the First People's Hospital of Chenzhou City from January 2020 to December 2021.The isolates were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and tested by VITEK Compact 2 for antimicrobial susceptibility.Carbapenemase phenotype was detected by modified carbapenem inactivation method(mCIM).The capsular serotypes were determined by wzi sequencing.Carbapenem resistance genes and virulence genes were identified by PCR and Sanger sequencing.The strains were also analyzed by multilocus sequence typing(MLST)in terms of ST subtypes.Results The 75 CRKP strains were mainly isolated from geriatric ICU(28.0％)and neurosurgery ICU(20.0％).Overall,6.7％(5/75)and 16.0％(12/75)of the CRKP strains were resistant to tigecycline and ceftazidime-avibactam,respectively.The CRKP strains(＞96.0％resistant)were highly resistant to carbapenems,cephalosporins,β-lactam/β-lactamase inhibitor combinations,and levofloxacin.PCR and sequencing analysis found blaKPC-2 gene in 61 strains(81.3％),blaNDM-1 gene in 11 strains(14.7％),blaNDM-5 gene in 1 strain(1.3％),and blaOXA-48 gene in 2 strains(2.7％).MLST revealed that ST11(54.7％,41/75),ST1883(13.3％,10/75),and ST307(6.7％,5/75)were the top three ST subtypes.All ST11 and ST1883 CRKP strains harbored blaKPC-2.KL64(38.7％,29/75)and KL47(25.3％,19/75)were the most prevalent capsular serotypes among the 75 CRKP strains.The most common virulence genes among these CRKP strains were rmpA2(48.0％,36/75),iroN(38.7％,23/75)and iucA(37.3％,15/75).Conclusions The CRKP strains isolated from the intensive care units were mainly ST11-KL64 and ST11-KL47 types.Most of the strains harbor blaKPC-2 and virulence gene,and associated with high level antimicrobial resistance.It is urgent to strengthen the monitoring of molecular epidemiological characteristics of CRKP in order to inform individualized and precision treatment of CRKP infections.

Objective To investigate the influence of stress on insomnia among plateau soldiers,and explore the mediating role of parent-child alienation and the moderating role of emotion regulation strategies in this relationship.Methods A cross-sectional study design was adopted in this study.Our Self-designed General Information Questionnaire,Plateau Soldiers Stressor Scale,Insomnia Severity Index,Inventory of Alienation towards Parents,and Emotion Regulation Scale were employed to survey 2 065 plateau soldiers at an average altitude of 4 200 m.With aid of SPSS 26.0 statistics,descriptive analysis,correlation analysis and regression analysis were applied to analyze stress status,parent-child alienation,insomnia and emotion regulation in the participants,and then a structural equation model was established.Results ① The scores of stress,parent-child alienation and insomnia were significantly higher in the participants who were older,well educated(P＜0.01)and were or had been in a marital relationship(married,divorced,or widowed)(P＜0.001)than those without above conditions.② Significant pairwise positive correlations were observed among stress,parent-child alienation,insomnia,cognitive reappraisal,and expressive suppression(r=0.08～0.61,P＜0.001).③ Stress and expressive suppression could positively predict parent-child alienation(β=0.14～0.43,P＜0.001),and stress,parent-child alienation and expressive suppression could positively predict insomnia(β=0.04～0.25,P＜0.01).Cognitive reappraisal could not significantly predict parent-child alienation or insomnia.④ Parent-child alienation towards parents partially mediated the relationship between stress and insomnia,with a mediation effect accounting for 7.29％of the total effect.⑤ Expressive suppression moderated the impact of stress on insomnia(β=-0.01,P＜0.001),and the higher expressive suppression level mitigated the adverse effect of stress on insomnia when compared with the lower expressive suppression level.Conclusion For plateau soldiers,stress significantly predicts insomnia,with parent-child alienation playing a mediating role.Furthermore,a higher level of expressive suppression can effectively buffer the influence of stress on insomnia.

Objective:To investigate the effect of early external diaphragm pacing on the diaphragm function and prognosis of patients with mechanical ventilation.Methods:A total of 47 patients receiving invasive mechanical ventilation in the Emergency Intensive Care Unit of Hunan Provincial People's Hospital (The First Affiliated Hospital of Hunan Normal University) from October 2019 to July 2021 were selected and randomly divided into the treatment group ( n=23) and control group ( n=24). The patients in the control group received routine clinical treatment. On this basis, the treatment group received external diaphragm pacing treatment every day from the third day of mechanical ventilation until weaning, and was followed up to 30 days after discharge. The diaphragm thickness, diaphragm excursion, diaphragm thickening fraction, mechanical ventilation time, number of weaning failures, length of hospital stay and death toll were compared between the two groups. Results:Compared with the control group, the diaphragm thickness [(0.21±0.05) cm vs. (0.16±0.05) cm], diaphragm excursion [(1.38±0.37) cm vs. (1.11±0.48) cm], and diaphragm thickening fraction [26% (19%, 32%) vs. 18.5% (10.25%, 20%)] in the treatment group increased significantly (all P<0.05). The mechanical ventilation time was shorter in the treatment group [10 (7, 15) d vs. 13 (10.25, 19) d], and the difference was statistically significant ( P<0.05). There were no significant differences in the number of weaning failures (7 vs. 9), length of hospital stay [22 (15 , 30) d vs.. 24 (17.25, 34.25) d] and deaths (8 vs. 8) (all P>0.05). Conclusions:Early application of external diaphragm pacing can improve the diaphragm function of patients with mechanical ventilation, delay the decline in diaphragm function, increase diaphragm excursion and diaphragm thickening fraction, and shorten the mechanical ventilation time.

Objective : To investigate the effect of ginsenoside Rg1 (GRg1) on human retinal microvascular endothelial cells (HRMECs) glycolysis by regulating pyruvate kinase M2 ( PKM2) expression. Methods : HRMECs were cultured in vitro and divided into normal control (NC) group, high glucose (HG) group, high glucose + ginsenoside Rg1 (HG + GRg1) group, high glucose + ginsenoside Rg1 + low expression PKM2 ( HG + GRg1 + si-PKM2) group, and high glucose + ginsenoside Rg1 + overexpression PKM2 (HG + GRg1 + OE⁃PKM2) group. si-PKM2 and OE⁃PKM2 were transfected into HRMECs cells by cell transfection. The expression of PKM2 mRNA in HRMECs was detected by qRT⁃PCR. The expression levels of related proteins in HRMECs were detected by Western blot. The number of lumen formation in vitro was observed under an inverted microscope to quantify the angiogenesis ability. Cell culture medium of each group was collected, and glucose intake, lactate production and adenosine triphosphate(ATP)content were detected by glucose detection kit, lactate detection kit and ATP detection kit,re spectively. Results : HG induced HRMECs significantly increased the number of blood vessel formation, glycolysis and PKM2 expression, while GRg1 treatment significantly reduced the number of blood vessel formation, glycolysis and PKM2 expression; transfection of si⁃PKM2 assisted the inhibitory effect of GRg1 on glycolysis and angiogenesis while transfection of OE⁃PKM2 interfered with the function of GRg1 . Conclusion GRg1 inhibits angiogenesis by inhibiting PKM2 to reduce glycolysis of HRMECs.

Objective: To evaluate the efficacy and safety of 0.05% atropine eye drops for retarding myopia progression and ocular axial elongation in school children,and to provide a reference for the relevant prevention and control measures of myopia. Methods: A total of 188 children with myopia were randomly assigned to the experimental group(93) or to the control group(95). During the phase (first 24 months) I,children received treatment in each eye once a day. During the phase II (from 25th to the 36th month),no treatment was given. Standardized eye examinations including spherical equivalent(SE),axial length(AL),intraocular pressure(IOP) and potential atropine-related side effect assessment were performed every 6 months. Results: In phase I, the annual progression rates of equivalent spherical degree [(-0.35±0.21)D/year] and axial length [(0.11±0.07)mm/year] in the experimental group were significantly lower than those in the control group [(-0.83±0.26)D/year and (0.37±0.22)mm/year] (P<0.05). After withdrawal of atropine eye drops (phase II), the equivalent spherical degree progression rate [(-0.40±0.29)D/year] and axial length progression rate [(0.10±0.04)mm/year] in the experimental group were significantly lower than those in the control group [(0.73±0.40)D/year and (0.30±0.11)mm/year]. No serious adverse events associated with atropine were found during the follow up period. After the withdrawal of atropine, the pupil size, near visual acuity and adjustment gradually returned to the pre-treatment level. Conclusion 0.05% atropine eye drops may not only maintain the efficacy and reduce potential side effects of atropine but also significantly increase the compliance of children,0.05% atropine is a safe and effective treatment for retarding myopic progression in moderate myopia.

Objective:To explore the pathogenic spectrum and epidemiological characteristics of respiratory viruses in children with acute lower respiratory tract infection in Changsha, and provide scientific basis for disease prevention and treatment.Methods:A total of 1 092 respiratory tract specimens of children were collected and 12 respiratory viruses were detected by real-time quantitative transcription polymerase chain reaction.Results:Among the samples from 1 092 cases, those from 437 cases (40%) were positive for respiratory syncytial virus (RSV), 337 cases (30.9%) were positive for parainfluenza virus-3 (PIV-3), 263 cases (24.1%) were positive for human bocavirus (HBOV) and 228 cases (20.8%) were positive for adenovirus (ADV). The detection rates of boys and girls were 82.26% and 83.42%. The infection rate of RSV was higher in the group ≤ 6 months of age, the infection rates of PIV-3 and HBOV ≤2 years old were higher, and the infection rate of ADV was higher in the group between 6 months to 5 years old. The detection rates of virus infection of spring, summer, autumn and winter were 90.48%, 83.50%, 62.26% and 82.80%, respectively, there were significant differences among them.Conclusions:The main viruses in children with acute lower respiratory tract infection in Changsha were RSV and PIV-3. Mixed infections were common. Children under 2 years of age were more likely to get acute lower respiratory infections. Viruses had seasonal trends and peaked in winter and spring.

Objective: To investigate the characteristics and prognosis of clonal chromosomal abnormalities appearing in Philadelphia negative metaphases (CCA/Ph^-) cells in chronic myeloid leukemia (CML) with tyrosine kinase inhibitor (TKI) therapy. Methods: The clinical data of 30 cases with CCA/Ph^- during TKI treatment in Henan Cancer Hospital from August 2007 to July 2017 were retrospectively analyzed. The univariate factor was analyzed by Kaplan-Meier method. Multiple-factor was analyzed by Cox proportional risk model. Results: Of the 30 cases, 19 (63.3%) were males. At the first detection of CCA/Ph^- the median age was 44 (rang 14-68) years old and the median treatment of TKI was 13 (rang 2-94) months. The clones proportion of first detected CCA/Ph^-≥ 50% was found in 18 (60.0%) cases. TKI treatment for 3 months with BCR-ABL^IS less than 10% was seen in 14 (46.7%) patients. 63.3% (19/30) of CCA/Ph^- was transient (only one time) and 36.7% (11/30) was repeated (≥2 times) . Trisomy 8 dominant accounted for 60.0% (18/30) , -7/7q- for 13.3% (4/30) , loss of chromosome Y 6.7%. With a median of follow-up 50 months, 76.7% (23/30) cases were in complete cytogenetic response (CCyR) ; 63.3% (19/30) in major molecular response (MMR) , 43.3% (13/30) in undetectable minimal residual disease (UMRD) . The median event-free survival rate of (EFS) were 44 months, and 2-year and 5-year EFS were (82.1±7.3) % and (52.4±12.8) %, respectively. The median overall survival (OS) were 50 months, and 2-year and 5-year OS rates were (92.6±5.0) % and (77.2±14.7) %, respectively. Univariate analysis shows that the 2-year EFS of who in males, more than 2 times CCA/Ph^-, BCR-ABL^IS>10% at 3 months after TKI were significantly lower than women, transient CCA/Ph^-, and BCR-ABL^IS≤10% (P<0.05) . The 2-year OS rate in whom the occurrence frequency of CCA/Ph^- more than twice was significantly lower than those with transient CCA/Ph^- (P<0.05) . Multivariate analysis showed that CCA/Ph^- was an independent risk factor (RR=4.741, 95%CI 1.21-18.571, P=0.018) for EFS in CML patients. Conclusion Trisomy 8, -7/7q-, and -Y were the most common CCA/Ph^- during TKI treatment, with high clones proportion of ≥50%. CCA/Ph^- mainly occurred transiently or was permanent occasionally. CCA/Ph^- recurrence (≥2 times) was an independent risk factor for EFS and OS in CML with TKI.

Objective: To investigate the influence of additional clonal chromosome abnormalities in Ph negative cells (CCA/Ph^-) on the efficacy of chronic myeloid leukemia (CML) after tyrosine kinase inhibitors (TKI) treatment. Methods: The clinical data of 28 CML patients with CCA/Ph^- treated in Henan Cancer Hospital from July 2014 to December 2017 were analyzed retrospectively. The univariate analysis was carried out by Kaplan-Meier method. Multivariate analysis was done by Cox proportional risk model. Results: A total of 28 CCA/Ph^-patients were recruited including 17 males and 11 females with median age of 42.5 years old. The most common CCA/Ph^-were trisomy 8 (60.7%), monosomy 7 (14.3%). 64.3% CCA/Ph^-were transient and 35.7% recurrent (more than 2 times). Cytopenia in two or three lineages of peripheral blood was seen in 42.9% patients. As to the efficacy, 89.3% patients achieved major cytogenetic response (MCyR), 25% with major molecular response (MMR). The median follow-up time was 26.5 months. Treatment failure (TF) of TKI occurred in 32.1% patients with median duration of response 8 (1-41) months. Univariate analysis showed that TF rate was significantly correlated with the frequency of CCA/Ph^-and cytopenia (all P<0.05). The MMR rate was also significantly correlated with cytopenia (P<0.05). Cytopenia of two lineages or pancytopenia was an independent risk factor related to MMR rate (RR=3.868, 95%CI 1.216-12.298, P=0.022) . Conclusions Cytopenia in CCA/Ph^-appears to be an independent risk factor of MMR in CML patients with TKI treatment. The recurrent CCA/Ph^-may link to higher treatment failure rate. Drug withdrawal or alternative strategy should be considered according to response and the ABL kinase mutations.

Objective To investigate the clinical effect of pulmonary rehabilitation therapy including respiratory exercise and vibration expectoration on patients with pulmonary infection after abdominal surgery.Methods A retrospective case control study was conducted.Seventy-six patients with pulmonary infection after abdominal surgery admitted to the First Affiliated Hospital of Hunan Normal University from September 2015 to September 2016 were enrolled.According to whether accept the pulmonary rehabilitation therapy or not,the patients were divided into two groups.In the control group (n =35),the convemional expectoration method was adopted.The patients in pulmonary rehabilitation group (n =41) received both methods of the control group and pulmonary rehabilitation treatment,including respiratory exercise (effective cough,lip reduction breathing),respiratory exercise device (respiratory exerciser tri-ball),and vibrated expectoration.The 24-hour sputum volume,degree of comfort,inflammatory and pulmonary function parameters,and recovery situation were recorded in the two groups.Results ① There were no significant differences in the parameters of inflammation and pulmonary function before treatment between the two groups.After treatment,the white blood cell (WBC) and C-reactive protein (CRP) in both groups were significantly decreased,and the forced expiratory volume in 1 second (FEV1) and FEV1/forced vital capacity (FVC) were significantly increased.The above changes in pulmonary rehabilitation group were more significant than those of the control group [WBC (× 109/L):11.12 ± 2.88 vs.13.42 ± 2.62 at 3 days,8.22 ± 1.48 vs.9.27 ± 1.92 at 5 days;CRP (mg/L):13.47 ± 4.77vs.16.03±4.94 at 3 days,9.69±1.56 vs.11.77±1.41 at 5 days;FEV1 (L):2.48±0.14 vs.2.29±0.16 at 3 days,FEV1/FVC:0.78±0.04 vs.0.75±0.04 at 3 days;all P ＜ 0.05].② The 24-hour sputum volume within 3 days of pulmonary rehabilitation group were significantly higher than that of the control group (mL:30.51 ± 4.15 vs.18.30 ± 3.64at 1 day,31.08±3.22 vs.20.37±3.20 at 2 days,29.03±2.55 vs.19.03±2.51 at 3 days,all P ＜ 0.01].③ In the pulmonary rehabilitation group,the recovery time of pulmonary infection symptoms (days:5.44 ± 1.45 vs.6.20 ± 1.55),the days of antibiotic use (days:12.61 ± 3.15 vs.15.03 ± 3.78),the time of getting out of the bed (days:4.05 ± 0.74vs.4.51±0.89),and the hospital days (days:19.95±3.90 vs.22.00±4.42) were significantly shorter than those of the control group (all P ＜ 0.05),and the degree of comfort was significantly better than that of the control group (comfort score:2.71 ±0.90 vs.2.14±0.91,P ＜ 0.01).Conclusion The application of pulmonary rehabilitation including respiratory exercise and vibration expectoration in abdominal surgery patients with pulmonary infection can promote recovery,and it has a good clinical and practical application value.

Objective To study the curative efficacy and safety of single-unit umbilical cord blood transplantation (sUCBT) for malignant hematologic diseases,which is provided by China's public cord blood bank.Methods We retrospectively analyzed 409 cases of malignant hematologic diseases who accepted myeloablative single-unit unrelated donor UCBT without ATG at our center between May 2008 and December 2016.A comparative analysis was made on the total nuclear cells (TNC) of the umbilical cord blood before freezing and after thawing,the cells of CD34+,the recovery rate of cells and the clinical effect of UCBT.Result 409 units of umbilical cord blood used in UCBT respectively came from eight China's public cord blood banks.The average TNC of 409 units of umbilical cord blood before freezing and after the tubular recovery were respectively 18.5 × 108 and 16.34 × 108 (p =0.000).The average recovery rate of the tubular recovery was 88.5％,and there was significant difference among cord blood banks (P =0.000).The average TNC of umbilical cord blood before freezing and transfusion were respectively 18.5 × 108 and 15.86 × 108 (p =0.000).The average recovery rate of umbilical cord blood transfusion was 85.9％,with the difference being significant among cord blood banks (P =0.000).The average number of CD34+ cells before freezing and after the tubular recovery was 11.18 × 106and 8.68 × 106 (p =0.000).The average recovery rate of CD34+ cells after the tubular recovery was 80.75 ％,with the difference being significant among the cord blood banks (P =0.000).At 42nd day after UCBT,the cumulative incidence of neutrophil engraftment was 95.4％,and the median time of the engraftment was 17 days (11-38 days).The cumulative incidence of platelet engraftment at 120th day was 84.6％,and the median time of the engraftment was 36 days (14-93 days).The cumulative incidence of erythrocyte engraftment at 60th day was 92％,and the median time of engraftment was 22 days (9d-60 days).After the umbilical cord blood provided by each bank was used in UCBT,it got the difference in cumulative incidence of engraftment.The P values for cumulative incidence of neutrophil,platelet and erythrocyte engraftment were respectively 0.004,0.01 and 0.000 2,with the differences being statistically significant.At 100th day after UCBT,the cumulative incidence of Ⅱ-Ⅳ and Ⅲ-Ⅳ degrees of acute graft-versus-host disease (aGVHD) was respectively 28.63％ and 15.7％.After umbilical cord blood provided by each bank was used in UCBT,it got the difference in cumulative incidence of aGVHD.There was no significant difference between Ⅱ-Ⅳ and Ⅲ-Ⅳ degrees (P =0.809 and 0.68 respectively).At 3rd year after UCBT,the cumulative incidence of relapse was 15.89％.After umbilical cord blood provided by each bank was used in UCBT,there was no significant difference in the cumulative incidence of relapse (P =0.898).At 3rd year after UCBT,the overall survival (OS) rate and disease free survival (DFS) rate were respectively 66.7％ and 59％.After umbilical cord blood provided by each bank was used in UCBT,it got the difference in OS and DFS.There was no significant difference in OS and DFS (P =0.566 and 0.703 respectively).At 3rd year after sUCBT,the rate of graft-versus-host diseases/relapse-free survival (GRFS) was 54.3％.After umbilical cord blood provided by each bank was used in UCBT,there was no significant difference in the rate of GRFS (P =0.449).Conclusion The umbilical cord blood provided by China's public cord blood bank was used in UCBT.It has a high safety and good efficacy in treating malignant hematologic diseases.But it needs to set up the standardized and normalized quality-control system of umbilical cord blood for China's public cord blood bank.