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 risk factors for yersiniosis, so as to provide insights into prevention of yersiniosis. Methods: The patients with yersiniosis admitted to the clinics in the surveillance site of Chengbei Township of Jin'an District and Chengnan Township of Yu'an District in Lu'an City from 2013 to 2021 were included as the case group, and the healthy family members matched to cases were selected as the family control group, while normal residents with a 1︰2 match in the same village, gender, and age difference within 5 years were included in the community control group. Participants' demographics, hand-washing and eating habits, living environment hygiene, poultry and livestock feeding were collected using questionnaire surveys, and factors affecting yersiniosis were identified using a multivariable conditional logistic regression model. Results: There were 43 cases in the case group, with a median (interquartile range) age of 45 (34) years, 91 cases in the family control group, with a median (interquartile range) age of 36 (36) years and 86 cases in the community control group, with a median (interquartile range) age of 46 (34) years. Multivariable conditional logistic regression analysis showed that compared with the family control group, the habit of drinking unboiled water (OR=6.721, 95%CI: 1.765-25.588), and direct consumption of food stored in the refrigerator (OR=7.089, 95%CI: 1.873-26.829) were risk factors for yersiniosis in the case group; and compared with the community control group, not washing hands after contacting with poultry and livestock (OR=50.592, 95%CI: 2.758-927.997), habit of eating raw vegetables and fruits (OR=5.340, 95%CI: 1.022-27.887), direct consumption of food stored in the refrigerator (OR=19.973, 95%CI: 2.118-188.336), and unclean refrigerator (OR=12.692, 95%CI: 1.992-80.869) were risk factors for yersiniosis in the case group. Compared with the family and community control groups, not washing hands after contacting with poultry and livestock (OR=4.075, 95%CI: 1.427-11.637), habit of drinking unboiled water (OR=4.153, 95%CI: 1.331-12.957), habit of eating raw vegetables and fruits (OR=4.744, 95%CI: 1.609-13.993), and direct consumption of food stored in the refrigerator (OR=5.051, 95%CI: 1.773-14.395) were risk factors for yersiniosis in the control group. Conclusion Unhealthy habits such as eating raw vegetables and fruits, drinking unboiled water, direct consumption of food stored in the refrigerator, unclean refrigerator, and not washing hands after contacting poultry and livestock may increase the risk of yersiniosis.

Objective: To construct a model for clinical identification of spotted fever (SF) and severe fever with thrombocytopenia syndrome (SFTS), so as to provide insights into early identification of SF and SFTS. Methods: The clinical data of laboratory-confirmed SF and SFTS patients in secondary and tertiary hospitals in Lu'an City, Anhui Province from May 2017 to May 2021 were retrieved from Chinese Disease Prevention and Control Information System. Factors affecting SF were identified using a logistic regression model, and the model for early identification of SF and SFTS was created. The model fitting effect was evaluated using Hosmer-Lemeshow test, and the value of the model for identification of SF and SFTS was evaluated using the area under the receiver operating characteristic curve (AUC). Results: Data of 62 SF cases and 115 SFTS cases were included. Multivariable logistic regression analysis showed that rash (β=5.994), C-reactive protein (β=4.409), white blood cell (β=-3.176) and platelet (β=-3.234) were included in the model, which were scored 6, 4, -3 and -3, with a total score ranging from -5 to 10. Hosmer-Lemeshow test revealed a high model fitting effect (χ2=3.245, P=0.662). The AUC of the model was 0.992, and the sensitivity and specificity were 0.935 and 0.991 if the cutoff was 1. Conclusion A model for early identification of SF and SFTS that includes four variables of rash, C-reactive protein, white blood cell and platelet has been created, which has a high accuracy.

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 clinical characteristics and prognostic values of TP53 gene variant in patients with acute leukemia(AL). METHODS: The clinical data of 44 newly diagnosed AL patients with TP53 variant detected by next generation sequencing (NGS) were analyzed retrospectively. Targeted sequencing technique containing 108 leukemia-related genes was used for variant analysis, and conventional R-banding technique was used for karyotype analysis. The clinical features, cytogenetics, gene variant, curative effect and survival of AL patients with TP53 gene variant were analyzed. RESULTS: The median age of AML patients with TP53 gene variant (46 years) was higher than that of ALL patients (17.5 years), and the median number of bone marrow blasts (40.5%) was lower than the latter (89.2%), the differences were statistically significant (P< 0.01). A total of 28 cases of abnormal karyotype were detected, of which 25 cases were complex karyotype, 16 cases were monomeric karyotype, 14 cases had -17/17p-. The detection rates of TP53 in complex karyotype, monomeric karyotype and -17/17p- were 59.5%, 38.1% and 33.3%, respectively. Subgroup analysis showed that the detection rate of TP53 gene abnormalities in AML and ALL complex karyotypes was 73.1% and 40% respectively, the difference was statistically significant. A total of 41 TP53 gene variant types were found, and the median variant frequency was 43.58%. 75.6% variant was located in the DNA binding domain. The concomitant variant genes were mainly TET2 and IKZF1. Among 18 AML and 17 ALL patients who could be evaluated the curative effect, the CR rate of one course of treatment was 22.2% and 94.12% respectively, and the difference was statistically significant. The median RFS of 4 cases of AML with CR and 16 cases of ALL with CR were 174 and 246 days respectively, the difference was statistically insignificant. The median OS of AML and ALL was 20 and 375 days respectively, the difference was statistically significant. CONCLUSION The TP53 gene variant is associated with the complex karyotype of AML, but has no significant effect on ALL. The variant site of TP53 gene was mainly distributed in the DNA binding domain. The remission rate of AML with TP53 gene variant was lower than that of ALL. The prognosis of AL patients with TP53 gene variant is poor, so allogeneic hematopoietic stem cell transplantation should be performed as soon as possible to prolong the survival of the patients.
Acute Disease ; Humans ; Leukemia, Myeloid, Acute/genetics* ; Middle Aged ; Mutation ; Retrospective Studies ; Tumor Suppressor Protein p53/genetics*

Objective:To explore the characteristics and clinical significance of clonal heterogeneity in patients with acute lymphoblastic leukemia(ALL).Methods:From January 2016 to June 2019, 170 newly diagnosed ALL patients were enrolled in the Department of Hematology, Henan Cancer Hospital, including 93 males and 77 females, with a median age of 17 (2-80) years. Fifty-two ALL-related genes were detected by high-throughput sequencing technique. The clonal heterogeneity of mutations was analyzed according to the variant allele frequency (VAF) and the results of flow cytometry. The prognostic value of mutations was also evaluated.Results:Gene mutations were detected in 121 (71.2%, 121/170) patients, of which 2 or more clones were detected in 18 (52.9%, 18/34) T-cell acute lymphoblastic leukemia patients, while only 23 (16.9%, 23/136) B-cell acute lymphoblastic leukemia patients were positive of multiple mutations ( P<0.01).Gene mutation-related clonal heterogeneity analysis showed that 2 or more clones were frequent in patients with NOTCH1 mutations (13/19 patients) ( P<0.01). Event free survival (EFS) in patients with 3 or more clones was significantly lower than other patients (χ 2=10.330, P=0.016). Child ALL patients had similar result, that multiple clones predicted lower overall survival (OS) and EFS (OS: χ 2=7.974, P=0.047; EFS: χ 2=10.860, P=0.013). Conclusion:Clonal heterogeneity in ALL patients is closely related to the different origin of lymphocyte lineages and the age of onset, which may reveal the nature of the disease and predict the clinical outcome.

Objective: To explore the relationship between driver gene mutation (JAK2, MPL and CALR) and disease type in BCR-ABL negative myeloproliferative neoplasms (MPNs) including primary myeloid fibrosis (PMF), essential thrombocytosis (ET) and polycythemia vera (PV). Methods: A total of 32 MPN related genes were detected by high-throughput sequencing in 156 MPN patients. The relationships between disease type and patients′ general performance, the characteristics of driver gene mutations, concomitant gene mutations were analyzed. Results: In the population with JAK2 V617F positive mutation, the proportion of patients over 60 years old in PMF was higher than that with ET or PV. By high-throughput sequencing, 22 concomitant gene mutations were detected in 46 patients with JAK2, MPL or CALR mutations, including 4 (8.3%) in PV, 20 (29.4%) in ET, and 22 (55.0%) in PMF. DNMT3A mutation was detected only in patients with PV, while splicing factor related genes including SF3B1, SRSF2 and U2AF1 were only accompanied by PMF. According to the variation allele frequency (VAF) value of JAK2 V617F mutation, the VAF value associated with PV was the highest (68.15%), followed by PMF (37.7%) and ET (23%). However, there were significant differences in the incidence of JAK2 V617F homozygous among 3 different diseases. In patients with JAK2 mutation, the proportion of other gene mutations in PV and ET was significantly lower than that in PMF. Conclusions Under the condition of common driver gene mutations (JAK2, MPL and CALR), patients′ age, VAF value and homozygous state, concomitant gene mutations are closely related to different disease type. These correlations help to improve clinical understanding of disease characteristics and risk assessment.

OBJECTIVE: To explore the mechanism of METHODS: Healthy male DBA/1 mice were used for CIA modeling. Twenty-five CIA mice with successful modeling and similar arthritis index (AI) scores were randomized equally into model group (CIA), methotrexate (MTX) group, and low-, medium-, and high-dose XWGD groups (0.975, 1.95, and 3.9 g/mL, respectively), with another 5 normal mice as the normal control group. The mice in normal control and CIA groups were given saline once a day, those in MTX group were given 0.1 mg/mL MTX once a week, and those in XWGD groups were treated daily via garage of XWGD containing crude drugs of different doses for 28 consecutive days. The AI score and HE staining were used to evaluate the changes in the joints of the CIA mice. The effect of XWGD on Th1, Th17, MDSC, G-MDSC and M-MDSC cells were evaluated with flow cytometry. RESULTS: Treatment with MTX and different doses of XWGD significantly decreased the AI score of the mice and relieved joint inflammation as compared with the model group ( CONCLUSIONS XWGD can improve joint inflammation in CIA mice by increasing the percentages of G-MDSC cells and decreasing the percentages of M-MDSC, Th1 and Th17 cells, and a high dose of XWGD can produce an equivalent therapeutic effect to methotrexate but with better safety.
Animals ; Arthritis, Experimental/drug therapy* ; Arthritis, Rheumatoid/drug therapy* ; Male ; Methotrexate ; Mice ; Mice, Inbred DBA ; Th17 Cells

Objective:To understand sexual needs and factors of risky sexual behaviors among elderly men at different ages in two communities of Qiandongnan Miao and Dong autonomous prefecture and provide basis for targeted HIV prevention and intervention.Methods:Two communities in the prefecture were selected as study sites. Questionnaire surveys were carried out among elderly men aged 50 and over who visited or consulted in the communities from June to December 2018, and they were tested for HIV and syphilis antibodies.Results:Among 400 elderly men, 209 (52.2%) were 50-64 years old, and 191(47.8%) were above 65 years old. They were mainly Miao people, accounting for 66.3% (265/400), and 235 (58.8%) had an education no more than 6 years. HIV awareness of the two age groups were only 25.8% (54/199) and 26.2% (50/191), respectively. Among those aged 50-64, 142 (68.0%) felt normal sexual desire, and 153 (73.6%) reported penile erections or erections in most cases whenever sex, and 52.9% (110) ejaculated most of the time. HIV prevalence was 1.0% (4/400). Compared with the over 65-year-old group, the proportion of having sex with spouse/stable partners (89.5%, 179/200), proportion of no condom use with their spouse/stable sexual partners during the most recent sex (93.8%, 168/179), proportion of having casual sex (11.0%, 23/209) and commercial sex (3.8%, 8/209) were all higher among 50-64 age group. In comparison to those aged over 65 years old, average monthly income>3 000, and use of sex helper, aged 50-64 ( OR=2.70, 95% CI: 1.22-5.95), average monthly income ≤1 000 yuan ( OR=2.79, 95% CI: 1.25-6.21), and no use of sex helper ( OR=3.78) (95% CI: 1.65-8.67) were related factors of HIV risky sexual behavior last time. Conclusion:Elderly men in the minority prefecture had low HIV awareness. Compared with those≥65 years old, the 50-64 age group had more active sexual behaviors and higher sexual needs. Those from 50-64 age group, with lower economic level and good sexual ability were more likely to have HIV risky sexual behaviors.