Objective To observe the change of the number of endothelial progenitor cells(EPCs)in peripheral circulation after acute middle cerebral artery occlusion/reperfusion(MCAO/R)and to evaluate the therapeutic effect of VEGF through mobilizing bone marrow-derived EPCs in treatment of mouse brain infarction after acute MCAO/R.MethodsTotally 36 mice were randomized into MCAO/R+VEGF group,MCAO/R group and sham operation group.MCAO/R mice model was established according Longa's method.VEGF [3.3 ng/(g?d),for 7 d] was injected intraperitoneally to the mice of MCAO/R+VEGF group to mobilize bone marrow-derived EPCs.The other 2 group received an injection of normal saline.At days 1,4,7 during mobilization,neurological functions were evaluated and blood samples were taken from angular vein.Then the number of EPCs in peripheral circulation in MCAO/R group and MCAO/R+VEGF group was detected by flow cytometry.Mice were decapitated and brains sliced and stained with triphenyltetrazolium chloride(TTC)to calculate infarct volume using specific image analyzing system.Infarct volumes were calculated and compared among groups.ResultsThe number of EPCs in MCAO/R+VEGF group began to increase at day 1 after treatment,and peaked at day 4 and sustained to day 7,which was significantly larger than those in MCAO/R group and sham operation group at every time point(P

Objective: To investigate the prevalence of dyslipidemia among adults in Moyu County, Hotan Prefecture, Xinjiang Uygur Autonomous Region, so as to provide insights into the management of dyslipidemia. Methods: The physical examination data of permanent residents at ages of 18 years and older were collected from Moyu County from 2018 to 2019, including demographic features, height, body weight, blood pressure and blood biochemical parameter measurements. The epidemiological characteristics of dyslipidemia were analyzed among residents. Results: Totally 166 142 adults were investigated, with a mean age of (41.08±15.72) years. There were 77 744 men (46.76%), 34 728 obese adults (20.90%), 5 776 adults with diabetes (3.48%) and 26 294 adults with hypertension (15.83%). The levels of triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) were (1.38±0.98), (4.13±1.19), (1.31±0.41) and (2.25±0.77) mmol/L, respectively. The prevalence of dyslipidemia was 29.72%, and the detection rates of high TG, high TC, low-HDL-C and high LDL-C were 9.73%, 4.59%, 19.65% and 1.99%, respectively. The prevalence of dyslipidemia was higher in men than in women (33.97% vs. 25.98%; χ2=1 264.729, P<0.001), and the prevalence increased with age (χ2=539.382, P<0.001) and body mass index (χ2=3 601.833, P<0.001). The prevalence of dyslipidemia was 35.48% among patients with hypertension (χ2=497.251, P<0.001) and 46.07% among patients with diabetes (χ2=766.142, P<0.001). There were 9 059 adults with two and more abnormal blood lipid indexes (18.35%), and the detection rates of two and more abnormal blood lipid indexes were 26.50%, 24.14% and 33.15% among patients with obesity, hypertension and diabetes, respectively. Conclusions The prevalence of dyslipidemia among adults in Moyu County is lower than the national level, and low-HDL cholesterolemia is the most common type of dyslipidemia. The prevalence of dyslipidemia is high among patients with obesity, hypertension and diabetes mellitus, in whom abnormality of multiple blood lipid indexes is measured.

Objective To summarize the outcomes and prognostic factors in esophageal cancer (EC) patients.Methods A total of 307 EC patients of stages Ⅱ-Ⅲ were treated with concurrent chemoradiotherapy in our hospital from September 2006 to July 2014.There were 73 patients with stage Ⅱ and 234 with stage Ⅲ.The radiotherapy dose was 50-70 Gy (median 60 Gy).Concurrent chemoradiotherapy were used with fluorouracil plus platinum (PF,166),paclitaxel plus platinum (TP,82) or platinum only (P,59).The Kaplan-Meier method was used to calculate overall survival (OS) and progression-free survival (PFS) rates,the log-rank test was used for survival difference analysis and univariate prognostic analysis.The Cox regression model was used for multivariate prognostic analysis.Results The 1-,3-5-year OS and PFS rates were85.6％,53.8％,36.9％ and 74.6％,43.7％,33.1％,respectively.The median OS and PFS were 41.6 months and 29.8 months.The univariate analysis indicated that T stage,N stage,clinical stage,lesion location,lesion length and chemotherapy regimen were prognostic factors for OS and PFS (P=0.007 and 0.013,0.000 and 0.000,0.000 and 0.000,0.002 and 0.000,0.141 and 0.005,0.018 and 0.165).Multivariate analysis showed that T stage,N stage,lesion location and chemotherapy regimen were prognostic factors for OS (P =0.024,0.000,0.007 and 0.028),lesion location,lesion length and N stage were prognostic factors for PFS (P=0.004,0.033 and 0.035).The median OS and PFS for EC patients treated by total dose 50-60 Gy,＞60-70 Gy were 47.4 months,37.8 months (P=0.469) and34.1 months,25.1 months (P=0.0.233),therewere no statistic difference.Conclusions The outcome of EC patients treated with concurrent chemoratherapy could obtain a long-term survival,combination chemotherapy is superior to single drug,there are no statistical difference between high-dose and low-dose,and the acute toxic effects can be tolerated.

Background: Brucella infection induces brucellosis, a zoonotic disease. The intracellular circulation process and virulence of Brucella mainly depend on its type IV secretion system (T4SS) expressing secretory effectors. Secreted protein BspJ is a nucleomodulin of Brucella that invades the host cell nucleus. BspJ mediates host energy synthesis and apoptosis through interaction with proteins. However, the mechanism of BspJ as it affects the intracellular survival of Brucella remains to be clarified. Objectives: To verify the functions of nucleomodulin BspJ in Brucella's intracellular infection cycles. Methods: Constructed Brucella abortus BspJ gene deletion strain (B. abortus ΔBspJ) and complement strain (B. abortus pBspJ) and studied their roles in the proliferation of Brucella both in vivo and in vitro. Results: BspJ gene deletion reduced the survival and intracellular proliferation of Brucellaat the replicating Brucella-containing vacuoles (rBCV) stage. Compared with the parent strain, the colonization ability of the bacteria in mice was significantly reduced, causing less inflammatory infiltration and pathological damage. We also found that the knockout of BspJ altered the secretion of cytokines (interleukin [IL]-6, IL-1β, IL-10, tumor necrosis factor-α, interferon-γ) in host cells and in mice to affect the intracellular survival of Brucella. Conclusions BspJ is extremely important for the circulatory proliferation of Brucella in the host, and it may be involved in a previously unknown mechanism of Brucella's intracellular survival.

OBJECTIVE: To detect the mutation site in a pedigree affected with autosomal dominant polycystic kidney disease (ADPKD) and verify its impact on the protein function. METHODS: Peripheral blood samples were collected from the proband and his pedigree members for the extraction of genomic DNA. Mutational analysis was performed on the proband through whole-exome sequencing. Suspected variant was verified by Sanger sequencing. A series of molecular methods including PCR amplification, restriction enzyme digestion, ligation and transformation were also used to construct wild-type and mutant eukaryotic expression vectors of the PKD2 gene, which were transfected into HEK293T and HeLa cells for the observation of protein expression and cell localization. RESULTS: The proband was found to harbor a c.2051dupA (p. Tyr684Ter) frame shift mutation of the PKD2 gene, which caused repeat of the 2051st nucleotide of its cDNA sequence and a truncated protein. Immunofluorescence experiment showed that the localization of the mutant protein within the cell was altered compared with the wild-type, which may be due to deletion of the C-terminus of the PKD2 gene. CONCLUSION The c.2051dupA (p. Tyr684Ter) mutation of the PKD2 gene probably underlay the pathogenesis of ADPKD in this pedigree.
DNA Mutational Analysis ; Female ; Frameshift Mutation ; HEK293 Cells ; HeLa Cells ; Humans ; Male ; Pedigree ; Polycystic Kidney, Autosomal Dominant/physiopathology* ; Protein Kinases/genetics* ; Protein Transport/genetics* ; Whole Exome Sequencing

OBJECTIVE: To analyze the pathogenic variant of preaxial polydactyly in a Chinese Han pedigree and identify the cause of polydactyly. METHODS: The peripheral blood DNA of the proband and her parents was extracted. The polydactyly-related genes were detected by trio whole exome sequencing, and the suspected pathogenic gene was screened out. Sanger sequencing was applied to other members of the pedigree. RESULTS: The results of gene sequencing showed that the LMBR1 gene had a heterozygous variant of c.423+4909(IVS5)C>T in 6 patients of the pedigree. The same variant was not detected in family members with normal phenotype. Based on the ACMG guidelines, c.423+4909(IVS5)C>T of the LMBR1 gene was predicted to be pathogenic (PM1+PM2+PP1-S(PS)+PP4+PP5). CONCLUSION The heterozygous C>T variant at position 4909 of intron 5 of the LMBR1 gene probably underlies the disease in this pedigree.
China ; Female ; Humans ; Mutation ; Pedigree ; Polydactyly/genetics* ; Thumb ; Whole Exome Sequencing

OBJECTIVE: To explore the genetic basis for a Chinese patient with amyotrophic lateral sclerosis (ALS). METHODS: Peripheral blood samples were collected from the patient and his parents for the extraction of genomic DNA. Genetic variant was identified by whole exome sequencing. Candidate variant was verified by Sanger sequencing of his parents and healthy controls. RESULTS: The patient was found to harbor a heterozygous c.420C>G (p.Asn140Lys) variant of the SOD1 gene. The same variant was not detected in his parents and 100 healthy controls. The variant has not been included in HGMD, dbSNP and other databases. CONCLUSION The c.420C>G variant of the SOD1 gene may underlie the ALS in this patient. Above finding has enriched the spectrum of SOD1 gene variants.
Amyotrophic Lateral Sclerosis/genetics* ; China ; Heterozygote ; Humans ; Superoxide Dismutase-1/genetics* ; Whole Exome Sequencing

BACKGROUND: Epidermal growth factor receptor (EGFR) mutations are often associated with non-EGFR genetic alterations, which may be a reason for the poor efficacy of EGFR tyrosine kinase inhibitors (TKIs). Here we conducted this study to explore whether EGFR-TKIs combined with chemotherapy would benefit advanced lung adenocarcinoma patients with both sensitive EGFR mutation and concomitant non-EGFR genetic alterations. METHODS: Cases of advanced lung adenocarcinoma with EGFR mutation combined with concomitant non-EGFR genetic alterations were retrospectively collected. And the patients were required to receive first-line EGFR-TKIs and chemotherapy combination or EGFR-TKIs monotherapy. Demographic, clinical and pathological data were collected, and the electronic imaging data were retrieved to evaluate the efficacy and time of disease progression. Survival data were obtained through face-to-face or telephone follow-up. The differences between the two groups in objective response rate (ORR), disease control rate (DCR), progression-free survival (PFS) and overall survival (OS) were investigated. RESULTS 107 patients were included, including 63 cases in the combination group and 44 cases in the monotherapy group. The ORR were 78% and 50% (P=0.003), and DCR were 97% and 77% (P=0.002), respectively. At a median follow-up of 13.7 mon, a PFS event occurred in 38.1% and 81.8% of patients in the two groups, with median PFS of 18.8 mon and 5.3 mon, respectively (P<0.000,1). Median OS was unreached in the combination group, and 27.8 mon in the monotherapy group (P=0.31). According to the Cox multivariate regression analysis, combination therapy was an independent prognostic factor of PFS CONCLUSIONS: In patients with EGFR-mutant advanced lung adenocarcinoma with concomitant non-EGFR genetic alterations, combination of TKIs and chemotherapy was significantly superior to EGFR-TKIs monotherapy, which should be the preferred treatment option.
Adenocarcinoma of Lung/genetics* ; ErbB Receptors/genetics* ; Humans ; Lung Neoplasms/pathology* ; Mutation ; Protein Kinase Inhibitors/therapeutic use* ; Retrospective Studies

Objective: To explore the clinical characteristics and prognosis of the new coronavirus 2019-nCoV patients combined with cardiovascular disease (CVD). Methods: A retrospective analysis was performed on 112 COVID-19 patients with CVD admitted to the western district of Union Hospital in Wuhan, from January 20, 2020 to February 15, 2020. They were divided into critical group (ICU, n=16) and general group (n=96) according to the severity of the disease and patients were followed up to the clinical endpoint. The observation indicators included total blood count, C-reactive protein (CRP), arterial blood gas analysis, myocardial injury markers, coagulation function, liver and kidney function, electrolyte, procalcitonin (PCT), B-type natriuretic peptide (BNP), blood lipid, pulmonary CT and pathogen detection. Results: Compared with the general group, the lymphocyte count (0.74×10⁹ (0.34×10⁹, 0.94×10⁹)/L vs. 0.99×10⁹ (0.71×10⁹, 1.29×10⁹)/L, P=0.03) was extremely lower in the critical group, CRP (106.98 (81.57, 135.76) mg/L vs. 34.34 (9.55,76.54) mg/L, P<0.001) and PCT (0.20 (0.15,0.48) μg/L vs. 0.11 (0.06,0.20)μg/L, P<0.001) were significantly higher in the critical group. The BMI of the critical group was significantly higher than that of the general group (25.5 (23.0, 27.5) kg/m² vs. 22.0 (20.0, 24.0) kg/m², P=0.003). Patients were further divided into non-survivor group (17, 15.18%) group and survivor group (95, 84.82%). Among the non-survivors, there were 88.24% (15/17) patients with BMI> 25 kg/m², which was significantly higher than that of survivors (18.95% (18/95), P<0.001). Compared with the survived patients, oxygenation index (130 (102, 415) vs. 434 (410, 444), P<0.001) was significantly lower and lactic acid (1.70 (1.30, 3.00) mmol/L vs. 1.20 (1.10, 1.60) mmol/L, P<0.001) was significantly higher in the non-survivors. There was no significant difference in the proportion of ACEI/ARB medication between the critical group and the general group or between non-survivors and survivors (all P>0.05). Conclusion COVID-19 patients combined with CVD are associated with a higher risk of mortality. Critical patients are characterized with lower lymphocyte counts. Higher BMI are more often seen in critical patients and non-survivor. ACEI/ARB use does not affect the morbidity and mortality of COVID-19 combined with CVD. Aggravating causes of death include fulminant inflammation, lactic acid accumulation and thrombotic events.

Flagella are the main motility structure of Clostridioides difficile that affects the adhesion, colonization, and virulence of C. difficile in the human gastrointestinal tract. The FliL protein is a single transmembrane protein bound to the flagellar matrix. This study aimed to investigate the effect of the FliL encoding gene flagellar basal body-associated FliL family protein (fliL) on the phenotype of C. difficile. The fliL gene deletion mutant (ΔfliL) and its corresponding complementary strains (: : fliL) were constructed using allele-coupled exchange (ACE) and the standard molecular clone method. The differences in physiological properties such as growth profile, antibiotic sensitivity, pH resistance, motility, and spore production ability between the mutant and wild-type strains (CD630) were investigated. The ΔfliL mutant and the : : fliL complementary strain were successfully constructed. After comparing the phenotypes of strains CD630, ΔfliL, and : : fliL, the results showed that the growth rate and maximum biomass of ΔfliL mutant decreased than that of CD630. The ΔfliL mutant showed increased sensitivity to amoxicillin, ampicillin, and norfloxacin. Its sensitivity to kanamycin and tetracycline antibiotics decreased, and the antibiotic sensitivity partially returned to the level of CD630 strain in the : : fliL strain. Moreover, the motility was significantly reduced in the ΔfliL mutant. Interestingly, the motility of the : : fliL strain significantly increased even when compared to that of the CD630 strain. Furthermore, the pH tolerance of the ΔfliL mutant significantly increased or decreased at pH 5 or 9, respectively. Finally, the sporulation ability of ΔfliL mutant reduced considerably compared to the CD630 strain and recovered in the : : fliL strain. We conclude that the deletion of the fliL gene significantly reduced the swimming motility of C. difficile, suggesting that the fliL gene is essential for the motility of C. difficile. The fliL gene deletion significantly reduced spore production, cell growth rate, tolerance to different antibiotics, acidity, and alkalinity environments of C. difficile. These physiological characteristics are closely related to the survival advantage in the host intestine, which is correlated with its pathogenicity. Thus, we suggested that the function of the fliL gene is closely related to its motility, colonization, environmental tolerance, and spore production ability, which consequently affects the pathogenicity of C. difficile.
Humans ; Clostridioides/metabolism* ; Clostridioides difficile/metabolism* ; Bacterial Proteins/metabolism* ; Virulence ; Anti-Bacterial Agents/metabolism*