Objective:To explore the risk factors of severe acute kidney injury (AKI) in septic patients, and to establish an hour-specific prediction model.Methods:Based on the information of septic patients in the Medical Information Mart for Intensive Care-Ⅳ (MIMIC-Ⅳ) database, general information, comorbidities, vital signs, severity scoring system, laboratory indicators, invasive operations and medication use were recorded. The enrolled patients were randomized into a training set and a validation set according to a ratio of 7∶3. AKI was diagnosed according to the guidelines of Kidney Disease: Improving Global Outcome (KDIGO). Based on Lasso regression and Cox regression, the risk factors of severe AKI (AKI stage 2 and stage 3) in septic patients were analyzed and hour-specific prediction model were established. Consistency index (C-index), area under the receiver operator characteristic curve (AUC) and calibration curve were used to assess the predictive efficacy of the model.Results:A total of 20 551 septic patients were enrolled, including 14 385 patients in the training set and 6 166 patients in the validation set. Multivariate Cox regression analysis showed that atrial fibrillation [hazard ratio ( HR) = 1.266, 95% confidence interval (95% CI) was 1.150-1.393], heart failure ( HR = 1.348, 95% CI was 1.217-1.493), respiratory failure ( HR = 1.565, 95% CI was 1.428-1.715), heart rate ( HR = 1.004, 95% CI was 1.002-1.007), mean arterial pressure ( HR = 1.245, 95% CI was 1.126-1.377), lactic acid ( HR = 1.051, 95% CI was 1.025-1.077), simplified acute physiology score Ⅱ (SAPSⅡ, HR = 1.019, 95% CI was 1.016-1.023), serum creatinine ( HR = 1.171, 95% CI was 1.127-1.216), anion gap ( HR = 1.024, 95% CI was 1.010-1.038), serum potassium ( HR = 1.155, 95% CI was 1.079-1.236), white blood cell count ( HR = 1.006, 95% CI was 1.003-1.009) and furosemide use ( HR = 0.414, 95% CI was 0.368-0.467) were independently associated with severe AKI in septic patients (all P < 0.01). The above predictors were applied to construct an hour-specific prediction model for the occurrence of severe AKI in septic patients. The C-index of the prediction model was 0.723 and 0.735 in the training and validation sets, respectively. The AUC for the occurrence of severe AKI at 12, 24, and 48 hours were 0.795 (95% CI was 0.782-0.808), 0.792 (95% CI was 0.780-0.805), and 0.775 (95% CI was 0.762-0.788) in the training set, and the AUC were 0.803 (95% CI was 0.784-0.823), 0.791 (95% CI was 0.772-0.810), and 0.773 (95% CI was 0.752-0.793) in the validation set, respectively. The calibration curves of the two cohorts were in good agreement. Conclusion:The hour-specific prediction model effectively identifies high-risk septic patients for developing severe AKI within 48 hours, aiding clinicians in stratifying patients for early therapeutic interventions to improve outcomes.

Humans ; Blood Pressure/physiology* ; Cohort Studies ; Patient Discharge ; Ischemic Stroke ; Stroke ; Brain Ischemia ; Diabetes Mellitus ; Hypertension

Objective:To explore the status of coronavirus disease 2019 (COVID-19) vaccines, safety and the influencing factors of adverse reactions in maintenance hemodialysis (MHD) patients.Methods:The study was a retrospective study. The MHD patients vaccinated with COVID-19 vaccines in Tianjin city from January 2020 to July 2022 were enrolled in the study. The data of general information, vaccination situation, adverse reactions, and laboratory tests before and after vaccination were collected. Logistic regression analysis was used to analyze the risk factors of adverse reactions after vaccination.Results:A total of 7 375 patients were registered to receive hemodialysis treatment in Tianjin city, of whom 1 036 patients (14.05%) vaccinated with COVID-19 vaccines were enrolled from 53 hemodialysis centers in the study, with age of (54.00±13.27) years old (17-88 years old), and 676 males (65.25%). There were 171 patients (16.51%) receiving the first dose of vaccines only, 464 patients (44.79%) receiving two doses of vaccines, 401 patients (38.71%) receiving three doses of vaccines, and 67 patients (6.47%) had adverse reactions. No serious adverse reaction occurred. The number of neutrophils after vaccination was lower than that before vaccination ( P < 0.05), while the number of lymphocytes, alanine aminotransferase, glutamic oxaloacetic aminotransferase, and serum albumin after vaccination were higher than those before vaccination (all P < 0.05). Logistic regression analysis showed that age ( OR=0.967, 95% CI 0.946-0.990, P=0.005), previous allergic history ( OR=0.013, 95% CI 0.001-0.151, P < 0.001), serum uric acid ( OR=1.004, 95% CI 1.001-1.008, P=0.020), numbers of vaccinations administered ( OR=0.505, 95% CI 0.330-0.774, P=0.002), leukocytes ( OR=0.766, 95% CI 0.628-0.935, P=0.009) and lymphocytes ( OR=0.082, 95% CI 0.045-0.148, P < 0.001) were independently correlated with the incidence of adverse reactions. Conclusions:The proportion of MHD patients vaccinated with COVID-19 vaccines is 14.05%. The incidence of adverse reactions is 6.47%, and there is no serious adverse reaction. Age, previous allergic history, serum uric acid, and numbers of vaccinations administered, leukocytes and lymphocytes are independently correlated with the incidence of adverse reactions in MHD patients.

Objective:To investigate the relationship between microRNA (miR)-1, miR-133b and hepatic fibrosis in patients with hepatic alveolar echinococcosis.Methods:From October 2020 to April 2021, patients who were definitely diagnosed as hepatic alveolar echinococcosis (9 cases), cirrhosis (9 cases) and hepatocellular carcinoma (5 cases) in the First Affiliated Hospital of Xinjiang Medical University were selected as the research subjects, and healthy volunteers in the same period were taken as the control (10 cases). Peripheral blood samples of all subjects were collected to prepare plasma, and the expression levels of miR-1 and miR-133b in peripheral blood were detected by quantitative real-time PCR. At the same time, tissue samples around the liver lesion (proximal), and the corresponding tissues about 5 cm from the lesion (distal) were collected from 5 patients with hepatic alveolar echinococcosis, and immunohistochemical staining was used to detect the cell activation related indicators [cyclinD1, cyclin dependent kinase 1 (CDK1), α-smooth muscle actin (α-SMA)], fibrosis indicators (Collagen Ⅰ, Collagen Ⅲ), transforming growth factor-β1 (TGF-β1) signal pathway related genes [TGF-β1, TGF-β1 receptor type Ⅰ/Ⅱ (TGF-β1RⅠ, TGF-β1RⅡ)] and its downstream related proteins (SMAD2, SMAD3).Results:The quantitative real-time PCR results showed that there were significant differences in the expression levels of miR-1 and miR-133b in the peripheral blood of patients with hepatic alveolar echinococcosis, cirrhosis, hepatocellular carcinoma and the control group ( H = 16.54, 28.40, P < 0.001); the expression levels of miR-1 and miR-133b in hepatic alveolar echinococcosis group were higher than those in control group, cirrhosis group ( P < 0.05). The expression levels of CDK1 (0.46 ± 0.02, 0.42 ± 0.01), α-SMA (0.54 ± 0.09, 0.51 ± 0.07), TGF-β1 (0.55 ± 0.15, 0.51 ± 0.13), TGF-β1RⅠ (0.58 ± 0.09, 0.57 ± 0.08), and TGF-β1RⅡ(0.40 ± 0.05, 0.39 ± 0.05) between the proximal and distal tissue of liver lesion in hepatic alveolar echinococcosis patients were statistically significantly different ( t = 5.56, 3.17, 3.18, 4.27, 5.65, P = 0.005, 0.034, 0.034, 0.024, 0.011). There was no statistically significant difference in the expression levels of CyclinD1, Collagen Ⅰ, Collagen Ⅲ, SMAD2 and SMAD3 between the proximal and distal tissue of liver lesion in hepatic alveolar echinococcosis patients ( t = 3.06, 3.06, 2.86, 1.43, 1.50, P = 0.055, 0.055, 0.064, 0.247, 0.230). Pearson correlation analysis showed that miR-1 in the patients' peripheral blood was positively correlated with TGF-β1RⅠ in the proximal tissue of the liver lesion ( P = 0.001); there was no correlation between miR-1, miR-133b and CDK1, α-SMA, TGF-β1, TGF-β1RⅡ( P > 0.05). Conclusions:The expression of TGF-β1 signaling pathway related factors in the proximal tissue of liver lesion in patients with hepatic alveolar echinococcosis is up-regulated. The expression of miR-1 and miR-133b in peripheral blood is upregulated, and miR-1 is positively correlated with TGF-β1RⅠ level in proximal tissue of liver lesion, suggesting that miR-1 may promote the occurrence of liver fibrosis in hepatic alveolar echinococcosis.

Objective:This study comprehensively analyzed the genomic characterizations of human parainfluenza virus type 3 (HPIV3) strains circulating in six provinces and cities of China (Beijing, Henan, Jilin, Anhui, Gansu, and Shandong) during the period of 2019-2020. The aim was to elucidate the intricate genetic variations and molecular evolutionary trends within the HPIV3 genome.Methods:Based on genotypic differentiation, genetic divergence, and spatial and temporal distribution, 12 representative HPIV3 strains (including 7 of C3a subtype, 2 of C3b subtype and 3 of C3f subtype) were selected from the aforementioned provinces, and the complete genome sequence was successfully obtained by overlapping amplification of fragments using nested RT-PCR. Subsequently, a complete genome database of global representative HPIV3 strains was constructed and analyzed using bioinformatics tools.Results:The length of complete genome of the 12 HPIV3 strains in the present study varied between 15 227 bp and 15 370 bp, the G+ C content ranged from 35.1% to 35.3% and the nucleotide identity intermediated from 97.6% to 99.6%. Compared with the prototype strain (GenBank accession number: NC_001796.2), the nucleotide identity of 12 HPIV3 strains ranged from 94.2% to 94.5%. Analysis of the complete genome of HPIV3 available in China and globally showed that the genomic variation of HPIV3 was mainly shaped by substitution mutations, and no base deletions or gene recombination were observed.Only a six-base insertion (ATTAAA) was found in the F gene’s 3′UTR region of a representative strain originating from Jilin province (CHN/Jilin036/2019/C3b) in this study, and its potential pathogenic significance needs to be further investigated. Amino acid analysis of the encoded proteins revealed that the C3a lineage of HPIV3, widely prevalent both in China and worldwide, exhibits lineage-specific mutation sites in the N, P and L proteins. Furthermore, within the Chinese prevalent C3a strains, a distinctive mutation site (N216S) in L protein was also identified. Notably, specific variant sites have not been found in Chinese C3b and C3f branch strains. Based on the complete genome, the comprehensive evolutionary analysis showed that the time to the most recent common ancestor (tMRCA) of global HPIV3 strains was estimated to 1927 (95% HPD: 1901-1945), with an average molecular evolutionary rate of 5.29 × 10 -4 substitutions/site/year, while the average molecular evolutionary rate of HPIV3 strains in China is 5.24 × 10 -4 substitutions/site/year. In addition, each gene of HPIV3 was subjected to negative selection pressure, with the P, HN and F genes showing the most significant nucleotide variation and higher rates of molecular evolution than the other genes. Conclusions:This study reveals that the complete genome of HPIV3 strains circulating in six provinces and cities of China tend to evolve conservatively. Moreover, substitution emerge as the main driving force for molecular evolution of HPIV3.

Objective:To investigate whether it is by regulating interleukin 1β ( IL-1β) gene expression that androgen receptor (AR) in macrophages affects hyperphosphate-induced vascular smooth muscle cell calcification. Methods:The chromatin immunoprecipitation (ChIP) experiment was used to determine whether AR was bound to the androgen receptor element (ARE) sequence of IL-1β promoter in THP-1 cells. Whether the AR regulated IL-1β gene expression was detected by luciferase assay experiments. AR of THP-1 cells was silenced and transfected by lentivirus with vector or shRNA. Flow cytometry was used to select positive transfected cells THP-1ARsc (control) and THP-1ARsi (AR silencing) with fluorescent markers. Western blotting was used to detect AR protein levels of THP-1ARsc (control) and THP-1ARsi cells (AR silencing in monocytes). Macrophages MФARsc (control) or MФARsi (AR silencing) were induced by 50 ng/ml phorbol ester. Enzyme-linked immunosorbent assay was used to detect IL-1β expression levels of MФARsc or MФARsi conditioned medium. The human aortic smooth muscle cells (HASMC) were cultured in MФARsc or MФARsi conditioned medium with phosphate (2.5 mmol/L final concentration of sodium dihydrogen phosphate), and Alizarin red S staining was used to analyze HASMC calcification degree. Western blotting was used to detect the expression levels of RUNX2 (osteoblast marker) and SM22α (HASMC marker), and neutralization assay was performed to test IL-1β-mediating effect of macrophages AR on HASMC calcification. Results:AR was bound to ARE sequence of IL-1β promoter and regulated IL-1β gene expression. The expression level of IL-1β protein in conditioned medium of MФARsi cells decreased significantly compared to MФARsc cells ( P<0.001). Compared with MФARsc conditioned medium group, HASMC calcium deposition in MФARsi conditioned medium group decreased significantly, RUNX2 protein decreased and SM22α protein increased (all P<0.05). The degree of HASMC calcification in the MФARsi conditioned medium+IgG antibody group decreased than that in the MФARsc conditioned medium+IgG antibody group significantly, and the degree of HASMC calcification in the MФARsc conditioned medium+IL-1β antibody group decreased significantly than that in the MФARsc conditioned medium+IgG antibody group; while the degree of HASMC calcification in the MФARsi conditioned medium+IgG antibody group and MФARsi conditioned medium+IL-1β antibody group decreased than that in the MФARsc conditioned medium+IL-1β antibody group (all P<0.05). Conclusions:Macrophage AR regulates IL-1β expression by binding to ARE sequence within IL-1β promoter, and IL-1β mediates the effect of macrophage AR on hyperphosphate-induced HASMC calcification.

ObjectiveTo study the effect-enhancing and toxicity-reducing activities of astragalus polysaccharide injection （APS） on U14 cervical cancer in model mice receiving X-ray treatment. MethodU14 mouse cervical cancer cells were cultured in vitro and injected into the right forelimb armpit of Kunming mice for constructing a subcutaneous tumor-bearing model of cervical cancer. The tumor-bearing mice were randomly divided into the model group， X-ray intervention（IR， 6 Gy） group， APS （10 mL·kg^-1·d^-1） group， and IR + APS group. Following the observation of the state， body mass， and food intake of mice in each group， the volume of the tumor was measured. The tumor cell cycle and apoptosis were determined by flow cytometry. The protein and mRNA expression levels of apoptosis-related proteins p53， B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， and cleaved cysteine-dependent aspartate-directed protease-3 （Caspase-3） in tumor tissues were assayed by Western blot and real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultThe comparison with the model group showed that mice in the IR group had poor mental status and reduced mobility. The IR group and IR + APS group exhibited reduced food intake and body mass since the 8^th d （P<0.05， P<0.01） and narrowed tumor volume since the 9^th d （P<0.01）. In the IR group， the proportion of cells in the G₁ phase was increased， while the proportion of those in the S phase was decreased （P<0.01）. In the IR + APS group， the proportion of cells in the G₁ phase rose， whereas the proportion of those in the G₂ and S phases cells declined （P<0.05， P<0.01）. The apoptotic rates in both the IR group and IR + APS group were elevated significantly （P<0.01）. Compared with the model group， the IR group and IR + APS group displayed up-regulated cleaved Caspase-3 and Bax protein and mRNA expression in tumor tissues， but down-regulated Bcl-2 and p53 protein and mRNA expression （P<0.05， P<0.01）. Compared with the IR group， the mice in the IR + APS group had better mobility and hair， normal body mass， and increased food intake （P<0.05）. The tumor volume in the IR + APS group was reduced （P<0.05）. The proportion of cells in the G₂ phase was reduced， but the proportion of those in the S phase was raised （P<0.05）. The apoptosis rate was increased （P<0.05）. The apoptosis-related protein Bax protein expression in the tumor tissue was up-regulated， while the protein expression levels of Bcl-2 and p53 were down-regulated （P<0.05， P<0.01）. ConclusionAPS maintains the life state of U14 cervical cancer model mice treated with X-ray and promotes tumor cell apoptosis， thus enhancing the efficiency and reducing toxicity.

Objective:To understand the genetic characteristics of human adenovirus (HAdV) in severe acute respiratory infection (SARI) cases in Luohe city, Henan province, China.Methods:After viral isolation of HAdV-positive specimens identified in SARI cases from October 2017 to February 2019, the Loop2 region of the Hexon gene was amplified and determined to initially identify the virus type. Then, based on the preliminary screening result, the full length of the sequences of the three target genes (Penton base, Hexon and Fiber) of the viral strains were amplified using specific primers for each HAdV type, and phylogenetic and sequence homology analyses were performed with the prototype strains and the representative strains of the corresponding types at home and abroad to identify the types of viral strains and understand their genetic characteristics.Results:A total of 18 viral isolates were obtained from 27 HAdV-positive throat swab specimens from 783 SARI cases in Luohe city, Henan province, and the molecular typing result showed that these strains belonged to species B (HAdV-3, HAdV-7 and HAdV-55), species C (HAdV-1, P1H2F2, Px1/Ps3H5F5, P89H5F5 and HAdV-6) and species E (HAdV-4). Among them, the highest positive detection rate was found for species C HAdV-1 isolates (33.3%), followed by species B HAdV-3 (22.2%) and species C P1H2F2 (11.1%). The four HAdV strains in this study (HAdV-3, HAdV-4, HAdV-7 and HAdV-55) were characterized by significantly conserved and stable in time and space; while three patterns of genetic recombination (P1H2F2, Px1/Ps3H5F5 and P89H5F5) were identified for HAdV-C strains in this study, and their potential public health significance needed to be confirmed by further studies.Conclusions:The HAdV infection of SARI cases in Luohe city, Henan province during 2017-2019 was dominated by species C, followed by species B and species E. These data provided a scientific basis for the prevention and control of local adenovirus-associated infectious diseases.

Objective:To evaluate the effect of synthetic CpG oligodeoxynucleotide (CpG-ODN) as adjuvant on immune response induced by inactivated human adenovirus (HAdV)-55 antigen in BALB/c mice.Methods:HAdV-55 virus QS prototype strain was purified by plaque to construct a seed bank of vaccine candidate strain. The amplified product of vaccine candidate strain was inactivated by 0.05%β-propiolactone, and purified to prepare perfect virus particle antigen. The purified HAdV-55 antigen was mixed with the same volume CPG-ODN and aluminum hydroxide adjuvant in low-dose group (0.2 mg/ml) and high-dose group (1 mg/ml), respectively, and inoculated BALB/c mice after emulsification. Meanwhile, the control group was set with PBS, and the immunization was enhanced once every 21 days. Respectively on primary immune 21 and 35 days after collecting venous blood in mice and separation of serum, serum was collected at the end of the time of separating spleen lymphocytes in mice. The levels of HAdV-55 specific IgG antibody and neutralization antibody in serum of immunized mice were observed by ELISA and micro-neutralization test, and the levels of lymphocytes secreting IL-4 and IFN-γ cytokines were detected by ELISpot.Results:No matter with or without adjuvant, along with the increase of the number of immunization and vaccination dose of inactivated HAdV-55 antigen induced BALB/c mice virus specific IgG antibody was also significantly increased. However, neutralizing antibody can reach detectable level only after enhanced immunity, and the geometric mean titer (GMT) of neutralizing antibody is between 1: 11 and 1: 23. Different adjuvants have significant effects on the immune response of mice. Low dose antigen combined with CPG-ODN and aluminum hydroxide mixed adjuvant can induce higher humoral and cellular immune responses in mice, and the levels of specific IgG antibody and neutralizing antibody are 2.2 and 1.8 times higher than those in the aluminum hydroxide adjuvant group, respectively. The number of lymphocytes secreting IFN-γ was 2.3 times that of the group immunized with aluminum hydroxide adjuvant.Conclusions:The novel CPG-ODN adjuvant significantly increased the immunogenicity of the inactivated HAdV-55 whole virus antigen in BALB/c mice and directed the cellular immune response toward Th1 type.

Objective:To study the effect of shear stress on the expression of KLF2 and eNOS in venous endothelial cells under physiological and uremic conditions, and to explore the mechanism leading to dysfunction of venous endothelial cells.Methods:Under physiological conditions and uremia conditions, different shear stresses were simulated in the parallel plate flow cavity, and the shear stresses were applied to the venous endothelial cells of each group for 4, 12, and 24 hours. The expression of KLF2 and eNOS was detected by immunohistochemical fluorescent staining technique and real-time fluorescent quantitative PCR technique.Results:Under physiological conditions, KLF2 is obviously regulated by shear stresses. High-intensity shear stresses and physiological shear stresses will up-regulate the expression of KLF2, while low-intensity shear stresses and oscillating shear stresses will down-regulate the expression of KLF2. As the duration of action increases, the expression of KLF2 will also increase. In the state of uremia, the expression of KLF2 is significantly inhibited. Even if high shear stresses is applied, the level of KLF2 is not high-expressed as the physiological state. And under the action of low shear stresses and oscillating shear stresses, KLF2 expression is more significantly inhibited. KLF2 is mainly expressed in the nucleus. With the action of shear stresses, KLF2 is also expressed in the cytoplasm, while eNOS is mainly expressed in granular form in the cytoplasm and nucleus.Conclusions:After arteriovenous fistula operation, the expression of KLF2 and eNOS is inhibited under the action of multiple factors of uremia environment and oscillating shear stresses, which may be the main cause of the occurrence and development of venous endothelial cell dysfunction, intimal hyperplasia, and AVF failure.