Objective:To construct an infectious clone of a Chinese HIV-1 CRF01_AE epidemic strain with strong replication capacity，and comprehensively identify its viral phenotype and replication capacity.Methods:Using the CRF01_AE clinical isolate GX2005002，which was previously isolated from whole blood of an HIV-1-infected individual in China by our laboratory，as the parental strain，the full-length genome of the virus（9.7 kb）was divided into 5' half fragment（5.1 kb）and 3' half fragment（4.6 kb）for amplification. The proviral DNA was used as a template to amplify the virus genome，which was then ligated into the eukaryotic expression vector pEASY-T1. The consistency of its sequence with the parental strain sequence was identified through full-length genome sequencing and phylogenetic analysis. The replication capacity，syncytium inducibility，and cell tropism were experimentally identified to determine its phenotypic consistency with the parental strain.Results:An infectious clone of the CRF01_AE strain was successfully constructed，and its genome sequence exhibited high consistency with the sequence of the parental strain. By transfecting target cells，a derivative virus with infectious activity and replication capability was successfully rescued. The derived virus maintained phenotypic characteristics consistent with the parental strain，such as cell tropism and syncytium inducibility.Conclusion:This study successfully constructed an infectious clone of a Chinese HIV-1 CRF01_AE epidemic strain with clear background and distinct phenotype. The genomic sequence and viral phenotypic characteristics of the derived virus are basically consistent with the parental strain，providing strong representation of the original isolate and serving as a powerful tool for research on the correlation between the genetic characteristics，viral phenotype，and pathogenicity of HIV-1 CRF01_AE strains.

Objective:To evaluate the efficacy and safety of nebulized inhalation of recombinant human interferon (IFN) α1b injection in the treatment of respiratory syncytial virus (RSV) associated lower respiratory tract infections (pneumonia and bronchiolitis) in children.Methods:A randomized, double-blind, parallel, placebo-controlled add-on design was used.Children with pneumonia or bronchiolitis aged 2 months to 5 years who tested positive for RSV antigen within 72 hours of onset from 30 clinical trial sites including Beijing Children′s Hospital, Capital Medical University between February 2021 and December 2022 were included in this study and randomly divided into 2 groups at a ratio of 1∶1 based on a stratified-block method.Both groups received basic treatments such as cough control, asthma relieving, expectorant treatment, fever reduction, oxygen therapy, etc.The experimental group received additional nebulized inhalation of IFN α1b injection at a dose of 2.0 μg/(kg·time), twice a day.The control group received nebulized inhalation of placebo twice a day.Clinical efficacy was evaluated based on indicators such as the duration of clinical symptoms and signs, and the Kaplan-Meier method was used to calculate the median and 95% CI of the duration of clinical symptoms and signs.The Log-rank test was used to compared data between groups.Safety was assessed through the incidence of adverse reactions and laboratory tests, and the Chi-square test was used to analyze the difference between groups. Results:There were 123 children in the experimental group and 122 children in the control group.The median durations of all the 5 clinical symptoms and signs [including shortness of breath, wheezing, dyspnea (visible retractions), decreased transcutaneous oxygen saturation, and abnormal mental state] in the experimental group after treatment were slightly shortened than those in the control group [2.7 d(95% CI: 1.9-3.0 d)] vs.[2.9 d(95% CI: 2.6-3.6 d), P=0.027].The improvement in dyspnea (retractions) was especially pronounced in the experimental group, with a relief rate of 50.0% (0, 100%) on the first day of administration[compared with 0 (0, 50.0%) in the control group ( Z=2.002, P=0.025)].The median duration of dyspnea in the experimental group was nearly 1 day shorter than that in the control group [1.0 d(95% CI: 0.7-1.7 d) vs.1.8 d(95% CI: 1.0-2.5 d), P=0.046].There were no significant difference in hospital stay [6.0(5.0, 8.0) d vs.6.5(5.0, 8.0) d, Z=0.675, P=0.500], oxygen therapy duration [32.0(14.0, 96.3) h vs.39.0 (24.0, 83.2) h, Z=0.094, P=0.925], the recovery rate from clinical symptoms during treatment [(105/106, 99.1%) vs.(96/101, 95.0%)], and recurrence rate [(0/106, 0) vs.(2/101, 2.0%)] between the 2 groups (all P>0.05).However, the above-mentioned four indicators in the experimental group showed a trend of clinical benefits.The quantitative virus detection results showed that the RSV viral load in both groups decreased after treatment compared to before treatment.After 2 days of treatment, the decline rate of RSV viral load from the baseline was 0.90 lg copies/(mL·d) in the experimental group and 0.25 lg copies/(mL·d)in the control group, with a statistically significant difference ( P<0.05).Furthermore, there was no statistically significant difference in the incidence of adverse reactions between the 2 groups ( P>0.05).Importantly, no drug-related serious adverse reactions occurred in both groups. Conclusions:The nebulized inhalation therapy of IFN α1b demonstrates efficacy and safety in treating pediatric RSV associated lower respiratory tract infections.It particularly offers outstanding clinical therapeutic value for severe children.

Objective:To evaluate the efficacy and safety of nebulized inhalation of recombinant human interferon (IFN) α1b injection in the treatment of respiratory syncytial virus (RSV) associated lower respiratory tract infections (pneumonia and bronchiolitis) in children.Methods:A randomized, double-blind, parallel, placebo-controlled add-on design was used.Children with pneumonia or bronchiolitis aged 2 months to 5 years who tested positive for RSV antigen within 72 hours of onset from 30 clinical trial sites including Beijing Children′s Hospital, Capital Medical University between February 2021 and December 2022 were included in this study and randomly divided into 2 groups at a ratio of 1∶1 based on a stratified-block method.Both groups received basic treatments such as cough control, asthma relieving, expectorant treatment, fever reduction, oxygen therapy, etc.The experimental group received additional nebulized inhalation of IFN α1b injection at a dose of 2.0 μg/(kg·time), twice a day.The control group received nebulized inhalation of placebo twice a day.Clinical efficacy was evaluated based on indicators such as the duration of clinical symptoms and signs, and the Kaplan-Meier method was used to calculate the median and 95% CI of the duration of clinical symptoms and signs.The Log-rank test was used to compared data between groups.Safety was assessed through the incidence of adverse reactions and laboratory tests, and the Chi-square test was used to analyze the difference between groups. Results:There were 123 children in the experimental group and 122 children in the control group.The median durations of all the 5 clinical symptoms and signs [including shortness of breath, wheezing, dyspnea (visible retractions), decreased transcutaneous oxygen saturation, and abnormal mental state] in the experimental group after treatment were slightly shortened than those in the control group [2.7 d(95% CI: 1.9-3.0 d)] vs.[2.9 d(95% CI: 2.6-3.6 d), P=0.027].The improvement in dyspnea (retractions) was especially pronounced in the experimental group, with a relief rate of 50.0% (0, 100%) on the first day of administration[compared with 0 (0, 50.0%) in the control group ( Z=2.002, P=0.025)].The median duration of dyspnea in the experimental group was nearly 1 day shorter than that in the control group [1.0 d(95% CI: 0.7-1.7 d) vs.1.8 d(95% CI: 1.0-2.5 d), P=0.046].There were no significant difference in hospital stay [6.0(5.0, 8.0) d vs.6.5(5.0, 8.0) d, Z=0.675, P=0.500], oxygen therapy duration [32.0(14.0, 96.3) h vs.39.0 (24.0, 83.2) h, Z=0.094, P=0.925], the recovery rate from clinical symptoms during treatment [(105/106, 99.1%) vs.(96/101, 95.0%)], and recurrence rate [(0/106, 0) vs.(2/101, 2.0%)] between the 2 groups (all P>0.05).However, the above-mentioned four indicators in the experimental group showed a trend of clinical benefits.The quantitative virus detection results showed that the RSV viral load in both groups decreased after treatment compared to before treatment.After 2 days of treatment, the decline rate of RSV viral load from the baseline was 0.90 lg copies/(mL·d) in the experimental group and 0.25 lg copies/(mL·d)in the control group, with a statistically significant difference ( P<0.05).Furthermore, there was no statistically significant difference in the incidence of adverse reactions between the 2 groups ( P>0.05).Importantly, no drug-related serious adverse reactions occurred in both groups. Conclusions:The nebulized inhalation therapy of IFN α1b demonstrates efficacy and safety in treating pediatric RSV associated lower respiratory tract infections.It particularly offers outstanding clinical therapeutic value for severe children.

Objective:To construct an infectious clone of a Chinese HIV-1 CRF01_AE epidemic strain with strong replication capacity，and comprehensively identify its viral phenotype and replication capacity.Methods:Using the CRF01_AE clinical isolate GX2005002，which was previously isolated from whole blood of an HIV-1-infected individual in China by our laboratory，as the parental strain，the full-length genome of the virus（9.7 kb）was divided into 5' half fragment（5.1 kb）and 3' half fragment（4.6 kb）for amplification. The proviral DNA was used as a template to amplify the virus genome，which was then ligated into the eukaryotic expression vector pEASY-T1. The consistency of its sequence with the parental strain sequence was identified through full-length genome sequencing and phylogenetic analysis. The replication capacity，syncytium inducibility，and cell tropism were experimentally identified to determine its phenotypic consistency with the parental strain.Results:An infectious clone of the CRF01_AE strain was successfully constructed，and its genome sequence exhibited high consistency with the sequence of the parental strain. By transfecting target cells，a derivative virus with infectious activity and replication capability was successfully rescued. The derived virus maintained phenotypic characteristics consistent with the parental strain，such as cell tropism and syncytium inducibility.Conclusion:This study successfully constructed an infectious clone of a Chinese HIV-1 CRF01_AE epidemic strain with clear background and distinct phenotype. The genomic sequence and viral phenotypic characteristics of the derived virus are basically consistent with the parental strain，providing strong representation of the original isolate and serving as a powerful tool for research on the correlation between the genetic characteristics，viral phenotype，and pathogenicity of HIV-1 CRF01_AE strains.

Objective:To screen the HIV standard strains with typical biological characteristics of HIV strains circulating in China through the isolation, culture, genotype and phenotype identification of HIV from the whole blood samples of HIV-infected persons, confirm genetic characteristics, traceability, and in line with the Standard Strains of Pathogenic Microorganism-technical Specifications for Establishment of HIV Strains (T/CPMA 027—2023).Methods:Whole blood samples were collected from 48 HIV infected patients. Peripheral blood mononuclear cells (PBMCs) were isolated from the samples and co-cultured with PBMCs isolated from healthy persons′ whole blood samples to isolate and culture HIV from infected persons. We determined concentration of p24 antigen and the virus titer in the culture supernatant. The viral RNA was extracted from the successfully isolated strains, and the gag, pol genes and env C2V3 fragments of the viral genome were amplified and sequenced. The genotype, gene recombination and drug resistance sites were determined according to the viral gene sequences. Virus infection and replication were monitored by inoculating the virus culture supernatant into Ghost cells expressing CCR5 or CXCR4 to determine the viral tropism.The formation of syncytium was observed by inoculating the virus culture supernatant into MT-2 cells to determine whether was a syncytium-induced phenotype. Results:Fourteen strains with p24 antigen concentration > 1 ng/ml in culture supernatant were isolated and cultured from 48 fresh EDTA anticoagulated whole blood samples of HIV infected persons. Of the 14 strains, only one strain with a titer≥10 5 TCID 50/ml, 8 strains with titers ≥10 4 TCID 50/ml, and the other 5 strains with titers≥10 3 TCID 50/ml. Phylogenetic analysis showed that the genotypes of the strains were 9 strains of subtype B, 3 strains of CRF01_AE and 2 strains of CRF07_BC recombinant. Genotypic resistance analysis showed that 11 strains contained drug resistance sites. Ghost cells were used to verify the tropism of the strains, and it was found that 8 strains were CCR5 tropism, 6 strains were CXCR4 & CCR5 dual tropism. Only 2 of the 14 strains could induce MT-2 cytopathic effect, which was syncytium-inducing phenotype. Conclusions:Fourteen HIV strains with typical biological and genetic characteristics were isolated to screen the standard HIV strains. Among which, 1 strain was evaluated as a standard HIV strain that meets the Standard Strains of Pathogenic Microorganism-technical Specifications for Establishment of HIV Strains (T/CPMA 027—2023). This study can also provide technical guidance for the screening of the HIV standard strains. Next step is to complete the application and reserve database construction according to the sharing mechanism of the HIV standard strains, to provide resources for the researches of HIV vaccines and drugs.

Objective:To analyze the characteristics of elderly patients with moyamoya disease and clinical outcomes after combined revascularization, and to evaluate the safety and efficacy of this method in elderly patients.Methods:Moyamoya disease patients aged 60 years or older who had undergone combined revascularization at the Neurosurgery Center of Aeronautical General Hospital from January 2016 to September 2021 were selected as study subjects.Then patients' clinical data and postoperative complications within two weeks of surgery were collected and a followed-up was conducted.The results were compared with those of previous reports.Results:A total of 46 combined revascularization procedures were carried out in 30 patients.The ages at surgery ranged between 60-75 years, with a mean age of(66.1±4.4)years.The rate of complications within two weeks after surgery was 37%, and the most common complication was speech dysfunction, followed by epilepsy and hyperperfusion.No serious complications such as cerebral hemorrhage and death occurred.The follow-up varied between 3-67 months, with an average follow-up time of(26.8±18.4)months.The MRS score was <3 at 3 months after surgery, and there was no death or severe disability.Conclusions:Perioperative complications in elderly patients with moyamoya disease have a higher incidence rate than in other populations, but patients can usually recover after close perioperative management for 2 weeks.The follow-up has demonstrated that outcomes at 3 months and in longer terms after surgery are satisfactory and clinical symptoms will improve significantly.

Objective:To investigate the predictive neutrophil/lymphocyte ratio (NLR) combined with soluble growth stimulating expression gene 2 protein (sST2) on in-hospital major adverse cardiovascular events (MACE) in patients with myocardial injury following moderate-severe acute carbon monoxide poisoning (ACOP).Methods:A single-cente prospective observational approach was conducted. Moderate-severe ACOP patients with myocardial damage from November 2016 to February 2020 in department of emergency medicine of Harrison International Peace Hospital Affiliated to Hebei Medical University were enrolled. The baseline data of the patients, NLR and sST2 (T0 sST2) on admission, sST2 at 3 days after admission (T3 d sST2), and the other myocardial injury and biochemical indicators were collected. According to whether MACE occurred, the patients were divided into MACE group and non-MACE group. The clinical data of the two groups were compared. Pearson correlation analysis was used to analyze the correlation of each index. Binary Logistic regression was used to analyze the independent risk factors of in-hospital MACE in patients with moderate-severe ACOP myocardial injury. The receiver operator characteristic curve (ROC curve) was drawn and area under ROC curve (AUC) was calculated to analyze the predictive value of NLR, sST2, and NLR combined with sST2 for the occurrence of in-hospital MACE in patients with moderate-severe ACOP myocardial injury.Results:A total of 278 patients with moderate-severe ACOP myocardial injury were included in the final analysis, and the incidence of MACE was 11.51% (32/278). Cardiac troponin I (cTnI), lactic acid (Lac), NLR, and T3 d sST2 in the MACE group were significantly higher than those in the non-MACE group [cTnI (μg/L): 0.83±0.15 vs. 0.46±0.37, Lac (mmol/L): 2.96±1.14 vs. 2.43±1.35, NLR: 13.14±4.37 vs. 9.49±4.21, T3 d sST2 (μg/L): 59.88±23.42 vs. 39.83±12.60, all P < 0.05], there was no significant difference in T0 sST2 between the MACE group and the non-MACE group (μg/L: 269.09±90.89 vs. 240.14±113.02, P > 0.05). Pearson correlation analysis showed that there were significantly positive correlations in NLR with acute physiology and chronic health evaluationⅡ(APACHEⅡ), T3 d sST2 with APACHEⅡ, and NLR with T3 d sST2 ( r values were 0.226, 0.209, 0.193, all P < 0.01). Binary Logistic regression analysis showed that T3 d sST2 and NLR were both independent risk factors for MACE in moderate-severe ACOP patients with myocardial injury [odds ratio ( OR) and 95% confidence interval (95% CI) respectively was 1.064 (1.039-1.090), 1.176 (1.066-1.298), both P < 0.01]. ROC curve analysis showed that the predictive efficacy of NLR combined with T3 d sST2 for the occurrence of in-hospital MACE in patients with ACOP myocardial injury (AUC = 0.876) was better than that of NLR (AUC = 0.754) and T3 d sST2 (AUC = 0.813). When the optimal critical value of NLR was 10.02 and that of T3 d sST2 was 43.50 μg/L, the sensitivity of predicting the occurrence of MACE in patients with moderate-severe ACOP myocardial injury was 69.8% and 86.2% respectively, and the specificity was 74.3% and 70.4%, respectively. The specificity and sensitivity of the combined detection was 83.4% and 79.8%, respectively. Conclusions:NLR and T3 d sST2 were independent predictors of in-hospital MACE in moderate-severe ACOP patients with myocardial injury, and combined application of NLR and T3 d sST2 had good predictive value. For patients with moderate-severe ACOP myocardial injury with NLR > 10.02 and T3 d sST2 > 43.50 μg/L, the occurrence of in-hospital MACE should be alert.

Objective:To investigate the clinical significance of soluble growth stimulating expression gene 2 protein (sST2) combined with neutrophil/lymphocyte ratio (NLR) in the prediction of nosocomial cardiovascular adverse events in patients with acute carbon monoxide poisoning (ACOP) myocardial injury.Methods:Patients with ACOP myocardial injury from January 2017 to December 2019 in Emergency Ward and EICU of Harrson International Peace Hospital, Hebei Medical University were enrolled. NLR was calculated by routine blood examination on admission, and sST2 (T 0sST2, T 3dsST2) was detected by ELISA on admission and at 3 days after admission. According to the occurrence of cardiovascular adverse events, the patients were divided into the event group and the non-event group. Logistic regression was used to analyze the risk factors of in-hospital cardiovascular adverse events. ROC curve was used to analyze the value of sST2, NLR, sST2 and NLR combined in predicting the occurrence of in-hospital cardiovascular adverse events in patients with ACOP myocardial injury. Results:Totally 255 patients with ACOP myocardial injury were included in the final analysis. NLR was (13.38±4.33) in the event group and (9.57±4.22) in the non-event group, T 3dsST2 was (61.59±22.67) ng/mL in the event group and (40.52±13.14) ng/mL in the non-event group, with statistically significant differences (all P<0.01). T 0sST2 was (265.34±89.95) ng/mL in the event group and (242.43±93.09) ng/mL in the non-event group, with no statistically significant difference ( P=0.333). Logistic regression analysis showed that NLR ( OR=1.270, 95% CI: 1.125-1.434, P<0.01) and T 3dsST2 ( OR=1.082, 95% CI: 1.052-1.114, P<0.01) were independent risk factors for nosocomial cardiovascular adverse events in patients with ACOP myocardial injury. The optimal cutoff value of T 3dsST2 was 44.5 ng/mL, and of NLR was 12.08. The sensitivity and specificity of dual T 3dsST2 and NLR in predicting nosocomial cardiovascular adverse events was 79.3% and 82.7%, respectively (AUC 0.857, Youden index 0.620). Conclusions:T 3dsST2 and NLR are independent risk factors for the nosocomial cardiovascular adverse events in patients with ACOP myocardial injury. The predictive cutoff values are 44.5 ng/mL for T 3dsST2 and 12.08 for NLR. Combination of T 3dsST2 and NLR has a practical predictive value for nosocomial cardiovascular adverse events in patients with ACOP myocardial injury.

Objective To investigate the effects of three-dimensional titanium mesh repair on cerebral perfusion,intracranial pressure and nerve function after decompression with bone flap.Methods From October 2015 to October 2018,132 hypertensive cerebral hemorrhage patients of bone disc decompression in the Fifth People's Hospital of Jinan were selected and divided into observation group (66 cases) and control group(66 cases) by double blind random method.The observation group received three-dimensional titanium mesh repair at 24 ～ 42 d after bone disc decompression surgery.The control group received three-dimensional titanium mesh repair at 90 ～ 150 d(3 ～ 5 months) after surgery.The changes of operation,cerebral perfusion,intracranial pressure and nerve function were compared between the two groups.Results The operative time,blood loss and free time of the flap in the observation group were (92.5 ± 12.4) min,(354.3 ± 17.5) mL and (13.2 ± 3.1) min,respectively,which were shorter or less than those in the control group [(142.8 ± 15.3) min,(518.3 ± 22.3) mL and (38.3 ± 4.3) min],the differences between the two groups were statistically significant (t=4.745,6.831,4.963,all P ＜ 0.05).After treatment,the abnormal indices of intracranial pressure and cerebral perfusion in the observation group were (0.1 ±0.0) and(0.2 ± 0.1),respectively,which were lower than those in the control group [(0.2 ± 0.1),(0.3 ± 0.1)] (t =3.657,2.579,all P ＜0.05).There were statistically significant differences in neurological deficit scores between the two groups at different time points(all P ＜0.05).The incidence of treatment complications in the observation group was 10.94％ (7/64),which was significantly lower than that in the control group [38.7％ (24/62)] (x2 =14.094,P ＜ 0.05).Conclusion Three-dimensional titanium mesh repair is helpful to shorten the operation time,restore the intracranial pressure on the affected side,improve the abnormal cerebral perfusion,and reduce the neurological damage in patients with hypertensive cerebral hemorrhage.

Objective To investigate the clinical effect of paraquat (PQ) detoxification recipe combined with continuous hemoperfusion (HP) in the treatment of patients with acute paraquat poisoning (APP) and clinical significance of soluble CD14 subtype (sCD14-st, Presepsin). Methods A prospective randomized controlled trial was conducted. 152 patients with moderate APP admitted to Department of Emergency Medicine of Harrison International Peace Hospital Affiliated to Hebei Medical University from July 2013 to June 2017 were enrolled, and they were randomly divided into three groups. The patients in HP group (group A, n = 35) only received 2-hour HP for 3 times, 8 hours each time, those in PQ detoxification recipe combined with HP group (group B, n = 50) received PQ detoxification recipe 1 (once per 2 hours until no PQ component was found in faeces) and 2 (3 times a day for 14 days) beside HP. The others in PQ detoxification recipe combined with persistent HP group (group C, n = 67) received continuous HP until the PQ component in serum was not detected. The parameters of organ function and inflammatory factor, and blood Presepsin and PQ contents were determined before and after treatment. The curative effect and 28-day mortality were recorded. The correlations between serum Presepsin level and PQ content as well as 28-day mortality were analyzed with Pearson correlation analysis. Receiver operating characteristic curve (ROC) was plotted to analyze the predictive value of Presepsin on prognosis. Results The total effective rate of group C was significantly higher than that of groups A and B [70.1% (47/67) vs. 34.3% (12/35), 54.0% (27/50)], and 28-day mortality was significantly lowered [29.8% (20/67) vs. 65.7% (23/35), 46.0% (23/50), both P < 0.05]. There was no significant difference in alanine aminotransferase (ALT), MB isoenzyme of creatine kinase (CK-MB), serum creatinine (SCr), C-reactive protein (CRP), tumor necrosis factor-α (TNF-α), and interleukins (IL-6 and IL-10) before treatment among the three groups. Five days after treatment, the above parameters in the three groups were increased as compared with those before treatment, but the increase degree in group C was the lowest. At 7 days after treatment, the parameters were decreased, especially in group C. There was no significant difference in serum Presepsin and PQ levels before treatment among the three groups. With the prolongation of treatment time, the Prespsin levels in groups A, B, and C were increased, and peaked at 12 hours (μg/L: 4.28±0.20, 3.87±0.25, 3.53±0.23), then gradually decreased,and the PQ contents were lower than those before treatment from 8 hours (mg/L: 1.76±0.12 vs. 2.12±0.17, 1.57±0.08 vs. 2.24±0.16, 1.25±0.10 vs. 2.14±0.18), with a time dependence pattern, especially in group C (all P < 0.05) . Correlation analysis showed that blood Presepsin level was positively correlated with PQ content and 28-day mortality (r1= 0.917, r2= 0.864, both P = 0.001), suggesting that the higher the PQ content was, the higher the Presepsin level, and the higher the 28-day mortality was. ROC curve analysis showed that the area under ROC curve (AUC) of Presepsin predicting 28-day mortality was 0.863; when the cut-off value was 1.22 μg/L, the sensitivity was 83.3%, the specificity was 81.4%, the positive predictive value was 77.46%, and the negative predictive value was 86.42%. Conclusions Early administration of PQ detoxification recipe combined with continuous HP treatment can effectively reduce Presepsin level, decrease the mortality of patients with moderate APP, improve the prognosis. Presepsin can assess the prognosis of patients with APP.