Objective To detect the serum levels of CC chemokine receptor 2(CCR2)and C-reactive pro-tein(CRP)in stroke patients,and analyze their relationship with the severity of stroke associated pneumonia and their clinical significance.Methods A total of 78 patients with stroke associated pneumonia who were di-agnosed and treated in the hospital from October 2022 to February 2023 were collected as the study group,ac-cording to the severity of pneumonia,the study group was divided into mild group(31 cases),moderate group(29 cases),and severe group(18 cases),78 stroke patients who did not develop pneumonia were included into control group.Pearson method was applied to analyze the correlation between serum CCR2 and CRP levels in stroke associated pneumonia patients.Multivariate Logistic regression was applied to analyze the factors influ-encing the occurrence of stroke associated pneumonia.Receiver operating characteristic(ROC)curve was ap-plied to analyze the diagnostic value of serum CCR2 and CRP for stroke associated pneumonia.Results The National Institute of Health stroke scale(NIHSS)score,serum CCR2,and CRP levels in the study group were obviously higher than those in the control group(P<0.05).The levels of serum CCR2 and CRP increased with the aggravation of pneumonia(P<0.05).The levels of serum CCR2 and CRP in the study group were positively correlated(r=0.799,P<0.05).NIHSS score,CCR2,and CRP levels were risk factors for stroke associated pneumonia in stroke patients(P<0.05).The area under the curve(AUC)for the diagnosis of stroke associated pneumonia using serum CCR2 and CRP alone was 0.873 and 0.888,respectively,and the AUC for the combined detection of the two was 0.936,the combined detection of the two was superior to the individual detection of serum CCR2 and CRP(Zcombination-CCR2=1.987,Zcombination-CRP=1.832,P=0.041,0.047).Conclusion Serum CCR2 and CRP are closely related to the severity of stroke associated pneumonia,and their combined detection has high diagnostic value for stroke associated pneumonia.

Objective To investigate the diagnostic value of serum soluble fms-like tyrosine kinase 1(sFlt-1)and soluble differentiation cluster 14(sCD14)in stroke-associated pneumonia(SAP).Methods A total of 67 SAP patients admitted to the hospital from March 2022 to January 2023 were selected as the study group,and 50 stroke patients without pneumonia during the same period were selected as the control group.Enzyme-linked immunosorbent assay was used to measure serum sFlt-1 and sCD14 levels.Pearson correlation analysis was used to analyze the correlation between serum sFlt-1,sCD14 levels and clinical data.Multivariate Logistic regression was used to analyze the influencing factors of SAP in patients with stroke.Receiver operating char-acteristic(ROC)curve was used to analyze the diagnostic value of serum sFlt-1 and sCD14 levels in predicting SAP in patients with stroke.Results The serum levels of sFlt-1 and sCD14 in SAP patients were higher than those in stroke patients without pneumonia and stroke patients with pneumonia alone(P＜0.05).SAP in stroke patients was associated with age,atrial fibrillation,heart failure,aspiration,dysphagia,brain stem stroke,proton pump inhibitor use,fever,cough,dyspnea,low-density lipoprotein cholesterol,procalcitonin(PCT),C-reactive protein,white blood cell count,national institutes of health stroke scale(NIHSS)score,clinical pulmonary infection(CPIS)score(P＜0.05).Correlation analysis showed that serum sFlt-1 level was positively correlated with sCD14(r=0.439,P＜0.001),and serum sFlt-1 and sCD14 levels were positively correlated with NIHSS score and CPIS score(P＜0.05).Multivariate Logistic regression analysis showed that sFlt-1,sCD14,proton pump inhibitor use,PCT,dysphagia,and age were the influencing factors of SAP in stroke patients(P＜0.05).ROC curve results showed that the area under the curve of serum sFlt-1 and sCD14 combined to evaluate SAP in stroke patients was higher than that of single detection(ZsFlt-1-combined=2.194,P=0.028,ZscD14-combined=2.310,P=0.002).Conclusion The serum levels of sFlt-1 and sCD14 are in-creased in SAP patients,and the combination of the two has a good diagnostic value for predicting the occur-rence of SAP.

Objective To investigate the effect of different doses of esketamine combined with hydromorphone postoperative patient-controlled intravenous analgesia(PCIA)on depression in elderly pa-tients undergoing total knee arthroplasty.Methods A total of 180 elderly patients,44 males and 136 fe-males,aged 65-80 years,BMI 18.5-35.0 kg/m2,ASA physical status Ⅱ or Ⅲ,undergoing total knee arthroplasty(TKA)under elective general anesthesia combined with adductor block from J uly 2023 to Sep-tember 2023.Patients were divided into three groups by random number table method;control group(group C),esketamine 0.5 mg/kg group(group E1),and esketamine 1.0 mg/kg group(group E2),60 patients in each group.After operation,groups C,E1 and E2 were given hydromorphone 0.2 mg/kg,esketamine 0.5 mg/kg combined with hydromorphone 0.2 mg/kg,and esketamine 1.0 mg/kg combined with hydromor-phone 0.2 mg/kg to receive PCIA,respectively,and the three groups were diluted to 100 ml with normal saline.Parameters were set as follows.The background infusion rate was 1.5 ml/h,and the single press dose was 1.5 ml,and the locking time was 15 minutes.If the VAS pain score at rest was greater than or equal to 4 points and the analgesic effect of pressing the PCIA pump was not effective,then intramuscular injection of tramadol 0.1 g was used for remedial analgesia.Hamilton depression scale(HAMD)score was performed 1 day,3 and 7 days after surgery.Depressive state was classified as having HAMD score ≥ 8 points.VAS pain scores at rest were performed 1 day,3 and 7 days after surgery.The number of depression within 7 days after surgery,the number of effective(D1)and total(D2)pump compressions and D1/D2 within 3 days after surgery,the number of rescue analgesia,the occurrence of adverse reactions such as tra-madol dosage,dizziness,headache,multiple dreams,hallucinations,nausea and vomiting were recorded.Results Twenty-one patients(35％)in group C experienced depression,7 patients(12％)in group E1,and 8 patients(13％)in group E2 during 3 days after surgery.Eight patients(13％)in group C experi-enced depression,1 patients(2％)in group E1,and 2 patients(3％)in group E2 during 7 days after sur-gery.Compared with group C,the incidence of depression 3 and 7 days after surgery,rescue analgesia rate in group E1 were significantly decreased,the incidence of depression 3 and 7 days after surgery,dizziness,headache,and dreaminess within 3 days after surgery in group E2 were significantly decreased(P＜0.05).There were no significant differences in the incidence of depression and VAS pain scores between group El and group E2 at 1,3,and 7 days after surgery.Conclusion Esketamine 0.5 and 1.0 mg/kg for PCIA in elderly patients after TKA can improve postoperative depression,while esketamine 1.0 mg/kg can reduce the incidence of postoperative dizziness,headache,and multiple dreams.

Objective:To evaluate the clinical efficacy of omalizumab in the treatment of patients with chronic spontaneous urticaria accompanied by other allergic diseases.Methods:Clinical data were retrospectively collected from 74 patients, who were clinically diagnosed with chronic spontaneous urticaria and other allergic diseases, and received subcutaneous injections of omalizumab in the Department of Allergy, Tianjin Medical University General Hospital from June 2020 to September 2022. Types of allergic diseases, serum total IgE (tIgE) and allergen-specific IgE (sIgE) levels before treatment, treatment outcomes and adverse drug reactions were analyzed. Differences before and after treatment were assessed using paired t-test and Wilcoxon signed-rank sum test. Results:A total of 74 patients with chronic spontaneous urticaria were involved, including 29 with complicated allergic asthma (39.2%) , 61 with complicated allergic rhinitis (82.4%) , 6 with complicated atopic dermatitis (8.1%) , and 4 with food allergy (5.4%) . Before treatment, elevated serum tIgE or sIgE levels were observed in 44 (59.5%) patients. After the first omalizumab treatment, the urticaria control test (UCT) score significantly increased compared with that before treatment (16.00 [13.0.0, 16.00] vs. 6.00 [5.75, 9.00], Z = 7.39, P < 0.001) ; after 4 sessions of the omalizumab treatment, 82.5% (33/40) of the patients achieved complete control of urticaria symptoms or showed complete response. After omalizumab treatment, asthmatic attacks were decreased in 29 patients with allergic asthma, and asthma control test (ACT) scores significantly increased compared with those before treatment (21.07 ± 2.88 points [after the first treatment] vs. 18.48 ± 3.20 points [before treatment], t = 8.87, P < 0.001) ; among 61 patients with allergic rhinitis, global rhinitis symptom-based visual analog scale (VAS) scores (before treatment: 5.89 ± 1.29 points; after the first treatment: 3.28 ±1.46 points) and rhinoconjunctivitis quality of life questionnaire (RQLQ) scores (before treatment: 60.10 ± 20.53 points; after the first treatment: 37.26 ± 18.83 points) both significantly decreased after the first treatment ( t = 15.04, 10.01, respectively, both P < 0.001) , and rhinitis symptoms were relieved at the same time; skin itching was relieved in 4 patients with atopic dermatitis, and allergic symptoms after contact with food allergens were also relieved in the 2 patients with food allergy after omalizumab treatment. During the treatment, only 1 patient experienced erythematous swelling, induration, and pain at the injection site. Conclusions:In the treatment of chronic spontaneous urticaria accompanied by allergic diseases, the use of omalizumab not only effectively improved urticaria symptoms, but also well controlled allergic diseases, with a good safety profile. Multiple benefits may be achieved by the use of omalizumabin in patients with chronic spontaneous urticaria accompanied by other allergic diseases.

Objective:To preliminarily evaluate the immunogenicity and efficacy of two novel tuberculosis vaccine candidates (a fusion multicomponent protein EPDPA015f and a mixed multicomponent protein EPDPA015m) and to provide a new antigen combination for the development of tuberculosis vaccines.Methods:Recombinant plasmids for the expression of EPDPA015f and EPDPA015m proteins were constructed. Six-week-old BALB/c mice were immunized with EPDPA015f or EPDPA015m in combination with aluminium adjuvant (50 μg/mouse) for three times with an interval of 10 d. The mice were sacrificed 10 d after the last immunization to collect blood and spleen samples. Serum antibody titers and cytokine levels were measured by ELISA, Luminex technique and enzyme-linked immunospot assay (ELISPOT). Mycobacterial growth inhibition assay (MGIA) was used to detect the ability of mouse splenocytes to inhibit the growth of Mtb in vitro. One-way analysis of variance and t-test were used for statistical analysis. Results:Both EPDPA015f and EPDPA015m could induce the production of various cytokines and IgG antibodies at a high level. The levels of cytokines related to Th1 (IL-2, TNF-α, IFN-γ), Th2 (IL-4, IL-6, IL-10) and Th17 (IL-17) as well as other proinflammatory cytokines (GM-CSF, IL-12) were higher in the EPDPA015f group than in the adjuvant group ( P<0.05). The titer of IgG antibody induced by EPDPA015f was as high as 1∶4×10 6. The results of MGIA showed that the numbers of Mtb (lgCFU) in the PBS, adjuvant, EPDPA015f and EPDPA015m groups were 3.46±0.11, 3.51±0.06, 2.98±0.09 and 3.19±0.08, respectively. The number of colonies in the EPDPA015f group was the least as compared with that in the other three groups ( P<0.001, P<0.001, P<0.01). Conclusions:The vaccine candidate EPDPA015f could elicit more comprehensive and high-level cellular and humoral immune responses, and exhibited superior in vitro inhibitory activity against the growth of Mtb. EPDPA015f had the potential to be used as a preventive vaccine or a booster vaccine

OBJECTIVE: To explore the pathogenic variants and clinical classification of two fetuses with Short-rib thoracic dysplasia with or without polydactyly (SRTD). METHODS: With informed consent obtained, the phenotypic characteristics of the fetuses were comprehensively examined, and genomic DNA was extracted from fetal skin tissue and peripheral blood samples of the parents with conventional phenol-chloroform method. Whole exome sequencing (WES) was carried out on both fetuses, and the candidate variants were validated by Sanger sequencing. The pathogenicity of the candidate variants was analyzed using bioinformatic software VarCards, and the impact of the variants on the protein structure was predicted with Swiss-Pdb-viewer. RESULTS: Both fetuses were found to harbor compound heterozygous variants of the DYNC2H1 gene, including c.515C>A (p.Pro172Gln) and c.5983G>A (p.Ala1995Thr) in fetus 1, and c.5920G>T (pGly1974) and c.9908T>C (p.He3303Thr) in fetus 2. The parents of both fetuses were heterozygous carriers. CONCLUSION The compound heterozygous variants of the DYNC2H1 gene probably underlay the SRTD3 in the two fetuses.
Humans ; Fetus ; Chloroform ; Computational Biology ; Ethnicity ; Ribs

Objective:To evaluate the immunogenicity and protective effect of a multicomponent recombinant protein vaccine EPRHP014 constructed independently and provide a scientific basis for developing new tuberculosis (TB) vaccine and effective prevention and control of TB.Methods:Three full-length Mycobacterium ( M.) tuberculosis protein antigens (EsxH, Rv2628, and HspX) and two epitope-predicted and optimized epitope-dominant protein antigens (nPPE18 and nPstS1) were selected, from which five protein antigens were used to construct a protein antigen composition EPRHP014, including a fusion expression multi-component protein antigen (EPRHP014f) and a multi-component mixed protein antigen (EPRHP014m) formed with the five single protein using clone, purification, and purification respectively. Multicomponent protein vaccines EPRHP014f and EPRHP014m were prepared with aluminum adjuvant, and the BCG vaccine was used as a control. ELISA detected the titer of serum-specific antibodies, the secretion of various cytokines was detected by ELISpot and Luminex, and immune protection was observed by the M.tuberculosis growth inhibition test in vitro. The results were statistically analyzed by t-test or rank sum test, and P<0.05 was considered a statistically significant difference. Results:Mice Immunized with EPRHP014m and EPRHP014f could produce highly effective IgG antibodies and their subtypes IgG1 and IgG2a, and the antibody titers were similar to those of mice immunized with BCG, with no statistical significance ( P>0.05). The number of spot-forming cells (SFC) secreting IFN-γ and IL-4 induced by EPRHP014f group was significantly higher than those by EPRHP014m group and BCG group ( P<0.05), but there was no significant difference in the number of SFC for IFN-γ and IL-4 induced between EPRHP014m group and BCG group ( P>0.05). The secretion levels of GM-CSF and IL-12p70 induced by the EPRHP014m group were higher than those of the BCG group ( P<0.05), but there was no significant difference in the levels of IL-6 and IL-10 induced between EPRHP014m group and BCG group ( P>0.05). There was no significant difference in the secretions of IL-6, IL-10, IL-12, and GM-CSF between the EPRHP014f and BCG groups ( P>0.05). EPRHP014m group, EPRHP014f group, and BCG group had obvious antibacterial effects in vitro, and the difference was insignificant ( P>0.05). Conclusion:Both EPRHP014f and EPRHP014m can induce strong humoral and cellular immune responses in mice after immunization, and have a strong ability to inhibit the growth of M. tuberculosis in vitro, indicating that the antigen composition EPRHP014 has good potential in the development and application of TB vaccine.

OBJECTIVE To analyze quality maker （Q-marker） of Ka nggongyan soft capsule （KSC）. METHODS The fingerprints of 20 batches of KSC were established by ultra high performance liquid chromatography （UPLC）method. Similarity Evaluation System of TCM Chromatographic Fingerprint （2012 edition）were used to evaluate the similarity and confirm common peaks. The contents of norisoboldine ，leonurine hydrochloride ，forsythoside B ，acteoside，poliumoside and isoacteoside were determined by the same UPLC method. Targets and pathways related to KSC in the treatment of cervicitis were screened and analyzed by network pharmacology and molecular docking method to construct a “component-target-pathway”network，and analyze its potential Q-marker. RESULTS Twelve common peaks were identified in the fingerprints of 20 batches of KSC ，and the similarity was greater than 0.99. Six common peaks were identified ，including norisoboldine ，leonurine hydrochloride ，forsythoside B，acteoside，poliumoside and isoacteoside. The contents of the above 6 components were 1.336-1.774，0.093-0.143，4.970-5.888， 0.505-0.623，5.206-6.226 and 0.785-0.895 mg/g，respectively. By network pharmacology analysis ，14 key targets and 94 pathways were obtained ，and their binding energies to the core targets （protein kinase B 1，tumor necrosis factor ）were all less than －6.4 kJ/cal. CONCLUSIONS Six components such as norisoboldine and leonurine hydrochloride are potential Q-marker of KSC.

Objective:To explore the guiding effect of peripheral perfusion index (PI) on fluid resuscitation in patients with septic shock.Methods:Sixty-five patients with septic shock who were diagnosed according to relevant criteria of septic shock and admitted to the department of critical care medicine of the Affiliated Hospital of Hangzhou Normal University from September 2017 to December 2020 were included. Patients were divided into the conventional treatment group (30 cases) and PI guidance group (35 cases) by random number method. Both groups of patients were treated with the bundle according to clinical guidelines. Sputum, urine and blood were collected for pathogenic microorganism culture before the application of antibiotics, and vasoactive drugs were given. Both groups need to achieve all the following resuscitation goals within 6 hours: urine output > 0.5 mL·kg -1·h -1, mean arterial pressure (MAP) ≥ 65 mmHg (1 mmHg ≈ 0.133 kPa), central venous pressure (CVP) was 8-12 mmHg, and central venous oxygen saturation (ScvO 2) ≥ 0.70. There was no further resuscitation in the conventional treatment group after the goals were achieved. In addition to these four goals, the PI guidance group was expected to achieve PI≥ 1.4. Heart rate (HR), CVP, MAP, ScvO 2, blood lactic acid (Lac), the time of fluid negative balance, intensive care unit (ICU) mortality and 28-day mortality between the two groups were compared before and after 6 hours of fluid resuscitation. Results:Before fluid resuscitation, there were no statistically significant differences in all indicators between two groups. After 6 hours fluid resuscitation, the four treatment goals in PI guidance group were slightly lower than those of the conventional treatment group [HR (times/min): 96.5±12.1 vs. 97.7±7.9, MAP (mmHg): 83.2±6.2 vs. 82.1±7.5, ScvO 2: 0.661±0.077 vs. 0.649±0.051, CVP (mmHg): 10.8±2.7 vs. 10.4±2.1], there were no statistically significant differences between the two groups (all P > 0.05); the Lac level of the PI guidance group after resuscitation was lower than that of the conventional treatment group, and the difference was statistically significant (mmol/L: 4.8±1.3 vs. 5.9±1.4, P < 0.05); the duration of fluid negative balance in the PI guidance group was earlier than that in the conventional treatment group [days: 3.0 (2.0, 3.0) vs. 3.5 (3.0, 4.0), P < 0.05]. The ICU mortality and 28-day mortality in the PI guidance group were lower than those in the conventional treatment group [ICU mortality rate: 37.1% (13/35) vs. 50.0% (15/30), 28-day mortality rate: 57.1% (20/35) vs. 60.0% (18/30)], but the differences were not statistically significant (both P > 0.05). Conclusions:The peripheral PI can be used as an important indicator of fluid resuscitation in patients with septic shock. PI guiding fluid resuscitation in patients with septic shock can reduce Lac levels, shorten the duration of fluid negative balance and reduce the risk of fluid overload.

Objective:To analyze the change trend of survival rate after hip fragility fracture in Tianjin Hospital from 2015 to 2021, and the influence of gender, marital status, age and number of complications on survival rate.Methods:From January 1, 2015 to December 31, 2021, a total of 12,570 patients with fragility fracture of hip were retrieved, including 3,934 males and 8,636 females. The age at admission was 74.11±9.50 years and 74.62±8.99 years respectively. By comparing the ID number with the Tianjin population information database, 2,054 cases died, including 804 males and 1,250 females, aged 81.34±7.88 years and 81.92±7.42 years respectively at the time of death. Acquire the patient's survival status, calculate the cumulative survival rate at 3 month intervals, study the change rule of the cumulative survival rate over time, and use Kaplan-Meier method to calculate the cumulative survival rate of the whole population and the impact of gender, marital status, age, and number of complications on the survival rate.Results:The median survival time of all the dead people after fracture was 13(3, 31) months, including 11(2, 27) months for males and 15(4, 32) months for females. Joinpoint regression showed that 9 months after the hip fragility fracture was the break point of the survival rate. The mortality rate changed significantly within 9 months after fracture (the annual change rate was 47%), and slowed down 9 months later (the annual change rate was 1%). There was a statistically significant difference in trend detection before and after the break point ( P<0.05). The age at admission was 80.11±7.71 years for the dead and 73.36±9.01 years for the non dead, with a statistically significant difference ( t=31.80, P<0.001). After normalization, the number of complications was 0.20±0.93 among the dead and 0.00±0.87 among the non dead, with a statistically significant difference ( t=88.81, P<0.001). The survival rate of men after hip fracture is lower than that of women, the number of people without spouse is lower than that of people with spouse, the number of people with more than 70 years old is lower than that of people with less than 70 years old, and the number of complications ≥2 people is lower than that of people with less than 1 complication, all of which are statistically significant ( P<0.05). Conclusion:The survival rate within 9 months after the occurrence of hip fragility fracture decreased significantly, and it needs to be tracked and managed for at least 9 months to effectively reduce the risk of death; Male, no spouse, age>70 years old, number of complications ≥2 will increase the risk of death after hip fragility fracture, leading to reduced survival rate of patients.