Objective To evaluate the effects of hydrogen-rich saline on chronic inflammatory pain in rats.Methods Thirty-two male Sprague-Dawley rats,aged 8-10 weeks,weighing 200-250 g,were randomly divided into 4 groups (n =8 each) using a random number table:control group (group C);complete Freund's adjuvant (CFA) group;hydrogen-rich saline group (group H2);CFA + hydrogenrich saline group (CFA+H2 group).Chronic inflammatory pain was induced by injecting CFA 100 μl into the plantar surface of the left hindpaw in CFA and CFA + H2 groups.In H2 and CFA + H2 groups,0.6 mmol/L hydrogen-rich saline 5 ml/kg was injected intraperitoneally once a day for 7 consecutive days starting from 1 day after injection of CFA,while the equal volume of normal salinc was given instead of hydrogen-rich saline in C and CFA groups.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before CFA injection and 1,3 and 7 days after CFA injection.The rats were sacrificed after the last measurement of pain threshold on day 7 after CFA injection.The left lumbar segments (L4 5) of the spinal cord were removed for determination of nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) expression by Western blot.Results Compared with C group,no significant change was found in the MWT,TWL and expression of Nrf2 and HO-1 in H2 group,and the MWT was significantly decreased,the TWL was shortened,and the expression of Nrf2 and HO-1 was up-regulated in CFA and CFA+H2 groups.Compared with CFA group,the MWT was significantly increased,the TWL was prolonged,and the expression of Nrf2 and HO-1 was up-regulated in CFA+H2 group.Conclusion Hydrogen-rich saline can alleviate chronic inflammatory pain in rats,and activation of Nrf2/ARE signaling pathway in the spinal cord is involved in the mechanism.

Objective To investigate the effects of hyperbaric Oxygen (HBO)exposure on Caspase-3 and Bcl-2 mRNA ex-pressions in both internal carotid artery (ICA)and basal artery (BA)in rabbits.Methods Twenty-four healthy adult New Zealand rabbits were randomly divided into 2 groups:HBO group and the control group,with each group consisted of 12 animals.The rab-bits in the HBO group were exposed to HBO at 2.2 ATA for 60 minutes each day for 3 successive days.The rabbits in the control group were normally fed without any treatment.Real-time PCR was used to detect Caspase-3 and Bcl-2 mRNA expressions in both ICA and BA in 2 groups.Results HBO significantly decreased the Caspase-3 mRNA expression [(0.038 ±0.006 )vs .(1.000 ± 0.225)]and increased the Bcl-2 mRNA expression [(1.877 ±0.1 69)vs .(1.000 ±0.364)].In ICA,HBO similarly decreased the Caspase-3 mRNA expression [(0.41 9±0.091)vs .(1.000 ±0.1 75)]and increased the Bcl-2 mRNA expression [(1.269 ±0.270) vs .(1.000±0.1 1 7)]in BA.All the differences mentioned above were of statistical significance (P <0.01).Conclusion HBO ex-erts an inhibition effect on apoptosis in cerebral vascular endothelial cells.The mechanism may be related to inhibiting the expres-sion of Caspase-3 mRNA and promoting the expression of Bcl-2 mRNA.

Antithrombin III (AT III) is the most important anti-clotting substance. Recombinant human antithrombin III (rhAT III) expressed in transgenic goat milk attracts more and more attention. Develop an effective purification route for rhAT III is vital to its industrial production. An efficient purification method was developed for the rapid purification of rhAT III by isoelectric precipitation and heparin affinity chromatography. First, casein was effectively removed by isoelectric precipitation. rhAT III was further purified by heparin affinity chromatography. In the process of heparin affinity chromatography, the effects of pH and temperature on the stability of rhAT III were studied, and the effects of operating conditions, elution gradient, flow rate and sample loaded, on the purification efficiency were also studied. Under the optimized conditions, the protein recovery of rhAT III was about 90% with purity over 99%, while its activity recovery was about 50%. Such a purification process is very simple and effective, and it would provide a valuable reference for the further scaling-up of industrial production.
Animals ; Animals, Genetically Modified ; Antithrombin III ; biosynthesis ; Chromatography, Affinity ; Female ; Goats ; Heparin ; Humans ; Mammary Glands, Animal ; metabolism ; Milk ; chemistry ; Recombinant Proteins ; biosynthesis

Objective:To evaluate the application value of electrical impedance tomography(EIT)imaging combining bedside bronchoscopy sputum suction by observing the changes of pulmonary ventilation, tidal volume and dynamic pulmonary compliance after bedside bronchoscopy sputum suction in elderly stroke-associated pneumonia(SAP).Methods:A randomized controlled study was conducted.Patients with SAP admitted to the respiratory intensive care unit of Henan Provincial People's Hospital from January 2017 to December 2018 were enrolled as research objects.They were divided into the control group versus observation group with the only difference in receiving bedside bronchoscope sputum suction replacing control's receiving conventional sputum suction.Impedance imaging region of interest 4(ROI4)values collected by using EIT at admission and 1, 3, 5 days after fiberoptic bronchoscope sputum suction were compared between the two groups.Meanwhile, the tidal volume, dynamic lung compliance, the duration of mechanical ventilation and hospitalization time in intensive care unit were recorded in the two groups.Results:A total of 78 patients meeting an inclusion and exclusion criterion were enrolled, with 37 cases in the control group and 41 cases in the observation group.Compared with control group, the bronchoscope sputum suction group showed the significantly increased regional gas distribution values(2.24±0.77% vs.0.49±0.65%, 7.05±0.77% vs.2.49±0.87%, 12.34±1.47% vs.5.57±0.50%, t=10.85, 24.56 and 26.54, respectively, all P<0.001)at 1, 3, 5 days after fiberoptic bronchoscope sputum suction.The tidal volume and dynamic lung compliance were significantly higher in the observation group than in the control group at 1, 3, 5 days after fiberoptic bronchoscope sputum suction.The duration of mechanical ventilation and hospitalization time in the intensive care unit were shorter in the observation group than in the control group(12.22±0.88 d vs.14.65±0.92 d, 18.41±1.12 d vs.21.14±1.06 d, t=11.91 and 11.01, both P< 0.001). Conclusions:For patients with SAP, an intermittent bedside fiberoptic bronchoscope sputum suction can effectively improve the pulmonary ventilation in the dorsal area, optimize pulmonary respiratory dynamics, facilitate the early withdrawal of the mechanic ventilation, and shorten the hospitalization time in the intensive care unit.

Objective: To explore the injury pattern and features of peripheral nerve in congenital muscular dystrophy patients caused by LAMA2 gene mutation. Method: Seventeen patients genetically or molecular pathologically diagnosed as LAMA2-related congenital muscular dystrophy were recruited in Peking University First Hospital between 2002 and 2015. All the patients received nerve conduction velocity (NCV) and needle electromyography tests. Clinical and laboratory examination data of the patients was retrospectively analyzed. The correlation between the NCV and disease course was determined by Pearson correlation analysis. Additionally, one patient underwent a sural nerve biopsy. Result: Among these 17 identified patients (13 male and 4 female), all of them were diagnosed as congenital muscular dystrophy, and all of them underwent electrophysiological examination at ages between 1 month to 6 years. Electromyogram indicated seventeen patients of myogenic damage, of whom 10 cases were complicated with reduced NCV. Twenty-six of 95 analyzed nerves showed NCV slower than the normal average of contemporary in 17%-47%. Correlation analysis between NCV and the disease course indicated that NCV of median nerves, ulnar nerves, tibial nerves and common peroneal nerves were negatively associated with the disease course (r=-0.737, -0.771, -0.540 and -0.682, respectively; all P<0.05). Sural nerve biopsy revealed peripheral neuropathy changes of myelin. Conclusion There is peripheral nerve injury in LAMA2-related muscular dystrophy patients. It mainly manifests as demyelinating lesions. Moreover, the NCV of peripheral nerve will decrease with the increase of the course of the disease.

Objective:To analyze the clinical features and genotypes of mitochondrial complex Ⅰ deficiency due to NDUFAF5 gene mutations.Methods:Clinical data of 2 cases with mitochondrial complex Ⅰ deficiency due to NDUFAF5 gene mutations admitted in the Department of Pediatrics, Peking University First Hospital from February 2015 to July 2018 were retrospectively reviewed and followed up.Reported cases of mitochondrial complex Ⅰ deficiency due to NDUFAF5 gene mutations were searched in online databases, including the PubMed, Wanfang, Chinese Journal Full-Text Database and VIP database from January 1975 to February 2020 with " NDUFAF5" as the key word.Through literature review, clinical features and genotypes of mitochondrial complex Ⅰ deficiency due to NDUFAF5 gene mutations were summarized.Results:Case 1 showed mentor and mental regression after infection at the age of 1 year and 4 months.The condition of case 1 remained stable at the age of 5 year and 6 months at the last follow-up.Brain magnetic resonance imaging (MRI) showed multiple lesions in the white matter of the frontal and parieto-occipital lobes, basal ganglia, thalamus, cerebellum, brain stem and corpus callosum.Case 2 showed rapidly bilateral visual impairment at the age of 7 years and 4 months.The patient′s vision moderately recovered at the age of 8 years and 8 months.Brain MRI showed midbrain, periaqueductal gray, medulla oblongata and putamen lesions.Spinal MRI showed continuous lesions in the cervical cord 1-4.Genetic test showed NDUFAF5 gene c. 764C>T (p.Ala255Val) and c. 508C>T (p.Arg170Trp), homozygous c. 836T>G (p.Met279Arg) mutations in case 1 and case 2 respectively.Through online searching, 6 reports involving 14 cases were retrieved.The most common clinical phenotype was Leigh syndrome.Two cases had disease onset during the neonatal period, and their disease progressed rapidly and died within 1 year old.Eleven cases had onset during the infantile period, and 72.7% (8/11 cases) of them had a normal development.The common initial symptoms were mental or motor regression, feeding difficulty and dystonia.Seventy-two point seven percent (8/11 cases) had acute/subacute onset after infection, showing paroxysmal deterioration, and died in infancy or childhood.One patient developed dystonia in childhood and visual impairment in adulthood.Conclusions:The onset age ranged from neonatal period to childhood in patients with NDUFAF5 gene mutations, and their clinical phenotypes vary a lot.The main clinical phenotype is Leigh syndrome.Disease onset during the infantile period is frequent, and mostly presents paroxysmal deterioration after infection, while disease onset in childhood is rare.

Objective:To systematically analyze the influence of early removal of urinary catheters on urinary complications in middle-aged and elderly patients after transurethral resection of the prostate.Methods:Randomized controlled trials or clinical controlled trials on early removal of urinary catheters in patients after transurethral resection of the prostate were retrieved from PubMed, Embase, the Cochrane Library, the Web of Science, CNKI, Wanfang Data, VIP database and CBM.RevMan 5.3 was used to analyzed the data.Results:Nine randomized controlled trials and one controlled clinical trial involving a total of 1529 patients were included.The results of meta-analysis showed that there was a significant difference between catheter removal within three days after surgery and removal 4-7days after surgery in the incidence of urinary tract infections[ OR=0.34, 95% CI(0.20-0.58), P<0.01], but there was no significant difference in secondary hemorrhage[ OR=0.86, 95% CI(0.44-1.66), P>0.05].There was no significant difference in the incidence of re-catheterization or secondary hemorrhage between ≤24 hours and 2-3 days after surgery[ OR=1.32, 95% CI(0.57-3.06), P>0.05; OR=3.18, 95% CI(0.32-31.56), P>0.05]. Conclusions:Early postoperative catheter removal(within 3 days)has a clear advantage in reducing the incidence of urinary tract infections, and urinary catheter removal within 24 hours does not increase the incidence of re-catheterization or secondary hemorrhage compared with removal after 24 hours.

Objective: To observe the effect of ulinastatin combined with glutamine on early hemodynamics in patients with severe burns. Methods: Thirty-two patients with severe burns who met the inclusion criteria and hospitalized in the Affiliated Huaihai Hospital of Xuzhou Medical University from January 2016 to December 2018 were selected for conducting a prospective randomized controlled trial. According to the random number table, the patients were divided into conventional treatment group (4 males and 4 females), ulinastatin group (5 males and 3 females), glutamine group (5 males and 3 females), and ulinastatin+ glutamine group (4 males and 4 females), with ages of (36±8), (34±8), (35±9), and (38±13) years in turn. From post injury day 2, patients in the 4 groups were given nutritional support of equal nitrogen and equal calories, of which protein was 2.0 g/kg daily. In addition, patients in the ulinastatin group received intravenous injection of 100 kU ulinastatin every 8 hours for 7 consecutive days; 0.3 g/kg of protein given to patients in the glutamine group was provided by alanine glutamine for 7 consecutive days; patients in the ulinastatin+ glutamine group received corresponding treatments of both ulinastatin group and glutamine group. With the help of pulse contour cardiac output (PiCCO) monitoring technology, the cardiac index, stroke volume index (SVI), global end-diastolic volume index (GEDI), systemic vascular resistance index (SVRI), extravascular lung water index (EVLWI), pulmonary vascular permeability index (PVPI) of patients in each group were measured on treatment day (TD) 1, 3, and 7. Data were processed with Fisher′s exact probability method, one-way analysis of variance, analysis of variance for repeated measurement, and Bonferroni method. Results: The cardiac index was low and the SVI value was lower than the normal value on TD 1 in patients of the 4 groups, without statistically significant differences between any two groups (P>0.05), and then they were all gradually increased. On TD 3 and 7, compared with those of the conventional treatment group, the cardiac index and SVI of patients in the other three groups were all increased, and the cardiac index and SVI of patients in the ulinastatin+ glutamine group were significantly increased (P<0.05 or P<0.01). On TD 1, the GEDI of patients in the conventional treatment group, ulinastatin group, glutamine group, and ulinastatin+ glutamine group were at normal low levels, which were (659±58), (661±79), (659±88), and (653±71) mL/m² respectively, without statistically significant differences between any two groups (P>0.05), and then they all gradually increased. On TD 3 and 7, compared with (684±82) and (742±46) mL/m² of the conventional treatment group, the GEDI of patients in the ulinastatin group, glutamine group, and ulinastatin+ glutamine group were all elevated, which were (732±53) and (777±33), (725±58) and (783±49), (813±65) and (849±27) mL/m² respectively, and the GEDI of patients in the ulinastatin+ glutamine group was significantly increased (P<0.05). The SVRI of patients in the four groups were all at high levels on TD 1, without statistically significant differences between any two groups (P>0.05), and then they all gradually decreased. On TD 3 and 7, compared with those of the conventional treatment group, the SVRI of patients in the other three groups were all increased, and the SVRI in the ulinastatin+ glutamine group was significantly increased (P<0.05). On TD 1, the EVLWI of patients in the conventional treatment group, ulinastatin group, glutamine group, and ulinastatin+ glutamine group were all in the normal range, which were (6.6±0.6), (6.3±0.4), (6.5±0.4), and (6.6±0.6) mL/kg respectively, without statistically significant differences between any two groups (P>0.05), and then they all showed the increasing trend. On TD 3 and 7, compared with (7.1±0.9) and (7.9±0.5) mL/kg of the conventional treatment group, the EVLWI of patients in the ulinastatin group, glutamine group, and ulinastatin+ glutamine group were all decreased, which were (6.2±0.6) and (7.1±0.4), (6.3±1.0) and (7.2±0.9), (5.8±0.7) and (6.7±0.6) mL/kg respectively, and the EVLWI of patients in the ulinastatin+ glutamine group was significantly decreased (P<0.05). On TD 1, the PVPI of patients in the four groups were all in the normal range, without statistically significant differences between any two groups (P>0.05), and then they all gradually decreased. On TD 3 and 7, compared with those of the conventional treatment group, the PVPI of patients in the other three groups were all decreased, and the PVPI in the ulinastatin+ glutamine group was significantly decreased (P<0.05). Conclusions Ulinastatin combined with glutamine can increase the cardiac index, SVI, GEDI, and SVRI and reduce the EVLWI and PVPI in treating patients with severe burns, thereby increasing early cardiac output after injury, promoting tissue and organ perfusion, and reducing pulmonary edema, resulting in significant improvement in early hemodynamics of patients with severe burns.

Objective To evaluate the predicting value of circulating miRNAs for sepsis secondary to pneumonia in elderly patients.Methods From April 2016 to January 2017,44 cases with sepsis secondary to pneumonia,52 elderly patients with pneumonia and 21 healthy older adults as control were involved in this study.The expression levels of MiRNA-150 5p,miRNA-25-3p,miRNA-122 5p and miRNA-223-3p in plasma were evaluated by fluorescence quantitative PCR.The demographic characteristics,sequential organ failure assessment (SOFA)scores,prognosis and days stayed in ICU were recorded.The area under the receiver operating charaeteristic(ROC)curve was used to calculated the specificity and sensitivity of miRNA in identifying sepsis-associated pneumonia.Results There were significantly differences among levels of circulating miRNA-223-3p in pneumonia,sepsis and healthy control groups(F =36.441,P =0.000),△CT values were 2.39 ± 1.36,1.44± 1.43,and 4.58 ± 0.91,respectively.The relative expression levels of miRNA-223-3p in the three groups were significantly different (P =0.000),which were 0.189 (0.107,0.367),0.361 (0.221,0.735),and 0.044 (0.022,0.061),respectively.The AUC of miRNA-223-3p for predicting sepsis from pneumonia was 0.964(95 ％CI =0.925 1.000).At a cutoff value of 2.759,miRNA-223-3p yielded a sensitivity of 82.9％ and a specificity of 100.0％.Conclusions MiRNA-223-3p expression is up-regulated in patients with sepsis secondary to pneumonia compared to that of patients with pneumonia,and it could be used to predict sepsis associated pneumonia.

Objective:To explore the predictive value of HACOR score [heart rate (H), acidosis (A), consciousness (C), oxygenation (O), and respiratory rate (R)] on the clinical outcome of non-invasive positive pressure ventilation in patients with pulmonary encephalopathy due to chronic obstructive pulmonary disease (COPD).Methods:A prospective study was conducted. The patients with COPD combined with pulmonary encephalopathy who were admitted to Henan Provincial People's Hospital from January 1, 2017 to June 1, 2021 and initially received non-invasive positive pressure ventilation were enrolled. Besides non-invasive positive pressure ventilation, standard medical treatments were delivered to these patients according to guidelines. The need for endotracheal intubation was judged as failure of non-invasive ventilation treatment. Early failure was defined as the need for endotracheal intubation within 48 hours of treatment, and late failure was defined as the need for endotracheal intubation 48 hours and later. The HACOR score at different time points after non-invasive ventilation, the length of intensive care unit (ICU) stay, the total length of hospital stay, and the clinical outcome were recorded. The above indexes of patients with non-invasive ventilation were compared between successful and failed groups. The receiver operator characteristic curve (ROC curve) was drawn to evaluate the predictive effect of HACOR score on the failure of non-invasive positive pressure ventilation in the treatment of COPD with pulmonary encephalopathy.Results:A total of 630 patients were evaluated, and 51 patients were enrolled, including 42 males (82.35%) and 9 females (17.65%), with a median age of 70.0 (62.0, 78.0) years old. Among the 51 patients, 36 patients (70.59%) were successfully treated with non-invasive ventilation and discharged from the hospital eventually, and 15 patients (29.41%) failed and switched to invasive ventilation, of which 10 patients (19.61%) were defined early failure, 5 patients (9.80%) were late failure. The length of ICU and the total length of hospital stay of the non-invasive ventilation successful group were significantly longer than those of the non-invasive ventilation failure group [length of ICU stay (days): 13.0 (10.0, 16.0) vs. 5.0 (3.0, 8.0), total length of hospital stay (days): 23.0 (12.0, 28.0) vs. 12.0 (9.0, 15.0), both P < 0.01]. The HACOR score of patients at 1-2 hours in the non-invasive ventilation failure group was significantly higher than that in the successful group [10.47 (6.00, 16.00) vs. 6.00 (3.25, 8.00), P < 0.05]. However, there was no significant difference in HACOR score before non-invasive ventilation and at 3-6 hours between the two groups. The ROC curve showed that the area under the ROC curve (AUC) of 1-2 hour HACOR score after non-invasive ventilation for predicting non-invasive ventilation failure in COPD patients with pulmonary encephalopathy was 0.686, and the 95% confidence interval (95% CI) was 0.504-0.868. When the best cut-off value was 10.50, the sensitivity was 60.03%, the specificity was 86.10%, positive predictive value was 91.23%, and negative predictive value was 47.21%. Conclusions:Non-invasive positive pressure ventilation could prevent 70.59% of COPD patients with pulmonary encephalopathy from intubation. HACOR score was valuable to predict non-invasive positive pressure ventilation failure in pulmonary encephalopathy patients due to COPD.