Objective:To investigate the effects of dexmedetomidine on perioperative cardiac adverse events in elderly patients with coronary heart disease.Methods:Sixty elderly patients,who were diagnosed as coronary heart disease and underwent gastric cancer operation,were randomly divided into 2 groups (n=30):the dexmedetomidine group (Dex group) and the control group.In the Dex group,dexmedetomidine was administered intravenously at 0.5 μtg/(kg.h) after a bolus infusion at 0.5 μg/kg for 10 min before anesthesia induction.In the control group,equal volume of normal saline was infused instead of dexmedetomidine.The 2 groups received the same anesthesia treatment.The venous bloods were collected at the preoperative 0 h and postoperative 24 h.The concentrations of cardiac troponin (cTnⅠ),N-terminal pro-brain natriuretic peptide (NT-proBNP) and hypersensitive C-reactive protein (hs-CRP) were determined.The ECG was monitored at the above time and the postoperative incidence of cardiac adverse events was recorded.Results:The levels of cTnⅠ,NT-proBNP and hs-CRP in serum were elevated in the 2 groups after the operation.Compared with the control group,the levels of cTnⅠ,NT-proBNP and hs-CRP were significantly decreased in the Dex group (P＜0.05).Compared with the control group,the incidence ofbradycardia were significantly increased,while the myocardial ischemia and tachycardia were significantly decreased in the Dex group during the operation (P＜0.05);the incidence of silent myocardial ischemia and arrhythmia was significantly reduced at 3 days after operation in the Dex group (P＜0.05).Conclusion:Dexmedetomidine could decrease the incidence of cardiac adverse events in elderly patients with coronary heart disease.

Objective To establish a method focusing on regulation of CNN3 gene in the rat hippocampus and help to explore the role of CNN3 gene played in the brain physiology and pathology.Methods One cDNA sequence and three shRNAs targeting CNN3 gene were designed and synthesized.The recombinant lentivirus-mediated expressing and three short hairpin RNA ( shRNA) vectors targeting CNN3 gene in the rats were constructed with engineering technology.All recombinant vectors were intravenously injected into rats hippocampi guided by stereotaxic apparatus.Western blot was performed to explore the best shRNA and to study the changes of CNN3 gene in the rat hippocampus after transfection with the silence and over-expressed vectors.Results The lentivirus-mediated vector expressing CNN3-OE and three shRNA vectors targeting CNN3 gene were successfully constructed.Within eight weeks after transfection, the vectors of CNN3-OE and three CNN3-shRNAs changed the expression of CNN3 gene in the rat hippocampus, in particular, all the protein levels of calponin-3 encoded by CNN3 gene were significantly down-regulated along with the time, with the highest inhibitory rate of 73.6%in the CNN3-shRNA2 group.Significant up-regulation of calponin-3 protein level by 93.88%, was found only on the 14th day after transfection.Conclusions Lentivirus-mediated vectors of CNN3-OE and CNN3-shRNAs may regulate in vivo the CNN3 gene level in the local brain region of rats via stereotactic injection.The study lays a foundation for disease prevention and treatment in the future.

Objective To describe and analyze the clinical and laboratory findings in a group of children diagnosed with scleroderma at Beijing Children's Hospital in the last 10 years.Methods The clinical charts of children with scleroderma in the Rheumatology Department at Beijing Children's Hospital,between January 2002 and October 2013 were reviewed.All of them fulfilled the classification criteria for juvenile sclerodema,both systemic scleroderma (SSc) and localized scleroderma (LS) types.T test was used for comparison between the two groups.Results Forty-six patients were enrolled and were diagnosed as scleroderma.Seven patients(15％) suffered from SSc and 39 patients(85％) were LS.Mean age-at-onset of LS was (5±4) years old.The male to female ratio was 1.2:1.Mean age-at-onset of SSc was (9±4) years old.All patients were female.The lesions found in LS were linear scleroderma (54％),mixed morphea (36％),generalized morphea (8％),and panclerotic morphea (3％).Twenty-six patients had internal organs involved.Three patients with nerve system involvement was found in en coup de sabre (ECDS).Systemic involvement included lung and gastrointestinal tract primarily.The heart,nerve system,kidney,eye involvement was also found.One girl had SSc combined with renal crisis.Antinuclear antibodies were positive in 77％ of LS patients and 100％ of SSc patients.Rheumatic factor was positive in 6 patients (15％),5 patients had joint involvement.Tests for anti-Scl-70 antibodies were positive in 5 (71％) patients with SSc.The most common drugs used were methotrexate and prednisone.Conclusion In this study,LS is common in children.SSc is more severe than LS.Multi-center and large sample study is needed to know the characteristics of juvenile scleroderma in China.

Objective To observe the effects of dehydrocorydaline (DHC) on proliferation and collagen secretion of cardiac fibroblasts (CFs) cultured by high glucose;To provide experimental evidence for clinical application of Rhizoma Corydalis. Methods CFs cultured in vitro with high glucose were made into models. Collagenase and trypsin were used for the combine digestion of CFs from ratsbornin 24 h. 2-4 generation CFs were cultured by high glucose (25 mmol/L), and then 100, 50, 25 mg/L dethydrocorydaline was added for intervention. Cellular morphology of CFs was observed after 24, 48 h. CFs proliferation was detected by MTT method. Cell cycle was assessed via flow cytometry. The levels of collagen Ⅰ and collagen Ⅲ were determined by ELISA. Results CFs began to grow adherence 3 hours after planting, and CFs cultured by high glucose significantly proliferated 24, 48 h later (P<0.05, P<0.01). The percentage of S+G2+M phase CFs increased significantly after 48 h (P<0.01). The secretion of collagen Ⅰ and collagen Ⅲ also increased significantly (P<0.01). After the intervention of DHC, CFs proliferation was significantly inhibited (P<0.01);the percentage of S+G2+M phase CFs decreased (P<0.01);the secretion of collagen Ⅰ and collagen Ⅲ was reduced (P<0.05, P<0.01). Conclusion DHC can reduce CFs proliferation, decrease collagen secretion of CFs cultured by high glucose, and has potential effects of anti-myocardial fibrosis.

OBJECTIVETo develop monoclonal antibodies against the catalytic subunit of human telomerase hTERT for its expression detection of human tumors.

METHODSA dominant epitope in hTERT (peptide hTERT(9))was automatically synthesized based on Fmoc method, and was used to immunize BALB/c mice. Hybridomas were generated and screened by ELISA for specific monoclonal antibodies, and the characterization of which were performed by Western blotting and immunohistochemical staining.

RESULTSAntigenic peptide hTERT(9) was synthesized and confirmed by MALDI-TOF-MS and HPLC analysis. Three hybridoma cell lines secreting anti-hTERT(9) antibodies designated as H4, G8 and A11 were established after primary screening and consequent three rounds of limited dilution. Both of H4 and G8 were IgM, while A11 was IgG1 in isotyping. The competitive assay showed that the antibodies were hTERT(9) specific, and the affinity of G8 was stronger than that of H4 and A11 assayed by affinity ranking. However, in Western blotting, both of H4 and G8 stained an about 123 000 protein band with HeLa and 293 cell extracts but not with normal 2BS cells. Besides, positive staining presented in the nucleus of HeLa, while 2BS was non-reactive immunohistochemically. The sections from paraffin-embedded blocks of 127 cases of human cancer, 40 of precancerous and 19 of benign tumors were in situ stained by G8 antibody, the results showed that the human cancer tissues were 80.31% (102/127) positive in specific nuclear reaction, on the contrary, only a minority of precancerous lesions present weak positive (17.5%, 7/40), and negative in benign tumors (0/19).

CONCLUSIONSThe monoclonal antibodies developed against synthetic peptide were hTERT-specific and could recognize both the native and the denatured form. Thus their use in immunoblotting or immunohistochemistry for detecting the telomerase hTERT expression of cancer cell and tissues was promising.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Binding, Competitive ; Blotting, Western ; methods ; Catalytic Domain ; DNA-Binding Proteins ; Female ; HeLa Cells ; Humans ; Immunohistochemistry ; methods ; Mice ; Mice, Inbred BALB C ; Neoplasms ; enzymology ; pathology ; Telomerase ; chemical synthesis ; immunology

OBJECTIVETo develop a recombinant single domain antibody against hTERT, human telomerase catalytic subunit.

METHODSA previously prepared His-tagged hTERT fusion protein was used as the antigen, and the variable regions in heavy chain (VH) of immunized mice were RT-PCR amplified and cloned into the pCANTAB 5E, a phagemid vector. By transfection, the display library of mouse VH was developed. The candidate clones were selected by affinity panning, and soluble VH were obtained after expression in E. coli, HB2151. The resultant single VH antibodies were characterized on their binding potentials by western blotting.

RESULTSAn about 350 bp VH fragment was amplified from spleen cells of mice immunized by His-tagged hTERT and expressed by phage displayed as VH library. The size of the library was 8 x 10(4). After three rounds of affinity panning, 4 independent clones were chosen and consequently expressed as soluble single domain antibodies (Mr = 16 000). In Western blot analysis, the single domain antibody from 2 of 4 clones proved to react with the His-tagged hTERT fusion protein (Mr = 167 000) without dependence of His-tags and also detect the native hTERT (Mr = 127 000) extracted from the human HeLa cancer cell line. DNA sequencing showed both of the single domain antibodies were encoded by the heavy chain variable region of the mouse.

CONCLUSIONSThe single domain antibodies developed were hTERT recognizable and hTERT specific, thus providing a basis for application of recombinant single domain antibody in inhibition of telomerase activity and anticancer therapy.

Amino Acid Sequence ; Animals ; Antibodies, Monoclonal ; genetics ; immunology ; Base Sequence ; Cloning, Molecular ; Complementarity Determining Regions ; genetics ; DNA-Binding Proteins ; HeLa Cells ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; Mice ; Molecular Sequence Data ; Sequence Analysis, DNA ; Telomerase ; immunology

Objective: To analyze the clinical data of children with rheumatic diseases complicated with osteonecrosis and summarize the clinical characteristics, so as to guide clinical work. Methods: The clinical data of 59 children with rheumatic diseases complicated with osteonecrosis from January 2010 to July 2018 were collected and analyzed retrospectively. Results: Among 59 children with rheumatic diseases complicated with bone infarction, 25 cases were systemic lupus erythematosus (SLE), 4 cases were mixed connective tissue disease, 6 cases were juvenile dermatomyositis, 1 case was Takayasu arteritis, 1 case was leukocy to clystic vasculitis, 13 cases were systemic onset juvenile idiopathic arthritis (SJIA), 1 case was polyarthritis, and 8 cases were juvenile ankylosing spondylitis. The median time from the onset of rheumatic diseases to osteonecrosis onset was 18 (7.00, 38.75) months. A total of 115 joints were involved in 59 children, the most common of which were bilateral hips and knees. Twenty-five were single joint involvement and 34 were multiple joints involvement. There were 37 cases (63%) with vasculitis, 9 cases (15%) with oralulcer, 5 cases (8%) with Raynaud's phenomenon, 31 cases (53%) with Cushing's face, 18 cases (31%) with kidney involvement, 25 cases (42%) with hypertension, and 12 cases (24%) with spinal compression frac- tures. According to statistics, 10 children with osteonecrosis occurred without glucocorticoid intake. The longest duration of glucocorticoid therapy was 13 years, and the average duration was about (27±35) months whensymptomatic osteonecrosis occurred. The median cumulative dose of prednisone was 381.9(209.77, 561.19) mg/kg. Conclusion SLE, SJIA and juvenile ankylosing spondylitis are the three most common rheumatic diseases in children with osteonecrosis. The locations of osteonecrosis are mostly the bilateral hips and knees. It is necessary to strengthen joint examination, physical examination and imaging screening for children with rheumatic diseases after 18 months of onset, so early detection, early treatment are the strategy to improve the prognosis of the diseases.

Objective:To explore the application effect of dyadic intervention scheme based on dyadic disease management theory and Information, Knowledge, Attitude, and Practice model in the discharge preparation of elderly stroke patients and family caregivers.Methods:The 92 pairs of elderly stroke patients and their caregivers hospitalized in the Department of Neurology in People′s Hospital of Zhengzhou University were conveniently selected. The non synchronous control method quasi experimental research was adopted. Totally 46 pairs of subjects who met the criteria for admission and discharge from May to July 2022 were set as the control group, and routine nursing was carried out; from August to October 2022, 46 pairs of subjects who met the criteria for admission and emission were set as the observation group to implement the dyadic intervention program. The scores of discharge readiness, self-efficacy and unplanned readmission rate of patients between the two groups were compared, and the scores of caregiver readiness, self-efficacy and caregiver stress between the two groups were compared.Results:Finally, 85 pairs of subjects completed the study, with 42 pairs in the control group and 43 pairs in the observation group. On discharge day, the total scores of discharge readiness and caregiver readiness in the observation group were (95.19 ± 4.47), (23.02 ± 2.20) points, respectively, which were higher than those in the control group (85.71 ± 5.31), (19.57 ± 1.65) points, with statistically significant differences ( t=8.91,8.16, both P<0.01); the self-efficacy levels of patients in the observation group at discharge and one month after discharge, as well as those of caregivers at discharge and one month after discharge were (73.86 ± 4.87), (75.91 ± 4.51), (75.67 ± 4.99), (79.21 ± 4.90) points, respectively, higher than those in the control group (71.62 ± 5.19), (73.33 ± 4.91), (73.48 ± 4.24), (75.48 ± 4.24) points, with statistically significant differences ( t values were from 2.05 to 3.75, all P<0.05); the pressure levels of caregivers in the observation group at discharge and one month after discharge were (7.51 ± 2.48), (6.28 ± 1.99) points, respectively, lower than those in the control group (8.76 ± 2.55), (7.45 ± 2.36) points, with statistically significant differences ( t=-2.29, -2.48, both P<0.05); the unplanned readmission rate of patients in the observation group one month after discharge was 7.0% (3/43), lower than the control group′s 23.8% (10/42), with statistically significant difference ( χ2=4.65, P<0.05). Conclusions:The implementation of dyadic intervention on elderly stroke patients and caregivers can make their discharge preparation process more adequate, thus reducing the caregiver′s care pressure, reducing the unplanned readmission rate of patients, and improving their health outcomes.

Objective To investigate the risk factors for occult pneumonia(OP) in children with primary nephrotic syndrome(PNS).Methods The clinical data of 115 children with PNS and findings of chest CT from July 2010 to June 2016 at the First Affiliated Hospital of Zhengzhou University were retrospectively analyzed.Based on the findings of chest CT,the subjects were divided into 2 groups:OP group and unoccult pneumonia (UOP) group.The comparisons were made between 2 groups,including gender,age,season,course of disease before admitting to hospital,formation of ascites,white blood cells,C-reactive protein,erythrocyte sedimentation rate,total protein (TP),albumin (ALB),total cholesterol,immunoglobulin G (IgG),immunoglobulin E,urine N-acetyl-beta-D-glucosaminidase (NAG) and 24 h urinary protein quantity/body weight.The single factor analysis was performed to analyze above indicators between 2 groups,and the indicators which had statistical significance were analyzed by single factor analysis were analyzed by the multifactor Logistic regression.The receiver operator characteristic (ROC) curve was drawn to evaluate the predicting ability of the indicators for PNS combined with OP.Results Among 1 15 cases,68 (59.1％) PNS patients were complicated with OP.The result of single factor analysis indicated that the risk factors were the formation of ascites,TP,ALB,IgG and NAG (all P ＜0.05).The multifactor Logistic regression showed that ascites,TP and ALB were the risk factors for OP in children with PNS(P =0.003,0.004,0.003).The area under curve (AUC) of ALB was 0.709,and the critical value was 18.55 g/L(P =0.000);the AUC of TP was 0.658,and the critical value was 39.15 g/L(P =0.004).Conclusion The incidence rate of PNS combined with OP was high.With the presence of formation of ascites,TP ＜39.15 g/L and ALB ＜ 18.55 g/L,it may indicate OP for the PNS children which require special consideration clinically and earlier chest CT examination.