Objective To explore the variation and clinical significance of growth hormone(GH) and insulin like growth factor-Ⅰ(IGF-Ⅰ) in children with critical disease.Methods One hundred and twenty-two cases were recruited into the study,among which 42 cases were sepsis in paediatric intensive care unit (sepsis group),20 cases had finished the cardiopulmonary bypass surgery without infection (surgery group).Sixty healthy children served as control group.Serum levels of GH and IGF-Ⅰ were deteced with immunochemiluminometric assay.Results The serum levels of GH were (6.71 ± 6.62) ng/ml in sepsis group,(8.86 ±8.06) ng/ml in surgery group,(3.87 ± 3.31) ng/ml in control group.Compared with the control group,the serum levels of GH in sepsis group and surgery group were increased significantly (P ＜ 0.05).There was no significant difference between sepsis group and surgery group.The serum IGF-Ⅰ levels were (63.72 ±54.17) ng/ml in sepsis group,(119.06 ± 102.12) ng/ml in surgery group and(154.22 ± 107.10) ng/ml in control group separately.The serum IGF-Ⅰ level of sepsis group was significantly decreased compared to the other two groups (P ＜ 0.05).There was no significant difference between surgery group and control group (P ＞0.05).In sepsis group,compared with those who died,the GH of the survival showed no significant difference,IGF-Ⅰ was significantly increased.(P ＜ 0.05).Conclusion GH playes a positive effect in patients with critical disease.IGF-Ⅰ decreases greatly in patients with serious infection.It suggests that IGF-Ⅰ could be a sensitive marker for reflecting whether there is serious infection.The sharply decreasing of IGF-Ⅰ may predict a bad outcome.

Objective To identify the sequence of cDNA of human leptin coding region, construct a prokaryotic expression vector, and express human leptin in E.coli. Methods RNA was extracted from human placenta tissue. Leptin gene was amplified from RNA by RT-PCR method. The PCR product was ligated with T vector. The ligation reaction was transformed to E.coli DH5? competent cells. The recombinant plasmid was checked by sequencing and restriction analysis. The human leptin gene was cloned into prokaryotic expression vector PET-28a and tranformed into E.coli BL21. The recombinant strain was constructed and induced by IPTG. The product was analyzed with SDS-PAGE and Western blotting. The expressive product was purified by sodium deoxycholate. Results Analysis indicated that the sequence of human leptin cDNA was the same with the reported sequence. Leptin gene was inserted into prokaryotic expression vector. The fusion protein expressed with high efficiency in recombinant E.coli BL21.The results of SDS-PAGE analysis indicated that the molecular weight of the fusion protein was about 20kD. Conclusion Hunam leptin gene was successfully identified from placenta. The leptin gene prokaryotic expression strain was constructed and a high expression of leptin was achieved in E.coli.

Objective To investigate the influence of tethered cord syndrome (TCS) on the up-per urinary tract and its etiology. Methods Forty patients with TCS diagnosed by spinal MRI were enrolled in this study. There were 21 males and 19 females with mean age of 23 years old. The course of disease ranged from 1 to 40 years. Urinalysis, mid-stream urine culture, serum creatinine(SCr), urinary system ultrasound, IVU, eystography and urodynamic study were carried out on all patients. Results Urinary tract infection was found in 17 patients and increased level of SCr was found in 6 pa-tients (251.64±98.5μmol/L). Of the 29 patients who underwent urinary system ultrasound examina-tion, 12 cases had hydronephroais and dilated upper ureter. Of the 30 patients who underwent IVU, 10(33.3%) had ureterectasia and hydronephrosis, 22 cases had bladder turriform or Christmas tree like deformity with diverticulum and trabeculum. Of the 22 patients accepted cystography, 17 cases had vesieoureteral reflux on 27 sides. Post-void residual (PVR) was evaluated in 35 patients and found increased in 31 cases. Cystometry had been done in 33 patients. The mean value of maximal detrusor pressure (Pdetmax) during filling phase was 41.2±20.9 cm H2O. The detrusor compliance was 22.35±18.8 ml/cm H2O. During voiding phase, detrusor-sphincter dyssynergia(DSD)was observed in 16 patients, detrusor areflexia was observed in 16 patients and detrusor underactivity was observed in 13 patients. Resting urethral pressure profilemetry was measured in 16 patients. Maximal urethral closure pressure (MUCP) was 76.1±33.1 cm H2O. The upper urinary tract deterioration was de-fined as increased SCr, hydronephrosis or vesicoureteral reflux. There were 20 patients diagnosed as upper urinary tract deterioration. The compliance of the upper urinary tract deteriorating group and the no-deteriorating group was 9.4±7.8 vs 19.3±15.8 ml/cm H2O, Pdetmax was 43.1±21.2 vs 24.0±11.9 cm H2O, PVR 189.0±138.0 vs 47.8±36.8 ml, MUCP 86.2±32.4 vs 46.8 5±20.8 cm H2O, incidence of damaged detrusor 100.0% vs 69.2% and DSD 65.0% vs 23.1%, respectively. There were significant differences between the 2 groups(P<0.05). And when comparing the VUR group with no VUR group, the incidence of urinary tract infection was 94.1%(16/17) vs 20.0%(1/ 5) (P=0.003). And when comparing urinary tract infection group with no infection group, the inci-dence of upper urinary tract deterioration was 88.2% (15/17) vs 21.7%(5/23)(P=0.000). Condn-sion Low compliance bladder, high Pdetmax during filling phase, increased PVR, high MUCP, damage of detrusor contractive function and DSD are the risk factors for upper urinary tract deteriora-tion in the TCS patients.

Objective To explore prevention of cyclosporine A (CsA) combined with Cobalt protoporphyrin (CoPP) against murine graft versus-host disease (GVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods C57BL/6 (H-2Kb) mice were used as donors and BALB/c (H-2Kd) mice as recipients,which were randomly divided into 4 groups.The mice in total body irradiation group (TBI group) were lethally irradiated and injected intravenously with PBS; The mice in Allo-HSCT group (BS group) were lethally irradiated and injected intravenously with bone marrow cells and spleen cells; The mice in CsA intervention group (CsA group) were injected with CsA intraperitoneally after allo-HSCT; The mice in CsA combine with CoPP intervention group (combination group) received both CsA and CoPP intraperitoneally after alloHSCT.Recipients were monitored for condition,survival rate and weight.The liver,small intestine and skin in the recipients were gained and pathological changes of GVHD were assessed.The kidney was stained with Masson staining dye to observe the tissue fibrosis.The expression levels of renal HO-1 mRNA in the recipients were detected.Results In contrast to BS and CsA groups,GVHD degree in combination group was mild,with less reduction and quick recovery of weight.On the day 30 after HSCT,survival rate in BS group was 36.8％,and that in combination group and CsA group was 69.6％ and 53.5％ respectively (P＜0.05).In comparison with BS and CsA groups,pathological changes in combination group were mild,cellular edema and degeneration degree of the liver,small intestine and skin were slight,and few necrosis and infiltrated inflammatory cells were observed.Tubulointerstitial fibrosis hardly occurred in combination group,but it occurred in CsA group abundantly.As compared with BS group,the expression levels of HO-1 mRNA was increased in combination group,while decreased in CsA group (P＜0.05).Conclusion CsA combined with CoPP enhanced the protective effect of CsA against GVHD,moreover,CoPP could alleviate the side effects of CsA,which might be related with up-regulation of the expression levels of HO-1.

Objective To explore the effect of risk assessment tracking management in reducing unplanned extubation in patients with tracheal intubation.Methods From January to December 2015,120 patients with tracheal intubation in ICU were selected as the control group with routine nursing care of indwelling endotracheal intubation.From January to November 2016,120 patients with tracheal intubation were selected as the observation group,where the risk assessment and risk management were done on the basis of routine nursing as in the control group.The two groups were compared in terms of tracheal intubation and unplanned extubation related knowledge,the implementation of the risk assessment,the accuracy in risk assessment and the incidence of unplanned extubation.Result The tracheal intubation and unplanned extubation related knowledge in the observation group was lower than that of the control group (P＜0.001),The implementation rate of the risk assessment and the accuracy in risk assessment in the observation group were significantly higher than the control group,and the rate of unplanned extubation in the observation group was significantly lower than in the control group (all P＜0.01).Conclusion The implementation on risk assessment and extubation-preventing nursing quality tracing management can enhance the awareness and knowledge of preventing trachea cannula exodus,improve the quality of nursing and reduce the rate of unplanned extubation.

Objective To investigate the outcome of postoperative hypopituitarism and hormone replacement in patients with pituitary adenoma,and to analyze the potential factors related to postoperative hypopituitarism.Methods A total of 215 postoperative patients with pituitary adenoma were analyzed.Pituitary functions( including gonadal,thyroid,and adrenal axes ) were asessed by strict criteria.Data of surgery history and hormone replacement situation were collected for statistical analysis.Results The prevalence of hypopituitarism was 54.0％,including 36.7％ hypogonadism,32.6％ hypothyroidism,and 28.4％ hypoadrenalism.Replacements of gonadal steroid,glucocorticoid,and thyroxine were carried out in 25.6％,84.3％,and 80.6％ of the cases,respectively.Univariate analysis showed that male sex and large tumor were related to hypopituitarism. Conclusion After pituitary adenomectomy,approximately half of the patients present anterior pituitary dysfunction,while quite a number of them have not been treated appropriately.

To investigate the material basis and mechanism of Liupao tea on preventing COVID-19 by network pharmacology and molecular docking.The active ingredients and targets of Liupao tea were searched through the literature and the TCMSP databases and the network between the two was built by Cytoscape 3.7.1.Then using GenCards platform to predict the disease targets,mapping the common targets between Liupao tea and disease.The common targets were imported into the STRING database for exploring the protein-protein interaction.Core targets were enriched by gene ontology (GO) enrichment analysis and KEGG (kyoto encyclopedia of genes and genomes) pathway enrichment analysis using DAVID database etc..Finally,the screened active components were docked with the receptor protein SARS-CoV-2 3CL hydrolase (Mpro).Six active ingredients of Liupao tea were screened,such as (-)-epigallocatechin gallate (EGCG),(+)-catechin,(-)-catechin gallate,α-spinasterol,pelargonidin chloride and squalene,and 156 targets were identified.Among them,there were 112 common targets and 38 core targets with COVID-19.GO enrichment analysis (P<0.01) involved lipopolysaccharide,cell response to hypoxia,etc..And the KEGG pathway enrichment analysis (P<0.01)was conducted to obtain the HIF-1,IL-17,T cell receptor and other signaling pathways associated with COVID-19.The results of molecular docking showed that the active ingredients of Liupao tea were well bound to the receptor protein Mpro.The active ingredients of Liupao tea may control HIF-1,IL-17,T cell receptors signaling pathways by binding Mpro hydrolase and acting on inflammation and immune related targets such as MAPK1,TNF to prevent COVID-19.The EGCG of Mpro activity was determined ,and the IC50 was 3.4 μmol/L,which confirmed that EGCG was a certain inhibition effect on Mpro.

OBJECTIVETo investigate the effect of the lentiviral vector mediated CXCR4 overexpressed mesenchymal stem cell (MSCs) on graft-versus-host disease (GVHD).

METHODSLentiviral vector containing CXCR4 was constructed. CXCR4 overexpressed MSC by lentiviral vector mediated were assessed. A major histocompatibility complex (MHC)-mismatched mouse model of bone marrow transplantation (BMT) from C57BL/6 donors to BALB/c recipients was constructed. Mice were divided into five groups: total body irradiation (TBI) group, mice received irradiation only; BMT group, mice were transplanted with bone marrow (BM) after TBI; GVHD group, mice were transplanted with BM and splencytes after TBI; CXCR4-MSC group, mice were transplanted with CXCR4-MSC, BM and splencytes after TBI; EGFP-MSC group, mice were transplanted with EGFP-MSC, BM and splencytes after TBI. The survival, body weight and clinical score of GVHD in transplanted mice were monitored. Liver, intestine and skin from mice in each group were obtained for histological examination. Plasma concentrations of inflammation factors such as interleukin (IL)-2, IFN-γ and TNF-α were also determined using a cytometric bead array cytokine kit.

RESULTSAll mice in TBI group died within 14 days, while all of BMT group survived. The mean survival times for GVHD, EGFP-MSC and CXCR4-MSC groups were (17.0 ± 2.3) d, (21.7 ± 4.8) d and (30.1 ± 9.1) d, respectively. Treatment with CXCR4 over-expressing MSCs could decrease the mortality rate. All mice in each group developed clinical signs such as hunched posture, dull fur, diarrhea and weight loss. Meanwhile, histopathological findings in target organs were confirmed the presence of GVHD. While, clinical GVHD scores and histopathological scores in CXCR4-MSC group were significantly lower than that of GVHD group. Moreover, compared with control groups, the plasma IL-2, IFN-γ and TNF-α level in recipients infused with CXCR4-MSC were significantly decreased (P<0.05).

CONCLUSIONThe results revealed that CXCR4- transduced MSCs could effectively control the occurrence of mouse GVHD following allogeneic BM transplantation.

Animals ; Bone Marrow Transplantation ; Cytokines ; Genetic Vectors ; genetics ; Graft vs Host Disease ; Lentivirus ; genetics ; Mesenchymal Stromal Cells ; immunology ; Mice ; Receptors, CXCR4 ; genetics ; immunology ; Transplantation, Homologous

OBJECTIVETo investigate ALK gene rearrangements in lung adenocarcinomas in correlation with clinicopathologic parameters including prognosis.

METHODSFluorescence in situ hybridization (FISH) was used to detect ALK gene rearrangements in 53 cases of lung adenocarcinomas. Mutations in exons 18, 19, 20 and 21 of EGFR were analyzed by Scorpion amplification refractory mutation system (Scorpions ARMS).

RESULTSIn a cohort of 53 lung adenocarcinomas, ALK gene rearrangements were identified in 6 tumors (11.3%), including 4 male and 2 female patients. Five were acinar predominant adenocarcinomas and one was mucinous adenocarcinoma (P=1.000). All tumors with the ALK rearrangements had the wild-type epidermal growth factor receptor (EGFR) gene (P=0.023). The median time of disease-free survival (DFS) in ALK positive patients and negative patients were 14 months (95%CI 8.0-20.0) and 31 months (95%CI 24.9-37.1), respectively and the difference was significant (Log-rank test, P=0.019). ALK gene rearrangements were significantly associated with early recurrence, but not tumor size, pathologic stages, histological differentiation and lymph node metastasis.

CONCLUSIONSALK gene rearrangements are present at a higher frequency in lung adenocarcinomas of the Chinese patients. ALK gene rearrangements are mutually exclusive with EGFR mutations and associated with early tumor recurrence.

Adenocarcinoma, Mucinous ; genetics ; pathology ; surgery ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Acinar Cell ; genetics ; pathology ; surgery ; Disease-Free Survival ; Exons ; Female ; Follow-Up Studies ; Gene Rearrangement ; Humans ; Lung Neoplasms ; genetics ; pathology ; surgery ; Male ; Middle Aged ; Mutation ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Receptor Protein-Tyrosine Kinases ; genetics ; Receptor, Epidermal Growth Factor ; genetics