OBJECTIVETo explore the effects of down-regulated tryptase expression in mast cells on the synthesis and release of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) of vascular endothelial cells.

METHODSTryptase-siRNA (small-interfering RNA) vector was constructed to inhibit tryptase expression in P815 cells. The medium of P815 cells treated by the tryptase-siRNA (RNAi-P815 group) or pure vector (P815 group) was collected and used to culture bEnd.3 cells. The messenger RNAs (mRNAs) of IL-6 and TNF-alpha in bEnd.3 cells and their protein levels in the medium were measured by reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively.

RESULTSIL-6 and TNF-alpha mRNAs in bEnd.3 cells cultured in RNAi-P815-conditioned medium decreased significantly compared to those in P815-conditioned medium. Consistently, IL-6 and TNF-alpha protein levels in the medium of bEnd.3 of RNAi-P815 group were lower than those of P815 group.

CONCLUSIONReduced tryptase expression significantly inhibited the synthesis and release of IL-6 and TNF-alpha in vascular endothelial cells. RNA interference targeting tryptase expression may be a new anti-inflammatory strategy for vascular diseases.

Animals ; Cell Line ; Culture Media, Conditioned ; Down-Regulation ; genetics ; Endothelial Cells ; enzymology ; secretion ; Gene Expression Regulation, Enzymologic ; Interleukin-6 ; genetics ; secretion ; Mice ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transgenes ; Tryptases ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; genetics ; secretion

OBJECTIVETo evaluate the effect of the aluminum hydroxide (Al-OH) adjuvant on the 2009 pandemic influenza A/H1N1 (pH1N1) vaccine.

METHODSIn a multicenter, double-blind, randomized, placebo-controlled trial, participants received two doses of split-virion formulation containing 15 μg hemagglutinin antigen, with or without aluminum hydroxide (Al-OH). We classified the participants into six age categories (>61 years, 41-60 years, 19-40 years, 13-18 years, 8-12 years, and 3-7 years) and obtained four blood samples from each participant on days 0, 21, 35, and 42 following the first dose of immunization. We assessed vaccine immunogenicity by measuring the geometric mean titer (GMT) of hemagglutination inhibiting antibody. We used a two-level model to evaluate the fixed effect of aluminum Al-OH and other factors, accounting for repeated measures.

RESULTSThe predictions of repeated measurement on GMTs of formulations with or without Al-OH, were 80.35 and 112.72, respectively. Al-OH significantly reduced immunogenicity after controlling for time post immunization, age-group and gender.

CONCLUSIONThe Al-OH adjuvant does not increase but actually reduces the immunogenicity of the split-virion pH1N1 vaccine.

Adjuvants, Pharmaceutic ; chemistry ; Adolescent ; Adult ; Aluminum Hydroxide ; chemistry ; Antibodies, Viral ; blood ; Child ; Child, Preschool ; China ; Data Interpretation, Statistical ; Double-Blind Method ; Female ; Hemagglutination Inhibition Tests ; Humans ; Influenza A Virus, H1N1 Subtype ; immunology ; Influenza Vaccines ; adverse effects ; chemistry ; immunology ; Influenza, Human ; epidemiology ; immunology ; prevention & control ; virology ; Male ; Middle Aged ; Models, Statistical ; Pandemics ; Young Adult

Objective To identify the effect of stereotactic body radiation therapy (SBRT) on the survival of patients with recurrent pancreatic cancer after surgery.Methods The data of 104 patients with recurrent pancreatic cancer after surgery who underwent SBRT in the Department of Radiation Oncology of Changhai Hospital,Navy Medical University from February 2012 to December 2016 were retrospectively analyzed.The prescription doses ranged from 35-40 Gy/4-8 f.Survival analysis was performed using the Kaplan-Meier method,and relevant factors affecting patients' survival were screened by the Cox proportional hazards model.Results The median overall survival (OS) and progression free survival (PFS) was 12.5 (11.0-14.0) months and 7.3 (6.0-8.7) months,respectively,while the 1-year rate of OS and PFS was 55.8％ and 22.1％,respectively.Multivariate analysis indicated that tumor stage,biological effect dose (α/β =10,BED10),the decrease of CA19-9 level after treatment,and follow-up chemotherapy were all related factors affecting overall survival;tumor stage,BED10,the degree of pain relief and the decrease of CA19-9 level after treatment were related factors affecting PFS.Conclusions Patients suffering recurrent pancreatic cancer with early tumor stage,normal CA19-9 level and mild pain before treatment could be better treated by SBRT,BED10 ≥60 Gy and follow-up chemotherapy after radiotherapy can prolong the survival of patients.

Objective:To investigate the current management of nosocomial infection at medical institutions of all levels in Changzhou, so as to provide basis for standardizing nosocomial infections control of hospitals within a medical alliance.Methods:An electronic questionnaire was customized for online survey of 91 hospitals affiliated to eight regional medical alliances in Changzhou city in March 2019. The survey covered such aspects as general conditions of the hospital, profile of nosocomial infection control administrators and other staffing, supervision of hospital nosocomial infection programs, and training needs, as well as outstanding problems and suggestions.Frequency number and percentage represent enumeration data, and χ2 test was used to analyze the in-group differences of medical institutions of three levels. Results:Tertiary public hospitals were superior to the secondary and primary hospitals in organizational structure, professional staffing and target monitoring, with the differences of statistical significance( P<0.05). The most urgent training needs of medical institutions at all levels were knowledge in determination and reporting of infectious diseases/nosocomial infection/infection outbreaks; top imperatives and recommendations were development of operation rules for primary medical institutions and standardization of workflows. Conclusions:Staff of primary medical institutions need capacity building in nosocomial infection control; primary hospitals are equipped with incomplete nosocomial infection control information platform; key departments in general lack homogenous management. Tertiary hospitals are encouraged to play leadership in medical alliances in achieving standardized, homogenous and informationized nosocomial infection control within the medical alliances.

Objective To investigate the effects of cholinergic signal on neural stem cell(NSC)differenti-ation in peri-infarction region after ischemic stroke. Methods Mice were randomly assigned into sham + vehicle group,middle cerebral artery occlusion(MCAO)+ vehicle group,MCAO + donepezil group and MCAO + atro-pine group(n = 25). MCAO was induced by thread-occlusion method. Modified neurological severity score (mNSS)was used to evaluate neurological function recovery,and the brain water content was measured by dry-wet weight method. NeuN/5-bromodeoxyuridine(BrdU),CNPase/BrdU,GFAP/BrdU double-labeled cells were tested by immunofluorescence. Results Brain water content of MCAO + vehicle group was significantly higher than that of sham operation group(P < 0.05). Donepezil-treated MCAO mice had lower neurologic deficit scores and brain water content than of MCAO + vehicle group(P < 0.05). On day 14 and day 28 after MCAO,the NeuN/BrdU, CNPase/BrdU and GFAP/BrdU immune-positive cells of MCAO + vehicle group were markedly increased as com-pared with that of sham+vehicle group(P<0.05).Compared with that of MCAO+vehicle group,the number of NeuN/BrdU-positive cells,CNPase/BrdU-positive cells and GFAP/BrdU-positive cells was higher in MCAO+done-pezil group,and the number of NeuN/BrdU-positive cells and CNPase/BrdU-positive cells of MCAO + atropine group was lower(P < 0.05). Conclusions Cholinergic signal could promote NSCs differentiation in peri-infarc-tion region,a lleviate cerebral edema,and improve the brain function restoration after stroke.

Objective To understand the changing trends of non-communicable diseases (NCDs) in Xinjiang Production and Construction Corps from 1998 to 2008.Methods A stratified-cluster random sampling based cross-sectional NCDs survey was carried out in 2008,and using the data of NCDs from the health service surveys in 1998 and 2004,in Xinjiang Production and Construction Corps.The prevalence rate of NCDs was standardization according to age proportion of the population being surveyed in 1998.Results In 1998,2004 and 2008,the prevalence rates of NCDs in Xinjiang Production and Construction Corps were 17.26%,25.61%,24.85% while the Standardized rates of NCDs were 17.26%,23.54% and 20.49% respectively.The prevalence rates of NCDs were statistically significant different in 35-,45-,55- and over 65 age groups in 1998,2004 and 2008 which showed an consecutive upward trend.The prevalence rates of hypertension,diabetes,cerebrovascular disease,coronary heart disease and chronic obstructive pulmonary disease increased significantly from 1998 to 2008.The prevalence rate of hypertensive disease among 25- age group,diabetes among 35- age group,cerebrovascular disease and coronary heart disease among 45- age groups showed an increasing trend.Conclusion Cardiovascular and cerebrovaseular diseases,together with diabetes were the fastest increasing ones over the past 10 years and becoming the major diseases,making the Xinjiang Production and Construction Corps an aging population.NCDs should be prioritized in the health development plan.Targeted health education should be carried out in the whole population,together with other interventions as well as management programs on chronic diseases to reduce the prevalence of NCDs.

Objective: This study is to repair nostril sill deformity in the cleft lip patients by reconstructing the first auxiliary muscle tension line group and to assess the therapeutic outcome. Methods: 437 cleft lip patients with nostril sill deformity underwent the surgery from January 1, 2012 to November 1, 2016.They were treated using the technique of first auxiliary muscle tension line group reconstruction to repair the deformity. Aesthetic correction evaluations were rated by the GAIS. Random digit was used to randomly select 24 patients during the follow-up for three-dimensional measurement and analysis. The preoperative and postoperative symmetry of the nostril sills were evaluated by paired t test. Results: Mucosal ischemia or wound infection occurred in 11 cases, left obvious scar on the nostril floor. Surgical incisions of the other patients were primary healing. After 6 months to 3 years follow up, GAIS questionnaires of 378 patients demonstrated that 84% patients reported great or moderate improvement of nostril sills. Three-dimensional measurements of 24 patients suggested that there were no significant differences of the nostril sill and the nasal ala between the cleft side and the normal side. Conclusions First auxiliary tension line group reconstruction, which is to restore biomechanical balance rather than merely increase muscle volume, is an effective method of nostril sill repair in the cleft lip patients. Post-operatively, the patients achieved a stable and natural nostril sill, a middle columella, as well as a narrowed nasal ala.

OBJECTIVE To explore the effect of total flavonoids of Rhododendra simsii (TFR) on improving cerebral ischemia/reperfusion injury (CIRI) and its relationship with STIM/Orai-regulated operational Ca2+influx (SOCE) pathway. METHODS Oxygen-glucose deprivation/reoxygenation (OGD/R) PC12 cells were used to simulate CIRI in vitro, and the intracellular Ca2+ concentration and apoptosis rate of PC12 cells were detected by laser confocal microscope and flow cytometry, respectively. The regulation of STIM/Orai on SOCE was analyzed by STIM/Orai gene silencing and STIM/Orai gene overexpression. The CIRI model was established by MCAO in SD rats. The activities of inflammatory cyto?kines IL-1, IL-6 and TNF-αin serum were detected by ELISA. The pathological changes of ischemic brain tissue and the infarction of rat brain tissue were detected by HE staining and TTC staining. The protein and mRNA expression levels of STIM1, STIM2, Orai1, caspase-3 and PKB in brain tissue were detected by Western blotting and RT-qPCR, respectively. RESULTS The results of in vitro experiment showed that the fluorescence intensity of Ca2+ and apoptosis rate in PC12 cells treated with TFR were significantly lower than those in OGD/R group, and this trend was enhanced by SOCE antagonist 2-APB. STIM1/STIM2/Orai1 gene silencing significantly reduced apoptosis and Ca2+overload in OGD/R model, while TFR combined with overexpression of STIM1/STIM2/Orai1 aggravated apoptosis and Ca2+overload. In the in vivo experiment, TFR significantly reduced the brain histopathological damage, infarction of brain tissue, the contents of IL-1, IL-6 and TNF-α in the serum in MCAO rats and down-regulated the expression of STIM1, STIM2, Orai1 and caspase-3 protein and mRNA in the brain tissue, and up-regulated the expression of PKB. The above effects were enhanced by the addition of 2-APB. CONCLUSION The above results indicate that TFR may reduce the contents of inflammatory factors and apoptosis, decrease Ca2+ overload and ameliorate brain injury by inhibiting SOCE pathway mediated by STIM and Orai, suggesting that it has a protective effect against subacute CIRI.

OBJECTIVETo observe X-ray features of before and after treatment of cervical imbalance syndrome with osteopathy and traction intervention in youth patients, then to investigate the clinical effect of the treatment of the cervical imbalances syndrome in youth by osteopathy.

METHODSFrom September 2007 to December 2010, one hundred and eighty-seven students of 19 to 22 years (means 21 years) with neck pain as the main symptom in Beijing university of Chinese medicine were selected and divide them into osteopathy group and traction groups randomly. In osteopathy group, there were 94 patients including 40 males and 54 females; in traction group, there were 93 patients including 42 males and 51 females. The treatment of osteopathy group lasted for 3 weeks,three times per week; traction group by traction treatment three weeks,three times a week, too. X-ray before treatment and 3 weeks after treatment were collected.

RESULTSThe osteopathy group:94 cases, before treatment, abnormal curvature in 57 cases, spinous position change in 45 cases and angular displacement in 44 cases, vertebral sliding in 15 cases, Ruth Jackson line intersect proneness change in 70 cases,stretch change in 47 cases; after treatment, abnormal curvature in 35 cases, spinous position change in 24 cases and angular displacement in 18 cases, vertebral sliding in 3 cases, Ruth Jackson line intersect proneness change in 41 cases, extension change in 33 cases; X-Ray measurement results: cervical curvature improved from (7.070 +/- 4.629) degrees before treatment to (7.660 +/- 4.156) degrees after treatment, angular displacement improved from (13.790 +/- 2.590) degrees before treatment to (11.050 +/- 2.560) degrees after treatment; vertebral sliding improved from (3.770 +/- 0.350) mm before treatment to (3.160 +/- 0.485) mm after treatment. The change of angular displacement and vertebral sliding were so significant before and after treatment in this group, there were statistical defferences between before and after the treatment (P < 0.01) in osteopathy group. The traction group: 93 cases, before treatment,abnormal curvature in 60 cases, spinous position change in 39 cases and angular displacement in 39 cases, vertebral sliding in 15 cases, Ruth Jackson line intersect proneness change in 70 cases, stretch in 47 cases; after treatment,abnormal curvature in 50 cases, spinous position change in 29 cases and angular displacement in 17 cases; vertebral sliding in 3 cases, Ruth Jackson line intersect proneness change in 41 cases, stretch in 33 cases; X-Ray measurement results: cervical curvature improved from (5.590 +/- 4.639) degrees before treatment to (5.990 +/- 4.330) degrees after treatment, angular displacement improved from (13.360 +/- 2.064) degrees before treatment to (11.210 +/- 1.872) degrees after treatment; vertebral sliding improved from (3.790 +/- 0.339) mm before treatment to (3.480 +/- 0.332) mm after treatment. The change of angular displacement and vertebral sliding were also so significant in traction group before and after treatment, there were statistical defferences between before and after the treatment (P < 0.01).

CONCLUSION(1) Pathological changes can be called youth cervical imbalance syndrome. (2) Stress points, angular displacement, cervical vertebral slip, curvature straightened, spinous position change are main X-ray performances. (3) Both osteopathy and traction intervention on are efficient in youth neck pain. (4) Youths cervical vertebra imbalance is early but reversible performance.

Cervical Vertebrae ; diagnostic imaging ; Female ; Humans ; Joint Instability ; diagnostic imaging ; therapy ; Male ; Radiography ; Spinal Diseases ; diagnostic imaging ; therapy ; Traction ; X-Rays ; Young Adult

The last essential enzyme in the biosynthetic pathway of trilobatin, phloretin-4'-O glycosyltransferase (P4'-OGT), catalyzes the conversion of trilobatin to phloretin in vitro. However, only a few P4'-OGTs have been found in plants. This study used Malus domestica phloretin-4'-O glycosyltransferase (MdPh-4'-OGT) as a query to identify and clone two UDP-glucuronosyltransferase (UGT) genes, designated UGT74L2 and UGT74L3, from the transcriptome of Andrographis paniculata. According to a phylogenetic tree analysis, UGT74L2 and UGT74L3 belonged to the UGT74 family, which has been linked to several activities in other species. The in vitro enzymatic reaction demonstrated that UGT74L2 could particularly catalyze the formation of trilobatin from phloretin, but UGT74L3 had no effects. By using Ni-NTA affinity chromatography to extract the soluble UGT74L2 recombinant protein, the enzymatic kinetics of the activity was investigated using phloretin as the substrate. The results showed that the optimal temperature and pH for UGT74L2 enzymatic reaction were 40 ℃ and 8.0 (Tris-HCl system), respectively. Three metal ions (Ca²⁺, Mn²⁺ and Co²⁺) showed inhibitory effect on the activity of UGT74L2, while Mg²⁺ could improve the activity of UGT74L2. Other tested metal ions have no significant effect on UGT74L2. The results of enzymatic kinetic parameters that the K_m value was 29.84 μmol·L^-1, the k_cat was 0.02 s^-1, and the k_cat·K_m^-1 was 572.6 mol^-1·s^-1. By homology modeling, molecular docking and mutation experiments, we found that multiple amino acids residues around the substrate binding pocket play quite an important role during catalytic process, In summary, we identified a novel P4'-OGT gene from medicinal plant Andrographis paniculata and provided a new efficient catalyst to synthesize trilobatin. Meanwhile, this study provides a reference for mining new efficient glycosylation modules from plants.