Objective To determine the difference of macrophage RAW264.7 apoptosis induced by Brucella melitensis virulent strain 16M and attenuated strain M5-90 and elucidate the regulatory role of caspases 3,8 and 9.Methods The best multiplicity of infection (MOI) was determined through kinetic analysis of Brucella melitensis strain 16M and M5-90 induced mouse macrophages apoptosis(bacterium ∶ cell =100 ∶ 1,50 ∶ 1,10 ∶1).The infection model was established using the best MOI =50 ∶ 1.The numbers of in vivo bacteria by colony formation units were calculated after macrophages were infected for different times,including 2,4,8,12,24 and 48 h,and the infected cells were collected.The ratios of apoptosis were detected and the regulation of caspases 3,8 and 9 in apoptosis pathway was elucidated by flow cytometry.Results The numbers of 16M in vivo bacteria were 105.4,104.8,105.8,106.5,108.0 and 109.0,respectively and of M5-90 were 106.1,106.2,106.4,106.3,106.1 and 105.0,respectively.The number of in vivo bacteria of 16M was significantly increased than that of M5-90 after infected for 24 h to 48 h.The ratios of apoptosis induced by 16M after infected for 2,4,8,12,24 and 48 h was (2.67 ± 0.09)％,(13.13 ± 0.30)％,(6.56 ± 0.42)％,(6.49 ± 0.28)％,(16.07 ± 0.86)％ and (24.23 ± 1.67)％,respectively,and by M5-90 was (3.62 ± 0.02)％,(32.01 ± 2.59)％,(17.58 ± 0.44)％,(16.09 ± 0.10)％,(62.53 ± 2.70)％ and (85.53 ± 0.15)％,respectively,and by control group was [(1.90 ± 0.20)％,(1.92 ±0.16)％,(1.99 ± 0.03)％,(2.48 ± 0.11)％,(3.56 ± 0.07)％,(5.26 ± 0.33)％].The differences were statistically between groups in same time.The Brucella melitensis vaccine strain M5-90 was more powerful than virulent strain 16M in respect of inducing macrophage apoptosis after infected for 24 to 48 h.Twenty-four hours after infection,the expression of caspases 3,8 and 9 was (1.47 ± 0.05)％,(1.52 ± 0.02)％ and (2.47 ± 0.12)％,respectively,in control group and the expression was (9.70 ± 0.46)％,(6.08 ± 0.56)％ and (35.08 ± 1.64)％,respectively,after infected for 24 h induced by M5-90.The expression of caspases 3,8 and 9 was significantly higher than that control group (P ＜ 0.01).Twenty-four hours after given caspases 3,8 and 9 inhibitor,apoptosis rate in control group was (66.72 ± 1.28)％,in M5-90 group was (22.58 ± 0.55)％,(53.15 ± 1.85)％ and (29.18 ± 0.23)％,respectively,and compared with control group,apoptosis rate of caspases 3,8 and 9 was significantly lower(P ＜ 0.01).Conclusions Apoptosis of macrophage can be induced by Brucella melitensis virulent vaccine strain 16M and attenuated strain M5-90.M5-90 is stronger than that of strain 16M.Caspases 3,8 and 9 can regulate macrophage apoptosis after M5-90 infection.

Two fragments G1 and G2 of the glycoprotein G gene of bovine respiratory syncytial virus(BRSV) were selected for expression in Escherichia coli based on the analysis of glycoprotein G by DNA Star software.Then the two fragments of glycoprotein G were amplified by PCR with synthesized G gene of BRSV as the template.The amplified fragments G1 and G2 are 570bp and 308bp in length,respectively.The PCR products were cloned into pET30a vector and expressed in soluble form in E.coli after induction of cultured E.coli with IPTG.Both of the recombinant proteins G1 and G2 were purified by immobilized Ni ion affinity chromatography under native conditions.Then the purified proteins were analysed by Western blotting.The results showed that the purified recombinant protein G1 retained good antigenicity and specificity.But the purified recombinant protein G2 didn't possess biological activity.Antibodies against BRSV were detected in suspected bovine serum samples in China by using indirect ELISA and Western blotting with the purified recombinant protein G1.The purified recombinant protein G1 might be used as antigen for establishing serological methods for diagnosis of BRSV infection.And the purified recombinant protein G1 might also be used for preparing polyclonal and monoclonal antibodies for research on biological functions of glycoprotein G of BRSV.

Objective:To evaluate the clinical outcome of botulinum A toxin(BTX-A)injection into external sphincter combined with oral baclofen in treatment of detrusor-external sphincter dyssynergia(DESD)after spinal cord injury(SCI). Methods:A total of 38 urodynamic examination-confirmed DESD patients,male 31 and female 7,with an average age of (36.5?17.8)years old,were included in this study.200 U of BTX-A toxin was dissolved in 8 ml of normal saline and the solution was injected at 8 different sites(1 ml per site)of the external sphincter via a 5F flexible cystoscopic needle.On the second day,9 patients(BTX-A+baclofen group)were randomly selected for baclofen oral administration,3/d for 3 months; the other 26 patients were taken as control.Urodynamic examination was repeated in all patients 4 weeks later;the voiding diary and urodynamic outcomes were compared before and after treatment.The adverse and toxic effects were observed in the patients who were followed up for 2-9 months.Results:One month after treatment the voiding and storing functions of bladder were improved to different degrees,with the mean maximum uroflow rate(Qmax),the mean urine volume,the mean maximal cystometric capacity and the bladder compliance increased significantly and the mean postvoid residual urine volume and the mean maximal voiding pressure decreased significantly(all P

OBJECTIVETo investigate the viral pathogens of acute lower respiratory tract infection (ALRTI) in hospitalized children from East Guangdong Province of China and the relationship of the pathogens with age and seasons.

METHODSThe nasopharyngeal aspirates samples obtained from 345 hospitalized children with ALRTI were investigated for respiratory syncytial virus (RSV), human bocavirus (HBoV), human metapneumovirus (hMPV), influenza virus types A and B, rhinovirus, parainfluenza virus types 1 and 3 and adenovirus by PCR.

RESULTSViral pathogens were detected in 178 patients (51.6%). RSV was the most frequent (19.3%). Novel viruses hMPV (3.2%) and HBoV (3.2%) were found. A highest detection rate (61.9%) of virus was found between January to March. The infants aged 1 to 6 months showed a higher detection rate (71.3%) of virus than the other age groups. The detection rate of viral pathogens was 72.6% in children with bronchiolitis, followed by asthmatic bronchitis (70.0%) and bronchial pneumonia (44.6%).

CONCLUSIONSRSV remained the leading viral pathogens in children with ALRTI in East Guangdong of China. Novel viruses HBoV and hMPV were also important pathogens. The detection rate of viral pathogens was associated with seasonal changes and age. Different respiratory infectious diseases had different viral detection rates, with highest detection rate in bronchiolitis cases.

Acute Disease ; Adenoviridae ; isolation & purification ; Child, Hospitalized ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Metapneumovirus ; isolation & purification ; Nasopharynx ; virology ; Orthomyxoviridae ; isolation & purification ; Respiratory Syncytial Virus, Human ; isolation & purification ; Respiratory Tract Infections ; virology ; Rhinovirus ; isolation & purification ; Seasons

Objective With multi?center investigation, to assess the life quality of patients with maintained hemodialysis (MHD) in Liaoning Province and to explore the relationship among the mineral metabolism, the life quality of the patients with MHD, and the repeated hospitalization within the latest three years. Methods 1192 patients with hemodialysis (at least 3 months) from January to March in 2015 at ten blood purification centers in Liaoning Province were selected for the cross?————————sectional survey. The Kidney Health?related Quality of Life (HRQOL) version 1.3 was used to evaluate the MHD patients' life quality. The total length of hospitalization was divided into four groups: 0 days, 3 to 15 days, 16 to 30 days and above 30 days. Results When serum calcium value ranged from 2.1 to 2.5 mmol/L, kidney?disease component summary (KDCS), mental component summary (MCS), physical component summary (PCS) and SF?36+KDCS corresponded to a higher value (P<0.05). When serum phosphorus value ranged from 1.13 to 1.78 mmol/L, KDCS and SF?36+KDCS corresponded to a higher value (P<0.05). When the calcium phosphorus product value ranged from 40.68 to 49.94, MCS corresponded to a higher value (P<0.05). KDCS showed a linear correlation with age (P<0.001), dialysis age, serum calcium (less than or equal to 2.5 mmol/L) (P<0.05); PCS showed a linear correlation with age (P<0.001) and dialysis age (P<0.05); SF?36+KDCS showed a linear correlation with age (P<0.001), and serum calcium (less than or equal to 2.5 mmol/L) (P<0.05), while age and dialysis age were negatively correlated. The hospitalization days showed a linear correlation with age, dialysis age (P<0.001) and serum phosphorus, calcium phosphorus product value (P<0.05), while dialysis age and calcium phosphorus product value were negatively correlated. Among different groups of total hospitalization days in three years, age, hemodialysis age, serum calcium, serum phosphorus, calcium?phosphorus product value and quality of life values were all statistically significant (P<0.05). Conclusions The life quality of patients with MHD were correlated with serum calcium, phosphorus, calcium and phosphorus product value, iPTH, dialysis age and age, while age and dialysis age were of negative correlation. The total number of hospitalization days in 3 years was closely linearly correlated with age and dialysis age, significantly correlated with serum phosphorus, calcium and phosphorus product value, while dialysis age, calcium and phosphorus product value were in a negative correlation. The total number of hospitalization in 3 years was correlated with the patients' age, dialysis age, serum calcium, serum phosphorus, calcium and phosphorus product value and quality of life.

Two new labdane diterpenoids were isolated from 95% ethanol extract of the leaves of Callicarpa formosana Rolfe by using silica gel column, MCI column, ODS column and HPLC. Their structures were elucidated by HR-ESI-MS, NMR and ECD spectral data. All of them are new compounds, named 13E-6β-hydroxylabda-8(17),13-dien-15-oic acid (1) and 13E-7α-hydroxylabda-8(17),13-dien-15-oic acid (2). Compounds 1 and 2 were tested for antioxidant activity, and none of them had obvious activity.

Objective:To mining differential expression genes (DEGs) and establish a regulatory network of dysregulated microRNAs (miRNAs) and messenger RNAs (mRNAs) in biliary atresia (BA) spectrum via bioinforma-tics analysis, and to explore the pathogenesis of BA.Methods:GSE46960 dataset was download from gene expression omnibus (GEO). DEGs between normal liver tissues and BA tissues were analyzed using the GEO2R analysis tool.The functional and pathway enrichment analyses of DEGs were performed utilizing the Database for Annotation, Visualization and Integrated Discovery (DAVID6.8). A protein-protein interaction (PPI) network was constructed using the PPI database (STRING11.0) and Cytoscape_v3.7.1 software, and thus key genes were analyzed.BA-related miRNAs were obtained using the human miRNA disease database (HMDD_V3.0) and target mRNAs were predicted by the miRNA target prediction database (miRDB). The intersection between the predicted target mRNAs and DEGs from the GSE46960 dataset was selected.The regulatory network of miRNA-mRNA was constructed using Cytoscape software.Results:A total of 565 DEGs, including 352 up-regulated ones and 213 down-regulated ones were identified.Among them, up-regulated DEGs were enriched in extracellular matrix(ECM)-receptor interaction, focal adhesion kinase (FAK), Amoebiasis, and the phosphoinositide 3 kinase/protein kinase B(PI3K/Akt) pathway.Down-regulated DEGs were enriched in metabolic signaling, biosynthesis of antibiotics and steroid biosynthesis pathway.From the PPI network, 10 key genes were screened out.A complex miRNA-mRNA regulatory network was constructed based on screened DEGs.Conclusions:Identified DEGs and miRNA-mRNA regulatory network constructed in this study may help clarify the molecular mechanisms of BA.This study provides a new direction to explore promising molecular targets for the diagnosis and treatment of BA.

In the face of the severe challenges posed by chronic illnesses, patients not only experience negative emotions due to their condition but also undergo positive transformations, such as a sense of benefit finding. This article summarizes the theoretical foundations, assessment tools, influencing factors, and intervention measures related to benefit finding among chronic disease patients. The aim is to provide references for healthcare professionals to develop and implement personalized psychological nursing care for chronic disease patients.

Psoriasis and atherosclerosis are both complex and refractory.Genetic susceptibility,inflammatory response and the role of adipokines may be the main mechanisms of their comorbidity.TCM believes that heat,toxicity,phlegm,and blood stasis are the basic pathological factors of the comorbidity,which intertwine and lead to a longer course of disease.TCM,starting from the holistic concept,has a certain efficacy in the treatment of psoriasis and atherosclerosis,and has received widespread attention.This article reviewed the pathogenesis of psoriasis and atherosclerosis comorbidity from the aspects of heredity,inflammatory response,and the role of adipokines,and combed the current situation of the research on the mechanism of the treatment of psoriasis and atherosclerosis comorbidity with TCM monomer and compounds,providing references for related mechanisms and clinical research.

Objective To study the clinical efficacy of two-stage bone transport combined with induced membrane technique in the treatment of post-traumatic tibial osteomyelitis.Methods A retrospective study was conducted of the 14 patients with post-traumatic tibial osteomyelitis who had been treated by two-stage bone transport combined with induced membrane technique from February 2009 to August 2015.They were 12 males and 2 females,aged from 26 to 59 years (average,40.4 years).By the Cierny & Mader classification,they all belonged to type Ⅳ.Their disease course ranged from 6 to 30 months,averaging 15.3 months.The number of their previous operations ranged from 2 to 6 times,averaging 3.3 times.In the first stage of operation,tibial extended external fixation was installed according to preoperative design,followed by early debridement and extensive removal of osteomyelitis lesion.The length of bone defect after resection of osteomyelitis lesion was from 4 to 18 cm (average,8.5 cm).10％ vancomycin-contained polymethyl methacrylate (PMMA) bone cement was molded and placed in the bone defects.In the second stage of operation,the PMMA-induced membrane was opened to remove the PMMA bone cement,the bone marrow was drilled through and the induced membrane was then sutured up.Tibial osteotomy was conducted at an appropriate part.The average interval between the first and second stages was from 4 to 7 weeks (average,4.8 weeks).The clinical efficacy was evaluated according to the Paley criteria at final follow-ups.Complications were recorded.Results All the patients were followed up for 15 to 50 months (17.5 months).No relapse of osteomyelitis occurred.The total treatment time ranged from 11 to 19 months,with an average of 14.7 months.The total time for external fixation was from 10 to 18 months,averaging 13.2 months.The external fixator indexes ranged from 31 to 75 day/cm,with an average of 48.5 day/cm.According to Paley criteria at final follow-ups,the clinical efficacy was excellent in 11 and good in 3 cases.Pin tract infection occurred in 9 patients,mal-alignment after removal of external fixation in 6,and malunion or nonunion at the docking site in 5.Conclusions Two-stage bone transport combined with induced membrane technique can cure thoroughly post-traumatic tibial osteomyelitis.Its operation procedures are simple but effective.