Through the doctor group, the questionnaire survey to understand what the doctor online habits situ-ation of network communication, doctors and patients, the effect of the real network environment for medical diag-nosis and treatment, news media under the network environment on the influence of the doctor and the doctor group about web development comprehensive view of the impact of the doctor-patient relationship.Points out that the current network environment brings to the doctor-patient relationship: the influence of the Internet become some doctors information update and the accumulated knowledge source, network has become an important platform inter-action some doctors and patients, doctors need to deal with an increasing number of network patients, the doctor-shave reactive shifts to the media negative reports.

Objeetive To analyze the causes and to explore prevention and management of bile leakage after laparoscopic common bile duct exploration with choledochoscopy followed by primary suturing of choledochal incision.Methods The clinical data of 52 patients with bile leakage after laparoscopic common bile duct exploration choledochoscopy and primary suturing of choledochal incision carried out for choledocholithiasis between June 2011 to June 2013 were retrospectively studied.Results All the 52 patients successfully underwent the laparoscopic surgery and left hospital.The operation time was (101 ± 26) minutes (range 55～ 145 minutes).The intraoperative blood loss was (36±28) ml (range 10～ 100 ml).All the patients were ambulatory after the first postoperative day.The recovery time of postoperative gastrointestinal function was (49.8 ± 12.5) hours (range 37 ～ 74 h).The total hospitalization time was (10.8 ± 2.5) days (range 7 ～ 15 days).The average hospitalization days after surgery was (5.7 ± 1.7) days.The average hospitalization cost was (24 827 ± 3 776) yuan.There were two patients who developed intraoperative bile leakage which was treated with further suturing.Five patients developed postoperative bile leakage and they were cured after unobstructed drainage for 5 days through conservative treatment.After a follow-up of 1 ～ 2years,there was no recurrent lithiasis.The stone clearance rate was 100％.There was no bile duct stricture or other complications.Conclusion In expert hands and with proper selection of patients,laparoscopic common bile duct exploration,choledochoscopy and primary suturing of choledochal incision were safe,effective and feasible for choledocholithiasis.

Objective To study the characteristics of Th17 cells frequency and the relationship with clinical characters in paediatric patients with chronic hepatitis B.Methods Forty-five paediatric patients with chronic hepatitis B within 16-year old from December 2010 to February 2014 in Maternal and Child Health-Care Hospital were enrolled in present study,and 11 age-matched healthy children were enrolled as control group.Hepatitis B virus markers(HBV-M),HBV-DNA loads and alanine amino shift enzyme(ALT) levels in serum were detected in the two groups.The frequencies of Thl7 cells in peripheral blood were measured by flow cytometry.Tumor grade was classed in Children with liver tissue.Results Compared with the control group ((1.61 ± 0.56) ％),the Th17 cells frequencies in peripheral blood ((3.03 ± 1.08) ％) were significantly increased in chronic hepatitis B paediatric patients(t=-0.52,P＜0.01).There was no correlations between Th17 cells frequencies with HBV DNA quantity(lg(HBV-DNA)6.49±3.25) and ALT level((40.5±11.3) U/L) (P ＞0.05).The patients with higher HAI scores (G2-G3) had a greater proportion of Th17 cells ((5.95 ±2.05)％) than those with lower HAI scores(G0-G1) ((1.31±0.19)％),and the difference was significant(t =-1.51,P＜0.01).Conclusion Th17 cells frequencies in peripheral blood were associated closely with liver injury in paediatric patients with CHB.

Objective To study the specificity, sensitivity of diagnostic antigen for Trichinella spiralis(T. s). Methods T668 recombinant protein from highly efficient expression of newborn larvae stage-specific gene of T. s in E. coli as diagnostic antigen. By using negative and positive sera from rabbit, pig, human as the first antibody, and goat-anti-rabbit IgG, goat-anti-pig IgG, goat-anti-human IgG labeled with HRP as the secondary antibody, indirect ELISA method was established for detecting anti-T.s antibody, while excretory-secretory (ES) antigen of T.s muscle larvae was used as control. Results Sera from rabbit, pig and human were detected by T668 recombinant protein as antigen, which showed a positive rate of 100% with 0.016 ?g/well. There was no difference on the results between the T668 recombinant antigen and the ES antigen for diagnosing T. s-infection. Conclusion The T668 recombinant antigen is promising in substituting ES antigen in the detection of anti-T. s antibody.

ObjectiveTo observe the effect of acetylcholine (ACh) on lipopolysaccharide (LPS) induced inflammatory model of rat alveolar macrophages, and to observe the effect of the acetylcholinesterase inhibitor physostigmine (Phy) on the anti-inflammatory effect of ACh.Methods The rat alveolar macrophages NR8383 were cultured in vitro, which were divided into five groups: blank control group, LPS group (stimulated with 1 mg/L LPS for 12 hours), LPS+ ACh group (0.01, 0.1, 1, 10, 100μmol/L of ACh were added for 5 minutes before LPS stimulation), LPS+ Phy group (1 mmol/L Phy was added for 5 minutes before LPS stimulation), and LPS+ ACh+ Phy group (1 mmol/L Phy and 10μmol/L ACh were added for 5 minutes before LPS stimulation). The supernatants were collected in each group, the enzyme-linked immunosorbent assay (ELISA) was used to assay the contents of tumor necrosis factor-α (TNF-α), interleukins (IL-1β, and IL-6). The activity of acetylcholine esterase (AChE ) in the supernatant was also determined.Results① The contents of TNF-α (ng/L: 605.09±57.13 vs. 34.07±8.62), IL-1β (ng/L: 377.09±28.55 vs. 32.33±10.62) and IL-6 (ng/L: 558.04±77.45 vs. 42.62±11.21) in the LPS group were significantly higher than those in the blank control group (allP< 0.05). These results indicated that the inflammatory model of rat alveolar macrophages was constructed successfully.② ACh with the final concentrations of 0.01, 0.1, and 1μmol/L had less influence on the production of TNF-α, IL-1β and IL-6 in the culture supernatants of alveolar macrophages stimulated with LPS compared with LPS group (allP> 0.05). Nevertheless, 10μmol/L and 100μmol/L ACh notably reduced the production of TNF-α (ng/L: 451.19±30.67, 332.19±32.19 vs. 604.96±22.56), IL-1β(ng/L: 261.08±24.78, 143.98±28.39 vs. 367.06±10.44) and IL-6 (ng/L: 342.75±54.60, 235.48±29.75 vs. 562.69±63.34) in the culture supernatants compared with the LPS group (allP< 0.05).③ The activity of AChE in the LPS group was significantly higher than that in the blank control group (kU/L: 5.21±0.63 vs. 3.09±0.10,P< 0.05). The activity of AChE was successfully inhibited by 1 mmol/L acetylcholinesterase inhibitor Phy pretreatment compared with that in the LPS group (1.51±0.12 vs. 5.21±0.63,P< 0.05).④ The level of TNF-α (ng/L: 183.17±35.44 vs. 451.19±30.67), IL-1β (ng/L: 91.49±12.27 vs. 261.08±24.78) and IL-6 (ng/L: 108.17±22.82 vs. 342.75±54.60) in the culture supernatants of LPS+ ACh+ Phy group was significantly decreased as compared with LPS+ ACh group (allP< 0.05).Conclusions ACh with the final concentrations of 10μmol/L and 100μmol/L can inhibit the LPS induced inflammatory reaction in alveolar macrophages. The acetylcholinesterase inhibitor Phy can reinforce the ACh-mediated anti-inflammatory effect on alveolar macrophages inflammatory model.

Objective To investigate the real-time regulatory effects of IFN-γ, programed death ligand 2(PDL2) and janus kinase/signal transducers and activators of transcription (JAK/STAT) signaling pathway on the adherence, proliferation and migration of human placenta-derived mesenchymal stem cells(hPMSCs) based on a finding that IFN-γ could enhance the expression of PDL2 in hPMSCs through JAK/STAT signaling pathway.Methods hPMSCs were isolated by using enzyme digestion method and then co-cultured with IFN-γ, anti-PDL2 monoclonal antibody (anti-PDL2 McAb) and JAK inhibitor, respectively.Real-time cell analysis (RTCA) was used to detect the dynamic changes in the adherence, proliferation and migration of hPMSCs following various interventions.Results IFN-γ remarkably suppressed hPMSCs proliferation during the period from 40 hours to 80 hours after intervention and also inhibited the non-targeted migration of hPMSCs.However, hPMSCs adherence was not affected by IFN-γ.Co-culturing hPMSCs with anti-PDL2 McAb significantly enhanced hPMSCs adhesion and inhibited their non-targeted migration, but had no significant effect on hPMSCs proliferation.Furthermore, the proliferation of hPMSCs co-cultured with IFN-γ and anti-PDL2 McAb was significantly inhibited as compared with that of anti-PDL2McAb treatment group.The adhesion, migration and proliferation of hPMSCs were significantly inhibited after co-culturing them with JAK inhibitor.Conclusion IFN-γ can remarkably suppress the proliferation and migration of hPMSCs.PDL2 can enhance the migration and inhibit the adhesion of hPMSCs.JAK/STAT signaling pathway is involved in regulating the adhesion, migration and proliferation of hPMSCs.

AIM:To investigate the role of microRNA-132 (miR-132) on alveolar macrophage inflammation. METHODS: Rat alveolar macrophage cell line NR8383 was transfected with miR-132 mimic, mimic negative control ( NC) , miR-132 inhibitor, or inhibitor NC.The cells were divided into transfection group, transfection +lipopolysaccha-ride ( LPS) group, and transfection +LPS +acetylcholine ( ACh) group.The mRNA expression of acetylcholinesterase ( AChE) was detected by real-time PCR.The protein levels of AChE, signal transducer and activator of transcription 3 (STAT3) and phosphorylated STAT3 (p-STAT3) in the cells, and nuclear factor-κB (NF-κB) in the cytoplasm and nu-cleus were analyzed by Western blot.The activity of AChE in the culture supernatant was measured by AChE activity assay kit.The nuclear translocation of NF-κB was detected by immunofluorescence assay.RESULTS: Up-regulation or down-regulation of miR-132 had no effect on the mRNA expression of AChE.However, up-regulation of miR-132 decreased the protein level of AChE compared with mimic NC group (P<0.05).Transfection with miR-132 inhibitor increased the pro-tein expression of AChE compared with inhibitor NC group ( P<0.05 ) .In the alveolar macrophages treated with LPS+ACh, the inhibition of nuclear translocation of NF-κB p65 in miR-132 mimic group was more effective than that in mimic NC group ( P<0.05) .The inhibitory effect in miR-132 inhibitor group was weaker than that in inhibitor NC group ( P<0.05 ) .The inhibitory effect of miR-132 mimic on the protein levels of STAT3 and p-STAT3 was stronger than that of mimic NC (P<0.05).CONCLUSION:miR-132 in LPS-stimulated alveolar macrophages reinforced ACh-mediated anti-inflam-matory reaction by targeting AChE to suppress ACh hydrolyzation, which was related to the suppression of NF-κB and STAT3 activation.

Objective To evaluate the risk factors for benign childhood epilepsy with centrotemporal spikes (BECT) complicated by electrical status epilepticus in sleep(ESES).Methods From January 2013 to January 2017,a total of 80 children diagnosed as BECT in pediatric neurology outpatient department of Provincial Clinical Medical College Affiliated to Fujian Medical University were enrolled.According to whether there was an attack of ESES or not,patients were divided into ESES group(38 cases) and non-ESES group(42 cases).In order to elucidate risk factors for BECT complicated by ESES,clinical data including age,gender,first seizure age,seizure frequency before treatment,types of seizure,therapeutic drug,recurrence of seizure after treatment,febrile seizure,status at birth,family history,brain MRI,discharge quantity,discharge location,and intelligence score were investigated by multivariate Logistic regression analysis.Results Compared with non-ESES patients,ESES patients were more likely to have recurrence of epilepsy after treatment (68.4％ vs 26.2％,P ＜ 0.001),and had worse intellectual development (＜ 90 scores;73.7％ vs 38.1％,P =0.001);while electroencephalogram showed more discharge in anterior location (47.4％ vs 21.4％,P =0.014) and bilateral distribution of brain (52.6％ vs 26.2％,P =0.015).However,the multivariate Logistic regression analysis showed that only recurrence of seizure after treatment was the risk factor for ESES in BECT patients(P=0.008,OR=4.039,95％CI:1.429-11.418).Conclusion Recurrence of seizure after treatment of BECT was a high risk factor for ESES.Controlling seizure and reducing ESES phenomenon could be beneficial to alleviate the intellectual impairment of patients with BECT.

Objective:To investigate the expression levels of serum calcitonin (CT) and osteoponin (OPN) in peripheral blood of patients with atrial fibrillation (AF) and their relationship with AF.Methods:A case control study was conducted, 160 AF patients treated in Shaanxi Provincial People's Hospital from June 2020 to December 2021 were selected as case group, and 160 healthy people in the same period were selected as control group. The expression levels of serum CT and OPN in the two groups were analyzed retrospectively, and the correlation between them and AF was analyzed. The value of CT and OPN levels in predicting the occurrence of AF was analyzed by receiver operating characteristic (ROC). Kappa consistency test was used to analyze the efficacy of the combination of them in predicting the occurrence of AF.Results:The serum calcitonin level ((13.5±3.2) ng/L) in the case group was lower than that in the control group ((17.3±3.1) ng/L), and the osteopontin level ((53.8±8.2) μg/L) was higher than that of the control group ((44.1±6.8) μg/L), the difference between the two groups was statistically significant ( t=10.79, P<0.001; t=11.50, P<0.001). The results of binary logistic regression analysis showed that serum calcitonin was the protective factor of atrial fibrillation ( OR=0.723, 95% CI: 0.661-0.790, P<0.001), and osteopontin was the risk factor ( OR=1.183, 95% CI: 1.131-1.237, P<0.001). ROC analysis showed that the area under curve (AUC) of CT, OPN and their combination in predicting AF were 0.794, 0.824, and 0.892, respectively. The predictive critical value for serum CT was <15.0 ng/L and >48.5 μg/L for OPN. The sensitivity, specificity of the combination in AF prediction were 67.50% and 93.75% respectively, and Kappa value was 0.613. Conclusion:The expression of serum CT and OPN was abnormal in patients with atrial fibrillation. The prediction of AF by combined examination of the two had a high degree of consistency with the actual results, which could provide reference for early diagnosis and treatment of AF. However, the rate of missed diagnosis was relatively high, which needs attention in clinical application.

ObjectiveTo determine the pathogens and drug resistance in patients with bacterial diarrhea in Lishui City of Zhejiang Province from 2015 to 2019， and provide scientific evidence for prevention and treatment of bacterial diarrhea. MethodFecal specimens were collected from patients with diarrhea in the People’s Hospital of Lishui City from 2015 to 2019. Bacteria were identified by time-of-flight mass spectrometer and serum agglutination reaction. Drug sensitivity in the suspected bacteria was identified by VITEK 2 Compact system. ResultsA total of 2 937 fecal samples were tested from 2015 to 2019， of which 191 were positive for bacteria. The prevalence was 6.65% in male and 6.32% in female. It was highest in the age group 21‒30 years old， followed by the group 51‒60 years old. Summer was the season with the highest prevalence of bacteria. Furthermore，the bacterial species included salmonella （3.98%）， Vibrio parahaemolyticus （1.43%）， aeromonas （0.48%）， shigella （0.37%） and other bacteria （3.66%）. Salmonella had high resistance to cefuroxime and amikacin. Vibrio parahaemolyticus and shigella had high resistance to ampicillin. Aeromonas had high resistance to ampicillin and ampicillin/sulbactam. ConclusionPrevalence of bacteria differs by gender， age and seasons in patients with bacterial diarrhea in Lishui from 2015 to 2019. Salmonella is the principal pathogen in bacterial diarrhea. Additionally， multiple drug resistance is commonly identified. Therefore， it warrants strengthening the pathogenic surveillance on bacteria and drug resistance in bacterial diarrhea.