Objective To express recombinant Burkholderia pseudomallei(B.pseudomallei)type Ⅲ secretion system BipD protein,prepare its polyclonal antibodies and verify their immunological traits.Methods The recombinant pET-28a-BipD plasmid was generated,and the pET-28a-BipD-carried E.coli BL21(DE3)bacteria were induced with isopropyl-β-d-thiogalactoside(IPTG)to express recombinant BipD(rBipD)protein.The rBipD was obtained by affinity chromatography using His Trap column,then mixed with Fredrick's adjuvant to immunize BALB/c mice by intraperitoneal injection in order to obtain anti-rBipD polyclonal antibodies.The immunoreactivity of rBipD was detected by Western blot assay using rabbit anti-melioidosis serum and the serum from melioidosis patients.The immunogenicity of rBipD was evaluated using Western blotting and immunofluorescence staining.Finally,rBipD was used to establish an indirect ELISA to detect serum antibodies of clinical melioidosis patients.Results The recombinant plasmid pET-28a-BipD was successfully constructed and transformed into E.coli BL21(DE3)to induce rBipD expression with IPTG treatment.The obtained rBipD had a relative molecular weight of 36×103 and a purity of 95.4％,and had good immunogenicity and immunoreactivity.It could induce the production of specific antibodies after immunizing mice,and mouse polyclonal antibodies against rBipD were prepared with the titer of 1∶512 000.rBipD of 5.0 μg/mL produced specific immune response with the serum of melioidosis patients,but had no specific reaction with the serum of tuberculosis patients,with statistical difference(P＜0.01).Conclusion rBipD with immunological activity is successfully prepared and purified,and its polyclonal antibodies are also developed,which provide a good tool for clinical immunological diagnosis and study of immune mechanism of B.pseudomallei infection.

Purpose: This study aims to evaluate the efficacy and safety of a new combination treatment of vinorelbine and pyrotinib in human epidermal growth factor receptor 2 (HER2)–positive metastatic breast cancer (MBC) and provide higher level evidence for clinical practice. Materials and Methods: This was a prospective, single-arm, phase 2 trial conducted at three institutions in China. Patients with HER2-positive MBC, who had previously been treated with trastuzumab plus a taxane or trastuzumab plus pertuzumab combined with a chemotherapeutic agent, were enrolled between March 2020 and December 2021. All patients received pyrotinib 400 mg orally once daily plus vinorelbine 25 mg/m2 intravenously or 60-80 mg/m2 orally on day 1 and day 8 of 21-day cycle. The primary endpoint was progression-free survival (PFS), and the secondary endpoints included the objective response rate (ORR), disease control rate (DCR), overall survival, and safety. Results: A total of 39 patients were enrolled. All patients had been pretreated with trastuzumab and 23.1% (n=9) of them had accepted trastuzumab plus pertuzumab. The median follow-up time was 16.3 months (95% confidence interval [CI], 5.3 to 27.2), and the median PFS was 6.4 months (95% CI, 4.0 to 8.8). The ORR was 43.6% (95% CI, 27.8% to 60.4%) and the DCR was 84.6% (95% CI, 69.5% to 94.1%). The median PFS of patients with versus without prior pertuzumab treatment was 4.6 and 8.3 months (p=0.017). The most common grade 3/4 adverse events were diarrhea (28.2%), neutrophil count decreased (15.4%), white blood cell count decreased (7.7%), vomiting (5.1%), and anemia (2.6%). Conclusion Pyrotinib plus vinorelbine showed promising efficacy and tolerable toxicity as second-line treatment in patients with HER2-positive MBC.

Objective:To explore the correlation of serum albumin level at admission with clinical prognoses in patients with acute traumatic brain injury (TBI).Methods:One hundred and fifty-four patients with acute moderate-extreme severe TBI (Glasgow Coma Scale [GCS] scores of 3-12 at admission) in Department of Neurosurgery, General Hospital of Western Theater Command from January 1, 2019 to December 31, 2020 were chosen. The comprehensive clinical data of these patients were collected, including age, gender, GCS scores, serum albumin level (hypoalbuminemia defined as<35 g/L), hemoglobin level, comorbidities, treatment measures, and prognoses 6 months after discharge (poor prognosis defined as Glasgow outcome Scale [GOS] scores of 1-2, and good prognosis defined as GOS scores of 3-5). Univariate and multivariate Logistic regressions were used to identify the independent factors for clinical prognoses of these patients, and differences in poor prognosis rate, length of ICU stay, and total hospital cost were compared between different groups.Results:Among the 154 patients, 43 had poor prognosis and 111 had good prognosis. Serum albumin level at admission ( OR=0.916, 95% CI: 0.843-0.996, P=0.001) and GCS scores at admission ( OR=0.701, 95% CI: 0.594-0.828, P<0.001) were independent factors for prognosis. Patients with hypoalbuminemia ( n=70) displayed significantly higher poor prognosis rate, longer ICU stays, and increased total hospitalization cost compared with those without hypoalbuminemia ( n=84, P<0.05); specifically, in patients with GCS scores of 9-12 at admission ( n=58), those with hypoalbuminemia ( n=27) exhibited significantly higher poor prognosis rate, longer ICU stays, and higher total hospitalization cost than their non-hypoalbuminemia counterparts ( n=31, P<0.05); similarly, in patients with GCS scores of 3-8 at admission ( n=96), those with hypoalbuminemia ( n=74) had significantly higher poor prognosis rate than their non-hypoalbuminemia counterparts ( n=22, P<0.05). In patients with good prognosis, those with hypoalbuminemia ( n=56) showed significantly longer total hospital stays, prolonged ICU stays, and increased total hospitalization cost compared with those without hypoalbuminemia ( n=55, P<0.05). Conclusion:Low serum albumin level at admission is likely to lead to poor prognosis, prolonged ICU stays and increased total hospitalization cost in patients with acute TBI.

Objective: To understand the sedentary behavior level of children and adolescents with intellectual disabilities in Jinan City, and to provide a reference basis for developing health behavior intervention strategies. Methods: By used the method of cluster random sampling,the Children s Leisure Activities Study Survey was used to investigate the sedentary behavior level of 285 children and adolescents with intellectual disabilities aged 6-18 years from 7 special education schools in Jinan City. Results: The sedentary behavior time during the whole week among children and adolescents with intellectual disabilities in Jinan City was 394.46 min/d, including 378.00 min/d on weekdays(Monday to Friday) and 388.80 min/d on weekends (Saturday and Sunday), the difference was statistically significant ( Z =-2.19, P <0.05). 80.4%(229) of children and adolescents with intellectual disabilities had sedentary behavior time of more than 2 h/d. The sedentary behavior time per day during the whole week among children and adolescents with intellectual disabilities was negatively correlated with the amount of time spent in moderate vigorous physical activity among them ( r =-0.16, P <0.05). Conclusion Excessive sedentary behavior has become a growing public health concern among children and adolescents with intellectual disabilities,which warrants targeted healthy behavior intervention.

Objective:To explore the diagnosis and treatment of the multi-segment injury of brachial plexus and provide reference for diagnosis and treatment in clinical practice.Methods:From October, 2012 to January, 2015, 24 patients (21 males and 3 females, aged 7-46, average at 25.06±13.01 years) who suffered multi segments injury of brachial plexus were treated by surgical operations. Time from injury to surgery was 7 days to 7 months, with (2.43±2.15) months in average. The general data, injuries and surgical procedures of the patients were recorded. Muscle strength grading was used to evaluate and analyse the curative effect.Results:Twenty-four cases were followed-up for 3.1-7.2 years, with 4.3 years in average. Of the patients, 58.3% of the injuries were caused by mechanical traction. Combined injuries were counted for 83.3%, of which 62.5% combined with ipsilateral limb fractures, 20.8% (5/24) involved in the root of brachial plexus, 79.2% (19/24) with upper part of the clavicle, 91.6% (22/24) with lower part of the clavicle, 16.7% (4/24) with branches of the brachial plexus and 91.7% (22/24) with injuries of 2 segments, 8.3% (2/24) with injuries of 3 segments. At the last followed-up, 55.0% of the patients achieved better than M 3 in total muscle strength. The excellent and good rate was 70.8% in neurolysis group and 42.9% in multiple segment injury group. Conclusion:The mechanism of multi segments injury of brachial plexus is special, and the actual injury is difficult to be located. For patients with multi segments injury, surgical operation should be carried out as early as possible, and the correct surgical procedure can only be determined after the exploration of all sections of the brachial plexus.

The novel coronavirus disease 2019 (COVID-19) outbreak occurred in December last year and spread quickly in the world, causing great harm to people. The rapid progression of this epidemic makes scholars in various fields conduct research on the transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). So far, the wildly accepted routes are droplets and close contact. Controversially, some researchers believe that other routes include aerosol diffusion, fecal-oral transmission, contacting urine, conjunctival infection, and mother-to-infant transmission may also infect people. In this article, combining the newest research and reports, the authors systematically analyzed the theoretical possibility and real-life probability of the transmission routes of the virus in order to help with the research and clinical judgment of the spread of infectious diseases in the future.

Objective:To evaluate the cost effectiveness of primary prophylaxis (PP) with pegylated recombinant human granulocyte colony stimulating factor (PEG-rhG-CSF), PP with recombinant human granulocyte colony stimulating factor (rhG-CSF) and no prophylaxis in women with early-stage breast cancer in China.Methods:Two phase Markov models were constructed for a hypothetical cohort of patients aged 45 with stage Ⅱ breast cancer. The first phase modelled costs and outcomes of 4 cycles docetaxel combined with cyclophosphamide [TC×4, febrile neutropenia (FN) risk>20%] chemotherapy, which assumptions based on literature reviews, including FN rates [base-case (deterministic sensitivity analysis range), 0.29 (0.24-0.35)] and related events [FN case-fatality, 3.4 (2.7-4.1)]. Second phase modelled the long term survival which was link with the relative dose intensity (RDI) [mortality hazard ratio ( HR) of RDI < 85% vs ≥85%, 1.45 (1.00-2.32)]. Clinical effectiveness, therapeutic costs, and economic utilities were estimated from peer-reviewed publications and expert opinions in case of unavailability of published evidences. Results:Compared to rhG-CSF PP and no prophylaxis, the cost of PEG-rhG-CSF PP increased to 5 208.19 RMB and 5 222.73 RMB, respectively. The quality-adjusted life-years (QALYs) enhanced to 0.066 and 0.297, respectively. Accordingly, the incremental cost effectiveness ratios (ICERs) are 79 146.3 RMB and 17 558.77 RMB per QALY, which were both below the willingness to pay (WTP) threshold of three times GDP per capita (18, 000 RMB) recommended by the WHO. Sensitivity analysis suggested that the more clinically effective the primary prophylaxis with PEG-rhG-CSF is, the more cost-effective primary prophylaxis with PEG-rhG-CSF will be. And the lower the mortality HR of RDI<85% vs ≥85% is, the more cost-effective primary prophylaxis with PEG-rhG-CSF will be. Conclusion:Although the cost of PP PEG-rhG-CSF is higher, considering the additional benefits, the administrating of PP PEG-rhG-CSF is likely to be a cost-effective alternative to PP rhG-CSF and no prophylaxis in patients with early stage breast cancer whose FN risks are more than 20% in China.

Objective:To evaluate the cost effectiveness of primary prophylaxis (PP) with pegylated recombinant human granulocyte colony stimulating factor (PEG-rhG-CSF), PP with recombinant human granulocyte colony stimulating factor (rhG-CSF) and no prophylaxis in women with early-stage breast cancer in China.Methods:Two phase Markov models were constructed for a hypothetical cohort of patients aged 45 with stage Ⅱ breast cancer. The first phase modelled costs and outcomes of 4 cycles docetaxel combined with cyclophosphamide [TC×4, febrile neutropenia (FN) risk>20%] chemotherapy, which assumptions based on literature reviews, including FN rates [base-case (deterministic sensitivity analysis range), 0.29 (0.24-0.35)] and related events [FN case-fatality, 3.4 (2.7-4.1)]. Second phase modelled the long term survival which was link with the relative dose intensity (RDI) [mortality hazard ratio ( HR) of RDI < 85% vs ≥85%, 1.45 (1.00-2.32)]. Clinical effectiveness, therapeutic costs, and economic utilities were estimated from peer-reviewed publications and expert opinions in case of unavailability of published evidences. Results:Compared to rhG-CSF PP and no prophylaxis, the cost of PEG-rhG-CSF PP increased to 5 208.19 RMB and 5 222.73 RMB, respectively. The quality-adjusted life-years (QALYs) enhanced to 0.066 and 0.297, respectively. Accordingly, the incremental cost effectiveness ratios (ICERs) are 79 146.3 RMB and 17 558.77 RMB per QALY, which were both below the willingness to pay (WTP) threshold of three times GDP per capita (18, 000 RMB) recommended by the WHO. Sensitivity analysis suggested that the more clinically effective the primary prophylaxis with PEG-rhG-CSF is, the more cost-effective primary prophylaxis with PEG-rhG-CSF will be. And the lower the mortality HR of RDI<85% vs ≥85% is, the more cost-effective primary prophylaxis with PEG-rhG-CSF will be. Conclusion:Although the cost of PP PEG-rhG-CSF is higher, considering the additional benefits, the administrating of PP PEG-rhG-CSF is likely to be a cost-effective alternative to PP rhG-CSF and no prophylaxis in patients with early stage breast cancer whose FN risks are more than 20% in China.

Objective To evaluate the effect and adverse drug reaction of Boulardii yeast combined with triple and quadruple therapy on eradication helicobacter pylori (H.pylori).Methods240 cases of peptic ulcer patients of H.pylori positive were selected in our hospital from January 2014 to December 2015, according to different treatment were divided into the triple threapy group(n=60), the triple therapy union group (n=60), the quadruple therapy group (n=60) and the quadruple therapy union group (n=60).The triple threapy group were given clarithromycin and amoxicillin and pantoraazole;on the basis of this, the triple therapy union group were given Boulardiiyeast.the quadruple therapy group were given clarithromycin and amoxicillin and pantoraazole and CBS capsule, on the basis of this, the quadruple therapy union group were given Boulardiiyeast.The four groups were treated continuously for 14 days.14C-UBT, H.pylori eradication rate and adverse drug reaction in the four groups were evaluated five weeks after treatment.ResultsCompared with the triple threapy group and the quadruple therapy group, H.pylori eradication rate in the triple therapy union group(91.2%) and the quadruple therapy union group(94.7%) were improved obviously, and the adverse drug reactions (31.6%、29.8%) decreased significantly, the cumulative recurrence rate of H.pylori were significantly decreased, and the differences were statistically significant (P< 0.05).ConclusionBoulardii yeast combined with triple and quadruple therapy can obviously increase the H.pylori eradication rate, reduce the incidence of adverse drug reactions and the risk of recurrence.The reasonable treatment plan should be selected according to the actual situation.

OBJECTIVETo investigate the correlation of serum osteoprotegerin (OPG) with the progression of nonalcoholic fatty liver disease (NAFLD) and the noninvasive prediction and diagnosis of nonalcoholic steatohepatitis (NASH).

METHODSA total of 136 patients with NAFLD were enrolled, and their tissue samples for liver biopsy and serum samples obtained at 1 week after liver biopsy were collected; 83 healthy subjects without the symptoms of fatty liver disease proved by ultrasound examination were enrolled as controls. The physiological indicators including height, body weight, and waist circumference were measured, and body mass index was calculated. The biochemical parameters including alanine aminotransferase (ALT), aspartate aminotransferase (AST), AST/ALT, alkaline phosphatase, gamma-glutamyl transferase, total cholesterol, triglyceride (TG), high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol were measured. Double-antibody sandwich enzyme-linked immunosorbent assay was used to determine the serum level of OPG. The rank sum test, chi-square test, t-test, one-way analysis of variance, Spearman correlation analysis, least significant difference test, and receiver operating characteristic (ROC) curve were applied for statistical analysis of various data.

RESULTSSerum OPG level was correlated with AST and TG (P < 0.05), and was highly correlated with hepatocyte fatty degeneration, ballooning degeneration, intralobular inflammation, portal inflammation, and fibrosis degree (P < 0.01). With the increasing NAFLD activity score (NAS), serum OPG level decreased, and there was a highly negative correlation between them (r = -0.928, P < 0.01). Serum OPG level was significantly lower in NASH patients than non-NASH patients. The area under the ROC curve of serum OPG level was 0.963, and according to the Youden index, its optimal sensitivity and specificity were 96.1% and 97.4%, respectively, at an optimal cut-off value of 242.96 ng/L, which suggested a high diagnostic power.

CONCLUSIONIn NASH patients, serum OPG level decreases significantly. Serum OPG level can be used as an independent predictive factor to evaluate NASH and its severity, as well as a noninvasive diagnostic index for NASH.

Alanine Transaminase ; blood ; Alkaline Phosphatase ; blood ; Aspartate Aminotransferases ; blood ; Biopsy ; Body Mass Index ; Case-Control Studies ; Cholesterol ; blood ; Disease Progression ; Enzyme-Linked Immunosorbent Assay ; Fibrosis ; Humans ; Inflammation ; pathology ; Liver ; pathology ; Non-alcoholic Fatty Liver Disease ; blood ; diagnosis ; Osteoprotegerin ; blood ; ROC Curve ; Triglycerides ; blood ; gamma-Glutamyltransferase ; blood