To construct a Klebsiella oxytoca (K. oxytoca)-pBBR1-lux strain that stably express luciferase (lux) gene, and to investigate the distribution of K. oxytoca in vivo and to evaluate the effect of different methods of sterilization.

The luciferase gene cluster (Lux A/B/C/D/E) of pBAV1k-T5-Lux plasmid was amplified, and the pBBR1 plasmid (pBBR1-lux) containing Lux A/B/C/D/E gene cluster was constructed; while Escherichia coli which expressed fluorescein gene group (E. coli-pBBR1-lux) was obtained. The expression of luciferase in E. coli-pBBR1-lux and K. oxytoca-pBBR1-lux were tested using luminescence microplate spectrophotometer. The pBBR1-lux plasmid was electroconverted to the receptive cells of K. oxytoca; after fluorescence intensity identification and three passages of the strain, K. oxytoca-pBBR1-lux which stably express lux genes were screened. Conjugated circular plasmid product was named pBBR1-lux. The value of Luminesence fluorescence signal of E. coli-pBBR1-lux increased significantly compared with E. coli control strain [15 345 (14 676, 18 654) vs. 63 (60, 82)], with significant difference (t = 21.14, P = 0.035). The value of Luminesence fluorescence signal of K. oxytoca-pBBR1-lux [399 303 (265 245, 617 192)] was significantly higher than that of the control strain K. oxytoca [83 (63.5, 86.75)], with significant difference (t = 7.07, P = 0.014). E. coli-pBBR1-lux and K. oxytoca-pBBR1-lux were both able to detect fluorescence signals on Veritas microplate photometer and small animal imager. According to 1︰10 dilution of 6 gradients, the fluorescence detection results showed that the Luminesence value decreased with the decreasing of colony concentration. After repeated passage pBBR1-lux luciferase could be stably expressed in K. oxytoca. Different physicochemical methods had different bactericidal effect on K. oxytoca-producing bacteria. Ultraviolet ray and 84 disinfectant (10%) were the most effective methods to inactivate K. oxytoca-producing bacteria.

To investigate the correlation between human high mobility group protein B1 (HMGB-1), receptor of advanced glycation endproducts (RAGE), C-reactive protein (CRP) and the severity of pulmonary infection after brain trauma surgery and to evaluate the prognosis of the disease.

Total of 182 patients with pulmonary infection after operation of brain trauma who were treated in Affiliated Peace Hospital, Changzhi Medical College from May 2016 to May 2018, prospectively. According to the CURB-65 scoring system for lung infection (CURB-65), patients were divided into mild group (74 cases), moderate group (60 cases) and severe group (48 cases). According to the clinical outcome of 30 days after operation, the patients were divided into survival group (151 cases) and death group (31 cases). The serum levels of HMGB-1, RAGE and CRP in patients with different infection degrees were compared and their correlation between the severity of pulmonary infection were analyzed, respectively. The risk factors of death were determined by Logistic regression analysis and the predictive value of HMGB-1, RAGE and CRP to the death of patients with pulmonary infection after traumatic brain injury were evaluated by receiver operating characteristic curve (ROC).

The level of HMGB-1 of patients in severe group was (10.22 ± 2.35) μg/L, significantly higher than those of moderate group [(3.89 ± 1.01) μg/L] and mild group [(2.00 ± 0.40) μg/L], with significant differences (t = 18.821, 29.502; both P < 0.001). The level of RAGE of patients in severe group was (9.01 ± 2.05) ng/L, significantly higher than those of in moderate group [(5.89 ± 1.20) ng/L] and mild group [(2.12 ± 0.22) ng/L], with significant differences (t = 9.870, 28.722; both P < 0.001). The level of CRP of patients in severe group was (50.33 ± 10.32) mg/L, significantly higher than those of the moderate group [(32.33 ± 8.52) mg/L] and mild group [(15.20 ± 3.52) mg/L], with significant differences (t = 9.930, 27.010; both P < 0.001). The serum levels of HMGB-1, RAGE and CRP of patients in moderate group were all higher than those of mild group, with significant differences (t = 14.744, 26.504, 15.729; all P < 0.001). The serum levels of HMGB-1, RAGE and CRP were positively correlated with the CURB-65 score of pulmonary infection (r = 0.696, 0.763, 0.851; all P < 0.001). The fatality rate of 182 patients was 17.03% (31/182). The proportion of ventilator application, tracheotomy and hypoproteinemia of death patients were higher than those of survival cases, and the age, levels of HMGB-1, RAGE, CRP and procalcitonin (PCT) were higher than those of survival group, with significant differences (all P < 0.05). Logistic regression analysis showed that age, proportion of ventilator application, tracheotomy, hypoproteinemia, HMGB-1, RAGE, CRP and PCT were all risk factors to death (OR = 2.016, 2.423, 2.252, 1.853, 2.606, 2.199, 1.919, 1.904, all P < 0.05). ROC analysis showed that the sensitivity of serum HMGB-1, RAGE and CRP in predicting the death of patients with pulmonary infection after brain trauma surgery were 78.02%, 82.42% and 85.16%, respectively, the specificities were 56.04%, 69.78% and 71.98%, respectively, while the area under AUC were 0.756, 0.801 and 0.882, respectively.

Serum HMGB-1, RAGE and CRP were positively correlated with the severity of lung infection after traumatic brain surgery, and were risk factors for death, which had a high clinical value for predicting death.

To investigate the dynamic changes of activated protein C (APC), brain natriuretic peptide (BNP), acute physiology and chronic health status scoring system Ⅱ (APACHE Ⅱ) in elderly patients with severe pneumonia, and to analyze their relationship with the prognosis.

Total of 288 elderly patients with severe pneumonia who were treated in Chengdu West District Hospital from June 2016 to June 2018 were selected. They were divided into survival group (168 cases) and death group (120 cases) according to 28-day survival condition. The levels of serum APC and BNP of each group werer detected by chemiluminescence and enzyme-linked immunosorbent assay, respectively; and the status of each group were evaluated by APACHE Ⅱ score. The dynamic changes of APC, BNP and APACHE Ⅱ scores in each group at the first day, the fourth day and the seventh day were compared between the two groups, respectively. The risk factors of senile severe pneumonia were explored by Logistic regression analysis, and the value of the combined three indexes in predicting the prognosis of senile severe pneumonia was analyzed by ROC curve analysis.

Compared with the death group, the levels of BNP [(494.62 ± 34.82) pg/ml, (318.42 ± 27.42) pg/ml and (274.61 ± 20.84) pg/ml] and APACHE Ⅱ score [(24.05 ± 4.82), (18.62 ± 3.71) and (12.13 ± 2.62)] of cases in survival group significantly increased at the first day, the fourth day and the seventh day, with significant differences (all P < 0.001), but the levels of APC [(289.34 ± 18.39) ng/ml, (357.64 ±32.71) ng/ml and (427.25 ± 18.45) ng/ml] decreased, with significant differences (t = 5.512, 35.499, 78.552; all P < 0.001). The BNP level and APACHE Ⅱ score of cases in survival group decreased with the extension of hospitalization time (F = 24.538, P < 0.001; F = 12.945, P < 0.001), while the level of APC increased gradually (F = 23.947, P < 0.001). In the death group, with the extension of hospitalization time, the levels of BNP and APACHE Ⅱ score increased gradually (F = 15.302, P < 0.001; F = 10.389, P < 0.001); while the levels of APC decreased gradually, with significant difference (F = 34.165, P < 0.001). There were significant differences in smoking history [36.90% (62/168) vs. 53.33 (64/120)], chronic respiratory disease history [46.43 (78/168) vs. 65.83 (79/120)], partial oxygen pressure [(83.27 ± 6.92) mmHg vs. (76.82 ± 8.65) mmHg] and mechanical ventilation [35.12 (59/168) vs. 52.50 (63/120)] between cases in survival group and death group (χ² = 7.677, P = 0.006; χ² = 10.630, P = 0.001; t = 9.881, P < 0.001; χ² = 8.661, P = 0.003). Logistic regression analysis showed that mechanical ventilation (OR = 4.627, P < 0.001), APC (OR = 2.637, P = 0.012), BNP (OR = 3.325, P = 0.005) and APACHEE Ⅱ scores (OR = 4.831, P < 0.001) were all independent risk factors of senile severe pneumonia. Compared with the single prediction of APC, BNP and APACHE Ⅱ scores, the sensitivity, specificity, positive predictive value and negative predictive value of the combined prediction of severe pneumonia death in the elderly were significantly increased (89.42%, 81.61%, 84.72% and 86.03%).

APC, BNP and APACHEE Ⅱ scores change significantly in the progression of senile severe pneumonia, and were independent risk factors of senile severe pneumonia. The combination of the three indexes could significantly improve the prognostic predictive value.

To evaluate the diagnosis value of 3D microscopic ultrasound sonography for hepatitis B related liver cirrhosis (LC) and liver function decompensation.

Total of 270 patients were admitted to the Fourth People’s Hospital of Qinghai Province from January 2016 to December 2017, which were divided into chronic hepatitis B group (102 cases), compensatory LC group (84 cases) and decompensation LC group (84 cases). 3D microscopic ultrasound sonography, 2D shear waved elastography (2D-SWE) and blood routine examination were performed on each patient. Aspartate aminotransferase-to-platelet ratio index (APRI) were calculated. 3D microscopic ultrasound sonography scores, 2D-SWE scores and APRI were compared among the three groups, respectively. Receptor operation curves were built to compare the predictive values of 3D microscopic ultrasound sonography scores, 2D-SWE scores and APRI for LC and for liver function decompensation of LC patients.

The score of 3D microscopic ultrasound sonography and liver hardness among the three groups were significantly different (F = 313.52, 173.36; both P < 0.001). The area under receptor operation curve (AUROC) of 3D microscopic ultrasound sonography for predicting cirrhosis was 0.925 (95%CI: 0.885-0.965, P < 0.001); with the cut-off value of 11.5 scores, the sensitivity and specificity of 3D microscopic ultrasound sonography for predicting cirrhosis were 85.7% and 90.2%, respectively. The AUROC of 3D microscopic ultrasound sonography for predicting decompensated cirrhosis was 0.850 (95%CI: 0.795-0.905, P < 0.001). With the cut-off value of 16.5 scores, the sensitivity and specificity of 3D microscopic ultrasound sonography for predicting decompensated cirrhosis were 60.7% and 89.3%, respectively.

A quantitative diagnostic standard based on 3D microscopic ultrasound sonography was established in this study, which could be used in the auxiliary diagnosis of LC and liver function decompensation.

To investigate the clinical characteristics and psychological status of perianal and rectal condyloma acuminatum (CA) of men with human immunodeficiency virus (HIV) infection who had sex with men (MSM).

The clinical characteristics of 64 MSM patients with HIV infection and perianal rectal CA in Sichuan Intercontinental Hospital of Proctology and Gastroenterology from July 2015 to June 2017 were analyzed, and all patients received the Symptom Checklist (SCL-90) scale for psychological assessment.

Among the MSM patients with HIV infection, perianal and rectal CA mainly occured in patients of 18-45 years old (61 cases, 84.72%), and 47 cases (73.44%) had a college degree. There were 56 cases (87.50%) with classical CA and 8 cases (12.50%) with flat type CA. The positive rate of HPV detection was 81.25% (52 cases, 52/64), and HPV11 was the main subtype (50.00%, 26/52). SCL-90 scale psychological assessment showed that negative mental emotional patients were 59 cases (92.19%), mainly manifested in depression of 48 cases (75%) and anxiety of 7 cases (10.94%).

Young adults are the main crowd of perianal and rectal CA among HIV infected MSM patients. The most common type was classical CA, and their psychological status were mainly depression and anxiety.

To establish a relative quantitative assay for lactoferrin-DNA complex, and to assess the levels of neutrophil extracellular traps (NETs)in plasma samples from patients with severe influenza A (H1N1) and other pathogen infectious diseases.

Plasma samples with clear DNA content were serially diluted as a standard, and the level of lactoferrin-DNA complex in plasma was determined by Enzyme-linked immunosorbent assay (ELISA). The lactoferrin-DNA ELISA method was used to compare the difference of plasma lactoferrin-DNA complex between 19 patients with severe influenza A (H1N1) and 19 blood donors (control group).

Level of plasma lactoferrin-DNA complex of patients in severe influenza A (H1N1) was positively correlated with that of cfDNA (cell-free DNA) and histone-DNA complex, respectively (r = 0.6763, P < 0.0001; r = 0.756, P < 0.0001). The plasma lactoferrin-DNA complex content in patients with severe influenza A (H1N1) was 2.082 (1.169, 5.021) μg/ml, which was significantly higher than that of the controls [0.233 (0.170, 0.376) μg/ml, P < 0.0001], and positively correlated with Acute Physiological and Chronic Health Evaluation Ⅱ score (APACHE Ⅱ score) (r = 0.7379, P = 0.0005).

Plasma lactoferrin-DNA complex level could be used to assess NETs level in patients with severe influenza A (H1N1).

To investigate the changes and clinical significance of immune function of neonates with infectious pneumonia.

Total of 60 neonates with infectious pneumonia admitted in the Department of Neonatology, the Central Hospital of Longhua District from March 2016 to October 2017 were selected as observation group, while 60 healthy newborns were selected as the control group during the same period. According to the severity of pneumonia, cases in observation group were divided into mild group (39 cases) and severe group (21 cases). According to the staging difference of neonatal infectious pneumonia, cases in observation group were divided into acute stage group (24 cases) and convalescent stage group (36 cases). The blood samples of all children were collected in acute period and recovery period, for the detection of humoral immunity indexes (IgA, IgM, IgG1, IgG2, IgG3, IgG4 and complement C3, C4) and cell immunity indexes (CD3⁺ T, CD4⁺ T, CD8⁺ T, CD4/CD8, NK cells).

The serum levels of IgA, IgG1, IgG2, IgG3, IgG4, the ratios of CD3⁺ T cells, CD4⁺ T cells, NK cell and CD4/CD8 in cases of severe group were significantly lower than those of the control group and mild group (all P < 0.05, but the ratios of IgM, C3, C4 and CD8⁺ T cells were significantly higher than those of the control group and mild group (all P < 0.05). Spearman correlation analysis showed that the levels of serum IgA, IgG1, IgG2, IgG3, IgG4, the ratios of CD3⁺ T cells, CD4⁺ T cells, NK cells and CD4/CD8 were negatively correlated with the severity of neonatal infectious pneumonia (r =-0.826, -0.826, -0.665, -0.822, -0.826, -0.816, -0.794, -0.824, -0.820; all P < 0.001). The level of serum C3 and CD8⁺ T cell ratio were positively correlated with the severity of neonatal infectious pneumonia (r = 0.467, 0.788; all P < 0.001). The levels of serum IgA, IgG1, IgG2, IgG3, IgG4, CD3⁺ T cells, CD4⁺ T cells, NK cells and CD4/CD8 of cases in the acute stage were significantly lower than those of the convalescent stage group and the control group (all P < 0.05), but the levels of serum IgM, C3, C4 and CD8⁺ T cells were significantly higher than those of the convalescent stage group and the control group, all with significant differences (all P < 0.05).

Cellular immunity and humoral immunity function of patients with neonatal infectious pneumonia declined, which were negatively correlated with the severity of disease. Diseases at different stages have great influence on immune status, which should be focused on the regulation of immune status of these neonates.

To investigate the safety of different withdraw timepoint of tenofovir disoproxil (TDF) for pregnant women with high hepatitis B virus (HBV) DNA load in blocking mother-to-child transmission of HBV.

From January 2015 to December 2017, a total of 109 pregnant women with HBsAg positive and immunoresistance were recruited from Shenzhen Third People’s Hospital, all patients were treated with TDF at 24-28 weeks of gestation, who were divided into two groups according to simple randomized grouping method: withdrawal immediately after delivery group (58 cases) and withdrawal 4-12 weeks after delivery group (51 cases). The levels of HBV DNA and ALT were measured quantitatively at 4 weeks, 8 weeks, 12 weeks after antiviral therapy, at withdrawal and 4 weeks, 8 weeks, 12 weeks and 24 weeks after TDF discontinuation in both groups. Quantitative HBsAg and HBsAb levels of newborns at 4 weeks after birth were detected.

HBV DNA levels of all pregnant women before delivery were significantly lower than those of the baseline (withdrawal immediately after delivery group: Z = 8.459, P < 0.001; withdrawal 4-12 weeks after delivery group: Z = 7.760, P < 0.001). HBV DNA levels at the timepoint of withdrawal between the two groups were significantly different (Z = 2.242, P = 0.025). The difference of HBV DNA level 4 weeks after withdrawal between the two groups was not significant (Z = 1.041, P = 0.298), and no significant differences were found compared with the baseline levels (withdrawal immediately after delivery group: Z = 0.155, P = 0.877; withdrawal at 4-12 weeks after delivery group: Z = 0.376, P = 0.707). The difference of postpartum ALT levels between the two groups following up until 24 weeks after withdrawal was not significantly different (χ² = 1.319, P = 0.251). The median timepoint of ALT elevation of both groups was 4 weeks after withdrawal, with no significant difference (Z = 0.196, P = 0.844). The level of ALT increase during pregnancy may cause ALT increase after delivery, but it is not an independent risk factor. Total of 20 newborns were quantitatively detected for peripheral blood HBsAg and HBsAb at 4 weeks postpartum, all of them were HBsAg negative and successfully produced protective antibody.

TDF application in middle and late pregnancy could effectively block HBV transmission from mother to child. Following up for 24 weeks showed that there was no difference in postpartum safety between postpartum withdrawal and delayed withdrawal. Newborns produce protective antibody 4 weeks after birth and breast feeding was safe.

To evaluate the capabilities of disc diffusion and Vitek-2 Compact methods for testing antimicrobial susceptibility of Burkholderia cepacia.

Total of 34 isolates of Burkholderia cepacia were collected from January 2017 to December 2017 in Shaoxing People’s Hospital, the vitro minimum inhibition concentration (MIC) values of ceftazidime, compound sulfamethoxazole, levofloxacin, piperacillin/tazobactam, tetracycline, meropenem, chloramphenicol, minocycline and cefperidone/sulbactam were detected by disc diffusion method, Vitek-2 Compact method and broth microdilution method, respectively. Categorical agreement (CA) rates of disc diffusion and Vitek-2 Compact methods were determined, taking broth microdilution method as the reference method.

The 95% confidence intervals (95%CI) of ceftazidime, compound sulfamethoxazole, levofloxacin, meropenem and chloramphenicol were 6.2-13.2, 0.5-0.8, 3.8-6.3, 3.7-7.3 and 5.0-10.0, respectively, detected by broth microdilution method. The CA values of disc diffusion for ceftazidime, compound sulfamethoxazole, tigocycline, chloramphenicol and minocycline antibiotics were all > 90%, compared with broth microdilution method, Disc diffusion method and Vitek-2 Compact method were taken to detect the sensitivity of piperacillin/tazobactam in vitro, and the rate of agreement with reference method was the lowest, all < 80%. The major error (ME) of Vitek-2 Compact for detection of Tegafycline was 50%. Taken the broth microdilution method as the standard, the best antimicrobial agent in vitro was compound sulfamethoxazole and minocycline (all 100%), and the most sensitive antibacterials were levofloxacin (38.2%).

The drug sensitivity of Burkholderia cepacia detected by disk diffusion method and Vitek-2 Compact instrument method both had good reliability to ceftazidime and compound sulfamethoxazole. Ateention should be paid to the possible occurrence of very major error (VME) and ME, when antimicrobial susceptibility of piperacillin/tazobactam and cefoperazone/sulbactam were detected by disk diffusion method.

To evaluate the coagulation and platelet function in patients of acute coronary syndrome (ACS) complicated with HIV infection by thromboelastography.

From April 2015 to February 2018, a total of 32 patients of ACS complicated with HIV infection in Beijing Ditan Hospital, Capital Medical University were selected as observation group, while ACS patients with HIV negative were selected as the control group (32 cases). Thromboelastography, routine coagulation and platelet were taken to detect the coagulation and platelet function. The coagulation and platelet function indexes of the two groups were compared, including reaction time (R), coagulation time (K), coagulation angle (αangle), the maximal amplitude (MA), aspirin and clopidogrel resistance rate, the routine coagulation index and platelet count.

There was no significant difference in average age, hypertension, diabetes, hyperlipidemia, smoking and family history between the two groups (allP> 0.05). R, K were significantly lower in ACS patients with HIV infection than those of single ACS group (t =-4.00, 2.15;P < 0.01,P= 0.03);αangle was significantly higher of patients with ACS and HIV infection than that of single ACS group (t= 2.15,P= 0.03). But there was no significant difference in MA, ADP inhibiting rate and AA inhibiting rate between the two groups (allP> 0.05). There were no significant difference of the levels of PT, APPT, fibrinogen and platelet count of patients in ACS with HIV group than those of single ACS group (all P> 0.05). The reaction time was linearly correlated with APTT (r = 0.39,P = 0.04). The maximal amplitude was linearly related to the platelet count (r= 0.47,P= 0.01) and the content of fibrinogen of the patients (r= 0.68,P< 0.01).

Routine monitoring of thromboelastography in patients with HIV infection and acute coronary syndrome may provide more information on coagulation and platelet function. Patients with ACS and HIV infection are in hypercoagulable state, but adjustment of the antiplatelet therapy regimen due to the condition of HIV is not necessary.

To investigate the preventive effect of risk assessment model based on Logistic regression analysis on nosocomial infection.

The clinical data of 1 626 hospitalized patients in our hospital from January 2008 to December 2014 in the Center Hospital of Xiaogan Affiliated to Wuhan University of Science and Technology were analyzed, retrospectively, the patients were divided into the infection group (520 cases) and the control group (1 106 cases) according to whether with hospital infection during hospitalization. Logistic regression analysis was used to determine the risk factors of nosocomial infection and form a risk assessment scale. A total of 352 hospitalized patients admitted from January 2015 to February 2016 were assessed by this sacle for nosocomial infection risk.

Hospitalized time > 15 days, the application of triple antibiotics and ventilator, indwelling catheter, general anesthesia, liver disease, complicated with blood diseases, diabetes, hormone therapy, radiotherapy or chemotherapy, surgery time > 3 h, invasive operation were all independent risk factors for nosocomial infection (all P < 0.05). The incidence of nosocomial infection after establishment of the risk assessment scale was 25.00%, which was lower than that before the scale established (31.98%), with significant difference (χ² = 6.622, P < 0.05).

The risk assessment model based on Logistic regression analysis could effectively assess the risk of infection in patients, provide a basis for the prevention of nosocomial infections, and reduce the risk of nosocomial infections.

To investigate the value of clinical pulmonary infection score (CPIS) and procalcitonin (PCT) in the treatment of elderly patients with severe community-acquired pneumonia (SCAP).

Total of 60 elderly patients with SCAP in our hospital from February 2014 to February 2016 were collected. According to the prognosis, the patients were divided into survival group and death group. The difference of PCT, white blood cell (WBC), CPIS score between the two groups at the first day and 7th day after hospital admission were detected and their correlations with CPIS were analyzed.

Compared with the first day, the levels of PCT and CPIS in the survival group were not significantly decreased at the 7th day after hospital admission (P all < 0.001), but the WBC was without significant change (P > 0.05). Compared with the first day, the levels of WBC, PCT and CPIS in the death group were not significantly changed (P all > 0.05). After 7 days treatment, there were significant difference in PCT and CPIS between the two groups (P all < 0.05). WBC, PCT, and CPIS in SCAP patients had no significant correlation with prognosis on the first day (P all > 0.05). After 7 days treatment, WBC was not associated with prognosis (P > 0.05), and PCT and CPIS were significantly correlated with the prognosis of SCAP (r = 0.44, P = 0.023; r = 0.58, P = 0.017).

PCT and CPIS were associated with severity and prognosis in elderly patients with SCAP in elderly patients. It could be used as a good biomarker for evaluating the prognosis of SCAP, which could be used for further treatment.