Objective To develop the cDNA microarray for the combined detection of hepatitis virus, and to study the feasibility of applying the microarray in clinical setting. Methods For the small and simple genome of HBV and HDV, the specific primers of PCR were designed with Primer Premier 5.0 program according to the conserved region of HBV and HDV, and 10 and 4 gene fragments were obtained respectively, which could be used as the probes of gene chip. As for the complex genome of HCV, the technique of restriction display PCR (RD-PCR) was employed. Some of gene fragments were selected which were comparatively more specific and sensitive as microarray probes. In order to explore the experimental conditions of microarray in vitro detection, three types of gene chip were prepared successively including HBV and HDV simultaneous detection, HCV detection and modified HCV detection. Results The hybridized signals on the gene chip showed that the effect in detection was satisfactory. Through the prepared gene chips mentioned above, some probes with good quality were selected and the microarray was prepared for HBV, HCV and HDV simultaneous detection. The diagnostic capability of the microarray was evaluated following the washing and scanning steps. Linearity: Serial dilutions of the target DNA or cDNA showed that a strong linear relationship existed between the various concentrations of target DNA or cDNA and the fluorescence intensities obtained from microarray assay (r=0.990 2, r=0.992 1, r=0.981 9), and that the detection range for the microarray was from 104 to 1011 copies/ml. Specificity: Samples from other viruses such as YFV, JET and DV were also subjected to the test and the results were all negative. Reproducibility: The reproducibility of this assay system was evaluated by repeated measurements, and the within-run coefficient of validation of HBV, HCV and HDV were 7.1%, 7.2% and 6.6%, respectively, while the between-run coefficient of validation was 7.9%, 8.2% and 7.6%, respectively. Accuracy: By using the BLAST and the GenBank database, the identity of the obtained sequences including 16 fragments of PCR, 24 RD fragments of HCV and some positive serum samples were verified, each sequenced product was confirmed to be a genome fragment of expected size. In order to fulfill the request of clinical diagnosis, a modified protocol for microarray detection was established. This new protocol consisted of two hours of hybridization, omitting the steps of prehybridization and purification of samples, and the hybridization temperature was elevated from 42℃ to 52℃, et al. The whole protocol could be completed in less than 8 hours. 98, 42 and 5 serum samples from hepatitis B, C and D patients and 130 samples from healthy people were analyzed, respectively, by microarray assay and real-time PCR (Taqmam method). There was a significant correlation between the results of these assays (HBV, r=0.985 4 and HCV, r=0.958 2, P

ObjectiveTo evaluate whether AN69 ST membrane would prolong filter lifetime in continuous renal replacement therapy (CRRT) without anticoagulation in patients with high risk of bleeding.Methods A single-center, prospective, randomized, double-blind control trial with crossover design was conducted. From March 1st to December 31st in 2013, patients who were admitted to Department of Critical Care Medicine of the Fourth Hospital of Hebei Medical University meeting CRRT treatment indications, but could not receive systemic anticoagulation because of high risk of bleeding were studied. The selected patients were randomly divided into two groups according to a random number table, and four filters consisting of two AN69 ST100 membrane filters (A) and two traditional AN69 M100 membrane filters (B) were used for them. GroupⅠ with the filter order of A-B-A-B, and groupⅡ with the order of B-A-B-A. The clinical data of patients was recorded in detail, and conventional AN69 ST and AN69 membrane filter lifetime, their influence on coagulability, and the incidence of bleeding complications were compared.Results Seventeen patients were enrolled, with 10 in groupⅠ, and 7 in groupⅡ. The basic medical characteristics including gender, age, acute physiology and chronic health evaluationⅡ (APAECHⅡ) score, sequential organ failure score (SOFA), Acute Renal Injury Network (AKIN) stage, activated partial thromboplastin time (APTT), prothrombin time (PT), international normalized ratio (INR), platelet count (PLT), and use of mechanical ventilation were not significantly different between two groups. But the use of vasoactive drug was more frequent in groupⅡcompared with that of groupⅠ[100.0% (7/7) vs. 30.0% (3/10),χ2 = 8.330,P = 0.010]. AN69 ST filter lifetime (n =34) was (15.92±2.10) hours, there was no statistically significant difference compared with that of AN69 membrane (t = 0.088,P = 0.942), filter lifetime of which (n = 34) was (16.12±1.38) hours. It was also found by Kaplan-Meier survival analysis that there was no significant difference between the two membrane filter lifetime (χ2=1.589,P =0.208). Logistic regression analysis showed that the life of the first filter was not correlated with coagulation indicators, including APTT, PT, INR, and PLT [APTT: odds ratio (OR) = 0.977, 95% confidence interval (95%CI) = 0.892-1.071, P = 0.623; PT:OR = 1.001, 95%CI = 0.901-1.109,P = 0.988; INR:OR = 1.078, 95%CI = 0.348-3.340,P = 0.896;PLT:OR = 0.996, 95%CI = 0.974-1.019,P = 0.735]. The application rate of vasoactive drugs, which was different between two groups for basic medical indications showed no effect on filter life time (OR = 2.541, 95%CI = 0.239-26.955,P = 0.439). Reasons of clotting in filters were also analyzed, and it was found that blood coagulation in the filter ranked the top (88.2%), and the other reasons were catheter-related problems, death, and unscheduled transport. No difference in blood coagulation function was found in both groups after treatment for 12 hours, and there was no bleeding complication.ConclusionDuring the CRRT without systemic anticoagulant, both surface-treatment with polyethyleneimine AN69 and AN69 ST membrane cannot prolong filter lifetime.

Objective:To construct superparamagnetic iron oxide nanoparticles targeting tumor angiogenesis and evaluate their potential value as contrast agent in magnetic resonance imaging (MRI) .Methods:Ultrasmall superparamagnetic iron oxide (USPIO) nanoparticles targeting tumor angiogenesis were prepared by using co-precipitation chemical method. Cyclic RGD(cRGD) containing the sequence of Arg-Gly-Asp were conjugated USPIO nanoparticles by using chemical conjugative method to prepare superparamagnetic imaging agent targeting tumor angiogenic vessles. The physical and chemical properties of cRGD-USPIO nanoparticles were detected. The specific binding capabilities of cRGD-USPIO and USPIO to human lung adenocarcinoma cells (A549) and human umbilical vein endothelial cells (HUVEC) were tested by Prussian blue staining. A549 xenografts were established in nude mice, then USPIO and cRGD-USPIO were injected though tail vein, and the MRI signal enhancement effect of cRGD-USPIO was evaluated.Results:We successfully prepared the cRGD-USPIO nanoparticles. Its core diameter was 5-10 nm and the average diameter was (43.97±10.10) nm and the quality saturation magnetic intensity was 59.94 A·m~2·kg~(-1). Cell-binding test suggested that cRGD-USPIO group showed strengthened positive staining. In vivo MRI experiments showed that signals of tumor were significantly reduced in cRGD-USPIO group than that in USPIO group (P<0.01). Conclusion:The constructed cRGD-USPIO nanoparticles can be developed as a potential tumor-specific MRI contrast agent for the early diagnosis of cancer.

Objective To investigate The Th1/Th2 and Tc1/Tc2 polarization in the peripheral blood of first degree relatives of pemphigus vulgaris(PV) and healthy control individuals, and to approach the mechanism of the Dsg3-specific autoimmunity in PV. Methods The peripheral blood mononuclear cells (PBMC) from first degree relatives and healthy control was stimulated for72 h with Dsg3 and without Dsg3. Th1/Th2, Tc1/Tc2 was assessed by four-color flow cytometry. Results The mean frequency of Dsg3-spe-cific Th2 cells for PV antibody positive first degree relatives was 10.13%±3.72%, compared with stimula-tion without antigen 7.28%±3.58%, the difference was significant (P<0.05). The percentage Dsg3-spe-cific Th2 was markedly higher in the PV antibody positive first degree relatives group than that in the control group(10.13%±3.72% vs 6.10%±2.82%, P<0.05) , Tc2 was markedly higher also (20.01%± 10.43% v514.91%±8.06%, 20.01%±10.43% vs 9.58%±5.49%, P<0.05). Conclusion When Dsg3 stimulated PBMC were used to stimulate autologous T cells an increased amount of Th2 and Tc2 was observed, it is implied that the imbalance of Th1/Th2, Tc1/Tc2 might play an important role in the initia-tion of PV.

Objective To assess the sensitivity of indices of hip development in children with spastic diple-gia resulting from cerebral palsy. Methods X-ray images of the hips of 57 children with cerebral palsy ( the cere-bral palsy group) were checked, and the acetabular index ( AI), femur head migration percentage ( MP), center-edge angle and neck-shaft angle (NSA) were compared with those of normal children ( the control group, n = 30).Results The differences in MP and NSA between the two groups were significant. The prevalence of hip subluxation was 20.45% among the children with spastic diplegia who could not walk independently, and the prevalence was sig-nificantly greater in children 3 to 5 years old than among those under 3. Conclusion The MP can be used as a sen-sitive index to evaluate hip development. Age is a relevant factor affecting the hip development of children with cere-bral palsy.

Objective To investigate the associations of baseline serum uric acid, bilirubin levels with short-term outcome in patients with acute ischemic stroke. Methods The clinical data in successive patients with acute ischemic stroke were colected, including the serum levels of uric acid and bilirubin on admission, the National Institutes of Health Stroke Scale (NIHSS) score, and the modified Rankin scale (mRS) score at discharge or at day14 (mRS 0-2 was defined as good outcome, > 2 was defined as poor outcome). Results A total of 162 patients with ischemic stroke were enroled, including 114 in the good outcome group and 48 in the poor outcome group. There were significant differences in proportions of the patients with diabetes melitus (51. 75% vs. 75. 00% ; χ2 = 7. 526, P = 0. 006), previous history of stroke or transient ischemic attack (TIA) (18. 42% vs. 50. 00% ; χ2 = 17. 790, P = 0. 001), as wel as the baseline diastolic blood pressure (87. 061 ± 12. 245 mmHg vs. 82. 375 ± 10. 949 mmHg; t = 2. 293, P = 0. 023; 1 mmHg =0. 133 kPa), high-density lipoprotein cholesterol (1. 604 ± 0. 299 mmol/L vs. 1. 265 ± 0. 206 mmol/L; t =3. 227, P = 0. 002), fasting glucose (2. 875 ± 0. 438 mmol/L vs. 8. 160 ± 0. 592 mmol/L; t = - 4. 761, P <0. 001), uric acid (289. 365 ± 77. 168 μmol/L vs. 248. 206 ± 66. 206 μmol/L; t = 3. 111, P = 0. 002), total bilirubin (14. 673 ± 2. 213 μmol/L vs. 10. 395 ± 2. 714 μmol/L; t = 3. 779, P = 0. 001 ), direct bilirubin (6. 036 ± 1. 392 μmol/L vs. 4. 956 ± 1. 379 μmol/L; t = 2. 088, P = 0. 038), and indirect bilirubin (8. 634 ± 2. 307 μmol/L vs. 5. 439 ± 1. 223 μmol/L; t = 4. 219, P < 0. 001) levels between the 2 groups. Multivariate logistic regression analysis showed that the previous history of stroke or TIA (odds ratio [ OR ] 3. 751, 95% confidence interval [CI ] 1. 395-10. 091; P = 0. 009) and baseline NIHSS score (OR 2. 723, 95% CI 1. 093-6. 783; P = 0. 031) were the independent risk factors for poor outcome of ischemic stroke; while uric acid (OR 0. 357, 95% CI 0. 141-0. 900; P = 0. 029), high-density lipoprotein (OR 0. 262, 95% CI 0. 079-0. 870; P = 0. 029), and indirect bilirubin (OR 0. 117, 95% CI 0. 025-0. 539; P = 0. 006) were independently correlated with good outcome. Conclusions The increased baseline uric acid and indirect bilirubin levels are the favorable factors for good outcome in patients with acute ischemic stroke.

Objective To explore teaching effectsof biochemistry by comparingthe traditional teaching with the network investigative teaching. Methods 425 five-year-program students of clinical medicine of grade 2012 were chosen as subjects, with 246 students from Classes 5-8 as experimental group in which network investigative teaching was implemented andwith 179 students of Classes 17-20 as the control group which wasgiven traditional teaching. After the teaching , the teaching effects were evaluated through a questionnaire and the test scores of the two groups were compared and statistically analyzed. The two groups were compared by using t test. Results More than 70% of students in the experimental groups were generally satisfied with several aspects of the network investigative teaching and gavea good evaluation. The experimental group students' total scores and every item scores were all higher than the control group students', in which the practise scores and the case analysis problem scores had significant differences by statistical analysis with P values being 0.000 and 0.002, respec-tively. Conclusion Network investigative teaching is better than traditional teaching in the teaching of biochemistry , which can enhance students' ability of problem analysis and their enthusiasm for learning.

Objective:With Diagnosis related groups(DRG)indicator introduced into the hospital efficiency evaluation, to analyze the adjustment effect of DRG on the efficiency value, and to compare the similarities and differences between the DEA efficiency value and the DRG efficiency indicator.Methods:The DRG pilot data in 2017 and 2018 of tertiary hospitals in Hubei province and related data from other reports were collected. Indicators including the actual number of beds, the number of employees, the actual total bed days occupied, and medical income were used as input indicators, and total admissions, number of discharges, weight of DRG and number of DRG groups were used as output indicators. 7 models were constructed using different indicator combinations. The efficiency of the hospital was calculated by the Bootstrap-DEA efficiency evaluation method. The relationship between hospital attributes and efficiency was analyzed by a generalized linear model.Results:There was an abnormal phenomenon that the higher the hospital level, the lower the efficiency in the efficiency value calculated by the traditional method. The efficiency values of county hospitals, city hospitals and provincial hospitals were 0.83, 0.74 and 0.71, respectively( P<0.01). DRG weight and group number were used as output for DEA analysis, and the efficiency values of county hospitals, city hospitals, and provincial hospitals were 0.95, 0.95 and 0.96( P=0.20)respectively. The DRG efficiency indications of county hospitals, city hospitals and provincial hospitals were 1.42, 1.11, 1.00 respectively( P<0.01). The higher the level, the lower the efficiency, while the efficiency values calculated by DEA were 0.84, 0.82, 0.86, respectively( P=0.58). Conclusions:The efficiency value calculated by the traditional method presents a systematic bias, which could be corrected effectively when DEA analysis using DRG weights is used as output indicator. The results developed new ideas of efficiency analysis for hospitals practicing DRG management.

Objective:To quantitatively measure the level of hospital human resources management of public hospitals in China with D-CHMS.Methods:From January through December of 2019, the Development Chinese Hospital Management Survey(D-CHMS)was used in field survey of relevant personnel of 36 tertiary general hospitals and 27 secondary general hospitals, with 10 persons chosen from each hospital. Survey data were subject to descriptive analysis and t test, for quantitative measurement of the human resources management level of these sample hospitals. Results:The level of human resources management in Chinese public hospitals was on the low side, as seven secondary dimensions scored 2.94 in average. To name a few, talents retention and talents recruitment scored the lowest, being (2.13±0.29) and (2.90±0.63) respectively. Tertiary hospitals presented an obviously better performance than secondary hospitals in incentives for best performers, talents retention and recruitment. Human resources management level of hospitals in China′s west regions scored the highest, followed by those in the east regions.Conclusions:Public hospitals in China, especially secondary hospitals were expected to enhance their human resources management. Approaches recommended include building a talent pool, developing a series of preferential policies, building development platforms, optimizing performance appraisal, motivating remuneration schemes, and offering unique employee value propositions, which serve to recruit and retain talents.

Objective:To investigate the impact of human epidermal growth factor receptor 2 (HER2) and C-reactive protein/albumin ratio (CAR) on the long-term survival of gastric cancer patients after surgery.Methods:A retrospective analysis was conducted on 136 gastric cancer patients admitted to Lishui Central Hospital from January 2015 to December 2017, who underwent radical surgery and were followed up for 5 years. Patients were divided into HER2 positive and HER2 negative groups based on HER2 immunohistochemical results, and into high CAR and low CAR groups based on the CAR mean value. The relationship between HER2 and CAR with clinicopathological characteristics of gastric cancer patients was analyzed. The postoperative tumor-free survival rate and overall survival rate were compared between the two groups of patients (HER2 positive group and HER2 negative group, as well as high CAR group and low CAR group). Cox regression analysis was used to identify independent prognostic factors for postoperative tumor recurrence, metastasis, and death in gastric cancer patients.Results:The proportion of HER2 positive patients with large tumor size, low differentiation, T 3-4 tumor invasion depth, lymph node metastasis, and vascular invasion was significantly higher than that of HER2 negative patients (all P<0.05). The proportion of high CAR patients with large tumor size, low differentiation, T 3-4 tumor invasion depth, lymph node metastasis, and vascular invasion was significantly higher than that of low CAR patients (all P<0.05). Kaplan-Meier survival analysis showed that HER2 negative patients had significantly higher 1-year, 3-year, and 5-year cumulative tumor-free survival rate and overall survival rate than HER2 positive patients, while low CAR patients had significantly higher 1-year, 3-year, and 5-year cumulative tumor-free survival rate and overall survival rate than high CAR patients (all P<0.05). Multivariate Cox regression analysis identified T 3-4 tumor invasion depth, lymph node metastasis, HER2 positivity, and high CAR expression as independent prognostic factors for postoperative tumor recurrence, metastasis, and death in gastric cancer patients (all P<0.05). HER2 positive gastric cancer patients had a 1.895-fold higher risk of postoperative tumor recurrence and metastasis than HER2 negative patients ( HR: 1.895, 95% CI: 1.245-4.229, P=0.034), while high CAR gastric cancer patients had a 1.769-fold higher risk of postoperative tumor recurrence and metastasis than low CAR patients ( HR: 1.769, 95% CI: 1.433-3.959, P=0.039). HER2 positive gastric cancer patients had a 2.145-fold higher risk of postoperative death than HER2 negative patients ( HR: 2.145, 95% CI: 1.378-4.589, P=0.028), while high CAR gastric cancer patients had a 1.926-fold higher risk of postoperative death than low CAR patients ( HR: 1.926, 95% CI: 1.564-3.853, P=0.025). Conclusions:HER2 and CAR are independent prognostic factors for postoperative tumor recurrence, metastasis, and death in gastric cancer patients. Gastric cancer patients with HER2 positivity and high CAR have a higher risk of postoperative tumor recurrence, metastasis, and death. This study has some limitations due to its small sample size and single-center design, which may introduce some bias. Future multicenter and large-scale studies are needed to confirm the results of this study.