Molecular characterisation and expression profile of the PRLR gene during goose ovarian follicle development.

1. Although PRL-PRLR signalling plays important roles in regulating avian reproduction, there is a paucity of information regarding the functional significance of PRLR in goose ovarian follicle development.2. The full-length 2,496 bp coding sequence of PRLR was obtained from Sichuan White goose (Anser cygnoides) for the first time and was seen to encode a polypeptide containing 831 amino acids. Goose PRLR shares similar sequence characteristics and conserved functional domains to other avian species and was phylogenetically clustered into the avian clade.3. The qPCR results suggested that the mRNA levels of PRLR significantly increased in primary follicles during weeks 3 to 4 of age and were higher in secondary- than in primordial follicles at week 5 post-hatching, which suggested that the PRLR-mediated signalling could be involved in regulation of early folliculogenesis.4. The PRLR mRNA was expressed at the highest levels in the prehierarchical 8-10 mm granulosa layers throughout goose ovarian follicle development, indicating a role for PRLR in the process of follicle selection.5. PRLR mRNA was differentially expressed in the three cohorts of in vitro cultured granulosa cells harvested from different sized goose ovarian follicles, which suggested that PRLR was involved in regulating granulosa cell functions depending on the stage of follicle development. These data provide novel insights into the role of PRLR during goose ovarian follicle development, although the underlying mechanisms await further investigations.

[The expression and clinical significance of long non-coding RNA GIHCG in cholangiocarcinoma].

Objective: To screen long non-coding RNA (lncRNA) related to the prognosis of cholangiocarcinoma patients, detect its expression in cholangiocarcinoma tissue, and analyze its clinical significance by analyzing The Cancer Genome Atlas (TCGA) database. Methods: Using limma package, survival package, and survival receiver operating characteristic curve (ROC) package of R software to analyze the data of cholangiocarcinoma in TCGA and screen the differentially expressed lncRNAs related to patient survival. Real-time PCR and Fish were used to detect the expression of lncRNA and analyze its correlation with the clinical characteristics of patients. Small interfering RNA was used to knock down the expression of lncRNA GIHCG, and its effect on the migration ability of cholangiocarcinoma cell lines was detected by Transwell. Results: The results of the comprehensive analysis of survival, ROC, and correlation analysis with clinical data showed that lncRNA GIHCG has a significant correlation with lymph node metastasis in patients with cholangiocarcinoma. The expression of lncRNA GIHCG in cholangiocarcinoma tissue is significantly increased, closely related to tumor size and lymph node metastasis. Transwell results showed that lncRNA GIHCG could promote the migration of cholangiocarcinoma cells. Conclusion: The expression of lncRNA GIHCG is significantly increased in cholangiocarcinoma tissues and is closely related to patient survival and lymph node metastasis. It is expected to become a new molecular marker for diagnosing or treating cholangiocarcinoma.

Research progress into the physiological changes in metabolic pathways in waterfowl with hepatic steatosis.

1. When geese or ducks are overfed with a high-energy diet rich in carbohydrates, their liver increases in size by 5- to 10-fold in 2 weeks, which is accompanied by the occurrence of hepatic steatosis. As a result, this distinctive genetic characteristic of waterfowl has been taken advantage of to produce foie gras. 2. When overfed geese were fed a regular diet for a 20-d period of recovery, their liver was restored to the original state. Hence, the entire process is reversible, and cause no lasting cirrhosis or necrosis in the liver. This suggests that waterfowl have a mechanism to protect their liver from the harm caused by severe hepatic steatosis. 3. This paper reviews the formation, physiological changes to metabolic pathways and the protective mechanisms in geese and ducks with hepatic steatosis. Not only will understanding these mechanisms provide ideas for the waterfowl breeding selection for foie gras, it is conducive to improving production efficiency and quality of foie gras. This provides a scientific basis to ensure animal welfare and an approach to the prevention and treatment of fatty liver disease in human.

[The safety and efficacy of combined hepatic artery resection in treatment of hilar cholangiocarcinoma: a meta-analysis].

Objective: To evaluate the safety and effectiveness of combined hepatic artery resection for the treatment of hilar cholangiocarcinoma. Methods: We searched Pubmed, The Cochrane Library, Embase, Web of Science, China Knowledge Network, Wanfang Data Resource System, Vip-Chinese Sci-tech Journal System Database, and China Biomedical Literature Database, and collected the randomized controlled studies or retrospective studies on the safety and efficacy of combined hepatic artery resection and non-hepatic artery resection in the treatment of hilar cholangiocarcinoma. The search period is from January 1, 2006 to December 31, 2019. Review Manager 5.3 software was used to analyze the extracted data indicators. Results: A total of 14 articles were collected, and a total of 2 374 patients with hilar cholangiocarcinoma were included in the study. Meta-analysis results showed that the perioperative mortality in the hepatic artery resection (HAR) group was higher than that of the control group (OR=1.70, 95%CI=0.02-2.90, P=0.05), and the total postoperative morbidity rate was higher than that of the control group (OR=1.28, 95%CI= 0.93-1.76, P=0.13), both of which were not statistically significant compared with the control group. Subgroup analysis showed that the incidence of liver failure (OR=1.15, 95%CI= 0.73-1.82, P=0.54), biliary fistula (OR=1.20, 95%CI= 0.78-1.84, P=0.40), and abdominal infection in the two groups (OR=0.98, 95%CI= 0.53-1.83, P=0.95) was without significant difference. The R0 resection rate of the HAR group was higher than that of the control group, and the difference was not statistically significant (OR=1.08, 95%CI=0.66-1.75, P=0.77). The rates of lymph node metastasis in the HAR group were higher than that in the control group (OR= 2.48, 95%CI= 1.05-5.84, P=0.04). One-year(OR=0.48, 95%CI= 0.32-0.72, P=0.000 5), 3-year (OR= 0.51, 95%CI=0.36-0.72, P=0.000 1), and 5-year (OR=0.50, 95%CI=0.35-0.70, P<0.000 1) survival rates of HAR group were lower than those of the control group. The survival rates of patients in HAR group treated with combined chemotherapy drugs after operation were significantly improved (OR= 7.33, P=0.02). Conclusions: The safety of combined HAR treatment for hilar cholangiocarcinoma is acceptable, but poor postoperative survival may be related to the high lymph node metastasis rate. Therefore, it is still necessary to be cautious in carrying out this operation. Combined with adjuvant chemotherapy after surgery may improve survival.

Effects of human umbilical cord mesenchymal stem cells on inflammatory factors and miR-181a in T lymphocytes from patients with systemic lupus erythematosus.

OBJECTIVES: The present study aimed to explore the effect of umbilical cord mesenchymal stem cells (UC-MSCs) on the modulation of T lymphocytes from system lupus erythematosus (SLE) patients and the possible mechanism. METHODS: A total of 24 hospitalized SLE patients and 28 healthy individuals were enrolled. T lymphocytes were sorted using Miltenyi magnetic beads. After the addition of recombinant human interleukin (IL)-2 and CD3CD28 T-cell activator, cells were loaded onto six-well plates pre-inoculated or not with UC-MSCs for 1 week of culture. The supernatants were collected for testing inflammatory factors by enzyme-linked immunosorbent assay. Meanwhile, T lymphocytes were collected to assess the expression levels of genes, proteins in relation to SLE and miR-181a by polymerase chain reaction and Western blot. RESULTS: Compared with T lymphocytes cultured alone, interferon-γ, IL-4, IL-6 and IL-10 levels were significantly decreased in T lymphocytes from SLE patients co-cultured with UC-MSCs. In addition, the gene and protein expression levels of TNF alpha, osteopontin and nuclear factor-kappa B in T lymphocytes were significantly decreased, while miR-181a expression was markedly elevated (p < 0.05 or 0.008). CONCLUSION: UC-MSCs have showed certain immunomodulatory and inhibitory effects in vitro on T lymphocytes from SLE patients, which could potentially be a beneficial treatment of the disease. UC-MSCs may up-regulate miR-181a and down-regulate inflammation-related gene expression.

[Risk factors for the development of liver cancer in patients with hepatitis B-related liver cirrhosis treated with long-term nucleos(t)ide analogues].

Objective: To retrospectively analyze the risk factors for the development of liver cancer in patients with hepatitis B-related liver cirrhosis (LC) treated and fully managed with long-term nucleos(t)ide analogues (NAs). Methods: The study subjects were derived from the follow-up cohort of chronic hepatitis B and liver cirrhosis who received antiviral therapy in the Department of Infectious Diseases of the First Affiliated Hospital of Guangxi Medical University from February 2004 to September 2019. LC patients who met the inclusion criteria were enrolled. The life-table method was used to calculate the incidence of liver cancer. Multivariable Cox regression model was used to analyze the risk factors that may affect the development of liver cancer in patients with LC. A subgroup analysis was conducted in liver cirrhotic patients who developed liver cancer to evaluate the effectiveness of antiviral treatment compliance. The (2) test was used for rate comparison. Results: The median follow-up time of 198 LC cases treated with NAs was 6.0 years (1.0-15.3 years). By the end of the visit: (1) 16.2% (32/198) of LC patients had developed liver cancer, and the cumulative incidence of liver cancer in 1, 3, 5, 7, and 9 years were 0, 8.9%, 14.3%, 18.6%, and 23.4%, respectively, with an average annual incidence of 3.1%. Among the 32 cases with liver cancer, 68.7% had developed small liver cancer (22/32). (2) Univariate Cox model analysis showed that the development of liver cancer was related to four factors, i.e., the presence or absence of LC nodules, whether the baseline was first-line medication, the family history of liver cancer, and patient compliance. The results of multivariate Cox model analysis showed that poor patient compliance and baseline non-first-line medication were risk factors for liver cancer. (3) The results of log-rank test subgroup analysis showed that the 5-year cumulative incidence of liver cancer in patients with hardened nodules was significantly higher than that of patients without hardened nodules (21.7% vs. 11.5%, P = 0.029). The 5-year cumulative incidence of liver cancer in patients with non-first-line drugs was significantly higher than that of patients with first-line drugs (22.0% vs.8.2%, P = 0.003). The 5-year cumulative incidence of liver cancer in patients with poor compliance was significantly higher than that of patients with good compliance (21.3% vs. 12.7%, P = 0.014). The 5-year cumulative incidence of liver cancer in patients with a family history of liver cancer was significantly higher than that of patients without a family history of liver cancer (22.3% vs. 8.1%, P = 0.006). (4) Compared with patients with poor compliance, patients with good compliance had higher HBV DNA negative serconversion rate (98.7% vs. 87.8%, P = 0.005), and a lower virological breakthrough rate (12.1% vs. 29.3%, P = 0.007). Conclusion: The long-term NAs antiviral therapy can reduce the risk of liver cancer, but it cannot completely prevent the development of liver cancer, especially in patients with a family history of liver cancer and baseline hardened nodules (high risk of liver cancer). Furthermore, the complete management can improve patient compliance, ensure the efficacy of antiviral therapy, and reduce the risk of liver cancer development, so to achieve secondary prevention of liver cancer, i.e., early detection, diagnosis and treatment.

[Liver histological status and clinic outcome in HBeAg-negative chronic hepatitis B with low viral load].

Objective: To retrospectively analyze the serological, virological, biochemical, liver histological status and clinical outcomes in HBeAg-negative chronic hepatitis B (CHB) patients with low HBV viral load, and to explore the necessity of antiviral therapy for these patients. Methods: A total of 99 HBeAg-negative CHB patients with HBV DNA level < 4 lg copies/ml who performed liver biopsy at the baseline were enrolled from the follow-up cohort. Among them, 23 cases received the second liver biopsy during follow-up. The relationships among the degree of inflammation and fibrosis of liver tissues, the status of HBsAg and HBcAg, age, gender, family history, HBV DNA load, serological markers and other indicators were analyzed. The pathological differences between two liver biopsy examinations were compared. The effect of nucleos(t)ide analogues (NAs) treatment on patient's clinical outcomes were analyzed. For multivariate analysis, a binary logistic regression model was performed. Log-rank test was used to compare the cumulative incidence of hepatocellular carcinoma (HCC) in NAs-treated and non-NA streated patients. Results: Baseline liver histology status showed that 58.6% (58/99) patients had obvious liver tissue damage in their baseline liver tissue pathology (G≥2 and /or S≥2). Univariate logistic regression analysis showed that a liver cirrhosis (LC) family history, a HBsAg-positive family history, baseline alanine aminotransferase and aspartate aminotransferase levels were positively correlated factors for liver tissue damage. Multivariate logistic regression analysis showed that a LC family history was the main risk factor for liver tissue damage. Twenty-three cases had received a second liver biopsy after an interval of 4.5 years. In 10 untreated cases, the second liver biopsy results showed the rate of obvious liver tissue damage (G≥2 and/ or S≥2) increased from 50.0% to 90.0%. In the other 13 cases who received NAs treatment, the second liver biopsy showed improvement in liver histology, and the rate of obvious liver tissue damage decreased from 61.5% to 46.2%. The 5-year HCC cumulative incidence in non-NAs-treated patients was significantly higher than that of in NAs-treated patients (17.7% vs. 3.8%, P = 0.046). Conclusion: For most HBeAg-negative CHB patients with low viral load, liver tissue pathology result suggests that it meets the indications for antiviral therapy, especially in patients with a LC familial history. Without antiviral therapy, liver tissue damage for these patients will progressively worse with the high incidence of HCC. Therefore, it is suggested that antiviral therapy should be started as soon as possible for the HBeAg-negative CHB patients with low viral load regardless of the alanine aminotransferase level, especially in patients over 30 years-old with a LC or HCC family history.

[The effect of estrogen level on Budd Chiari syndrome related hepatocellular carcinoma].

Objective: To investigate the effect of estrogen level on Budd Chiari syndrome related hepatocellular carcinoma. Methods: Immunohistochemical method was used to detect estrogen receptor-α and estrogen receptor-ß expression in 38 cases of Budd Chiari syndrome related hepatocellular carcinoma and 50 cases of HBV related hepatocellular carcinoma.Hepatoma cells of Budd Chiari syndrome related hepatocellular carcinoma were exposed to different concentrations of Estrogen for 48 hours. Tetrazolium bromide (MTT) colorimetry was used to analyze cell proliferation activities; cell cycle was analyzed by flow cytometry (FCM); cell apoptosis was analyzed by flow cytometry (FCM) and Casepase-3 activity was measured after induced by adriamycin(ADM). Results: The positive rate of estrogen receptor-α expression in the tissues of Budd Chiari syndrome related hepatocellular carcinoma was 71.05%, which was higher than that (32%)in HBV related hepatocellular carcinoma tissue evidently (P<0.01). The positive rate of estrogen receptor-ß expression in the tissues of Budd Chiari syndrome related hepatocellular carcinoma was 68.4%, which was higher than that (26%)in HBV related hepatocellular carcinoma tissue evidently (P<0.01). With the concentrations of estrogen increasing, MTT Assays showed that estrogen level increased the cell proliferation activities of Budd Chiari syndrome related hepatocellular carcinoma. The number of cells at stage S and G2/M were significantly increased and cells at stage G0/G1 were reduced with the increasing concentrations of estrogen. After being incubated under the different concentrations of estrogen for 48 h, the apoptosis rates decreased gradually and the Casepase-3 activity was significantly reduced with the increasing concentration of estrogen. Conclusions: Estrogenreceptor expression may have an important influence on hepatocellular carcinoma cell biology difference between Budd Chiari syndrome related hepatocellular carcinoma and HBV related hepatocellular carcinoma. Estrogen level can promote cell proliferation and cell cycle, and inhibit the apoptosis of hepatoma cells of Budd Chiari syndrome related hepatocellular carcinoma in vitro, and these effects were increased with the increasing of estrogen level.

Characteristic Changes and 3D Virtual Measurement of Lung CT Image Parameters in the Drowning Rabbit Model.

OBJECTIVES: To use virtual anatomy technique in the analysis of post-mortem characteristic changes of CT images in the experimental drowning rabbit model and the related parameters in 3D virtual model, so as to explore its application value in the diagnosis of drowning in forensic pathology. METHODS: A model of drowning rabbits was established, with animal models of hemorrhagic shock and mechanical asphyxia as the controls. CT scan was performed on the experimental animals, and the differences in imaging features between the groups were compared by morphological reading of the tomographic images. CT data were imported into Mimics 14.0 software for 3D modeling. The CT values and lung volumes were calculated by the software, and the differences on CT values and lung volumes brought by different causes of death were analyzed. RESULTS: The CT images of lungs in the drowning group showed characteristic ground-glass opacity （diffuse and uniform density increase）. There were no obvious abnormalities in hemorrhagic shock group, and only a few similar changes were found in the mechanical asphyxia group. Compared with the controls, the CT values and the lung volumes in the drowning group were significantly increased P<0.05. CONCLUSIONS: Based on post-mortem lung imaging, the combination of CT value and lung volume changes can effectively reflect the virtual anatomical features in drowning, and provide a diagnostic basis for the forensic identification of drowning.

CHILD syndrome mimicking verrucous nevus in a Chinese patient responded well to the topical therapy of compound of simvastatin and cholesterol.

BACKGROUND: Congenital hemidysplasia with ichthyosiform erythroderma and limb defects (CHILD) syndrome is a rare X-linked dominant disorder characterized by peculiar cutaneous presentations and ipsilateral skeletal abnormalities. CHILD syndrome is caused by mutations in NSDHL gene, which involves in cholesterol synthesis. OBJECTIVES: To verify the diagnosis of CHILD syndrome and seek effective pathogenesis-based therapy with little side-effects. METHOD: We comprehensively evaluated the patient's conditions. Pathological biopsy was performed in the lesion location. Genetic tests and real-time quantitative PCR were conducted to further confirm the diagnosis. The topical application of a mixed lotion containing 2% simvastatin and 2% cholesterol to lesion areas based on the pathogenesis as well as the literature review. RESULTS: We diagnosed a rare and typical case of CHILD syndrome co-occurring with multiple VX-like lesions. The gene mutation is a large deletion of exon 3 and 4 of the NSDHL gene, which was discovered and reported for the first time in CHILD syndrome. The skin lesions, including the verruciform plaques and VX-like lesions, improved obviously after treatment. CONCLUSIONS: Multiple exons deletions or microdeletion was not rare in CHILD syndrome. Classical Sanger sequencing may not be useful enough to find all kinds of mutations. Next-generation sequencing may be more effective. It is important to conduct genetic counselling to prevent more serious defects in descendants. The excellent therapeutic effect on CHILD syndrome resulted from the topical treatment with simvastatin/cholesterol provides a proof-of-concept for other topical pathogenesis-based therapies for skin disease.

[Monitoring by high-sensitivity HBV DNA assay during treatment in chronic hepatitis B e antigen negative patients].

Objective: To explore the efficacy of tenofovir disoproxil and adefovir dipivoxil treatment in patients with hepatitis B virus e antigen (HBeAg) negative was analyzed through the comparison of highly sensitive HBV viral load monitoring with HBV genotyping and drug resistance mutations. Methods: The clinical data of newly diagnosed chronic hepatitis B patients from January 2015 to June 2017 in outpatients and inpatients were randomly divided into tenofovir and adefovir group. Quantitative detection of HBV DNA levels before therapy and at 12, 24, 48, 96, and 120 weeks after therapy were determined for HBV genotypes and drug-resistant mutations in HBeAg-negative patients. Student's t-test was used to compare the measurement data between groups. The data of comparison between groups were tested by χ (2). Results: A total of 106 cases of HBeAg-negative patients were collected. Tenofovir disoproxil had a higher rate of HBV DNA suppression (54%) than adefovir dipivoxil treatment (42%), but the difference was not statistically significant (P = 0.19). After 120 weeks of treatment, a total of 46 patients (93.9%) were enrolled in the tenofovir disoproxil group with HBV DNA quantitation < 2 000 IU / ml. Adefovir dipivoxil group of patients with HBV DNA < 2 000 IU / ml a total of 40 cases, accounting for 75.5%. The difference between the two groups was statistically significant (P < 0.05). For 49 cases of HBeAg-negative patients, HBV B, C, B and C were mixed before tenofovir dipivoxil treatment, and C1653T, A1762T and G1764A mutation sites were detected in patients with D genotype. Patients C, B, C, B, and C were examined for C1673T, G1896, G1858, G1899A. After treatment, the detection rate of the above mutation sites decreased, but C1653T, C1673T and G1899A were not detected. New mutation sites such as G1915A / C, L180M, M204V, V207I / L, T184A and V173L were detected, Low resistance rate (25%). Conclusion: Tenofovir disoproxil can be recommended as a treatment for HBeAg-negative patients. For HBeAg-negative patients, the choice of high-sensitivity detection of HBV DNA levels, better monitoring of anti-HBV efficacy.

Bifidobacterium breve as a delivery vector of IL-24 gene therapy for head and neck squamous cell carcinoma in vivo.

Beneficial bacteria are becoming ever more popular gene delivery method for hypoxia-tumor targeting in vivo. In this study we investigated the therapeutic effect of new recombinant Bifidobacterium breve strain expressing interleukin (IL)-24 gene (B. breve-IL24) on head and neck tumor xenograft in mice. Briefly, B. breve transformants were obtained through electro-transformation. Bacteria-tumor-targeting ability were analyzed in vivo over different time points (1, 3 and 7 days post-bacteria injection). Furthermore, the therapeutic effect of bacteria on tumor cells in vivo were analyzed as follows: 30 Balb/c nude mice bearing subcutaneous tumor were randomly divided in three groups (Drug group, green fluorescent protein (GFP) group and Saline group). The therapy lasted for 2 weeks and included B. breve-IL24 administration via tail vein for Drug group, B. breve-GFP for GFP group and phosphate buffered saline for Saline group. The tumor growth was monitored using standard caliper technique, while the apoptosis induction in vivo was analyzed by Real-time Positron Emission Tomography/Computed Tomography (PET/CT) imaging ([18F]-ML-10 tracer). At the end of the experiment, tumor tissues were collected and analyzed by western blotting. Briefly, our results suggested that our new recombinant bacterium has the capability of targeting tumor tissue in vivo. As for the therapeutic effect, our new strain has revealed to be a promising therapeutic approach against tumor growth in vivo. Briefly, higher tumor growth inhibition and higher tumor cell apoptosis induction were observed in Drug group compared with the GFP and Saline groups. To conclude, a new recombinant strain B. breve-IL24 offers a novel, safe and clinically acceptable therapeutic approach for tumor therapy in vivo.

Coupling metagenomics with cultivation to select host-specific probiotic micro-organisms for subtropical aquaculture.

AIMS: To demonstrate a nonempirical workflow to select host-specific probiotics for aquaculture industry. METHODS AND RESULTS: Using both culture-dependent and culture-independent methods, we have systematically investigated, for the first time, the gut microbiota of twelve subtropical aquatic animal species. We found that the diversity, abundance and distribution of gut micro-organisms of these animals were host-specific and that lactic acid bacteria (LAB) were predominant among the indigenous probiotic microbes. Using culturing method, we isolated and characterized ninety-eight LAB strains; however, only a few strains was representative of the dominant LAB OTUs recovered by culture-independent analysis. CONCLUSIONS: Two cultured LAB strains, Enterococcus faecalis LS1-2 and Enterococcus faecium Z1-2, capturing the major LAB OTUs in the sequencing data set of the most animal samples and showing significant antimicrobial activities against shrimp pathogens, were suggested to be the candidates of shrimp probiotics. SIGNIFICANCE AND IMPACT OF STUDY: Disease outbreak and the consequential abuse of antibiotics have been the constraints to the aquaculture industry. However, the selection of probiotic bacteria is currently still an empirical process due to our limited knowledge on the gastrointestinal microbiota of aquatic organisms. Our study points to a nonempirical selection process by which host-specific probiotics can be developed.

A practice of ultra-fine tailings disposal as filling material in a gold mine.

A practice of cemented backfill technology with ultra-fine tailings in a gold mine was comprehensively presented, and a series of tests were conducted in accordance with the peculiar properties of ultra-fine tailings and the mining technology conditions. The test results show that, the tailings from Shuiyindong Gold Mine have a great grinding fineness, with the average particle diameter 22.03 µm, in which the ultra-fine particles with the diameter below 20 µm occupying 66.13%. The analysis results of chemical components of tailings indicate that the content of SiO2 is relatively low, i.e., 33.08%, but the total content of CaO, MgO and Al2O3 is relatively high i.e., 36.5%. After the settlement of 4-6 h, the tailing slurry with the initial concentration of 40% has the maximum settling concentration of 54.692%, and the corresponding maximum settling unit weight is 1.497 g/cm3. During the field application, the ultra-fine tailings and PC32.5 cement were mixed with the cement-tailings ratios of 1:3-1:8, and the slurry concentration of 50 wt% was prepared. Using the slurry pump, the prepared cemented backfill slurries flowed into the goaf, and then the strength of the cemented backfill body met the mining technique requirements in Shuiyindong Gold Mine, where the ore body has a smooth occurrence, with the average thickness of approximately 2 m and the inclination angle ranging from 5 to 10°.

Effect of students' determination of testing time on their test performance.

This article investigates whether giving students control over preparing for and the moment of taking a test affects their test results in comparison with when the school is in control of the amount of training followed by a predetermined test moment. The students participated in training for manual dexterity. After the training, the students performed a test. The results of the test were stored in a database. Students from the group with freedom to select the moment of the test performed much better than those in the other group who did not have the freedom to select the moment for the test, with significantly fewer students requiring three attempts to pass the test. The fact that students when given the responsibility to develop manual skills performed better than when guided by the policy of the school is hopeful in the sense that students can learn in an early stage of their study to take responsibility for learning.

[Association between A-kinase anchor proteins 12 methylation and recurrence of hepatocellular carcinoma].

Objective: To study the association between the AKAP12 promoter methylation and recurrence of hepatocellular carcinoma. Methods: A total of 142 primary liver cancer patients underwent surgery in department of Hepatobiliary surgery in Peking University Cancer Hospital from 2003 to 2009 were selected as subjects in the survey; with the inclusion criteria as hepatocellular carcinoma, no cancer cells were observed in the surgical margin(SM) samples. All patients had neither lymph nor distant metastasis at the time of surgery, and receiving complete follow-up data for at least 3 years. By the end of May 2014, a total of 75 patients had relapsed of whom 71 died and there were no lost. All samples were acquired from the frozen surgical tissues. Genomic DNA was extracted using phenol/chloroform method and performed bisulfite modification following with polymerase chain reaction (PCR). AKAP12 methylation in hepatoma and the corresponding SM samples from 142 patients was determined by denature high-performance liquid chromatography (DHPLC) and bisulfite clone sequencing. Kaplan-Meier and Cox proportion hazard regression model were used to identify the factors related to the survival time. Results: In 142 cases, 125 patients (88.0%) were male and 17 (12.0%) cases were female. The median age was 52.5 years, ranging from 34 years to 76 years. AKAP12 methylation-positive rate was significantly higher in hepatomas than SMs (54.9% vs. 10.2%, P<0.001). Patients with AKAP12 methylation-positive had less risk of the recurrence (HR=0.62, 95%CI: 0.39-0.99); with tumor diameter more than 5 cm (HR=1.53, 95%CI: 1.00-2.50),portal vein invasion(HR=4.53, 95% CI:2.69-7.64) increased the recurrence risk. Moreover, portal vein invasion had a higher risk of death (HR=2.98, 95% CI: 1.73-4.98). Conclusion: There was significant association between AKAP12 DNA methylation and low risk of recurrence and long progression-free survival of hepatocellular carcinoma patients.

[The value of fractionated exhaled nitric oxide in the diagnosis of asthma-chronic obstructive pulmonary disease overlap syndrome].

Objective: To explore the diagnostic value of fractionated exhaled nitric oxide (FeNO) measurement in patients with asthma-chronic obstructive pulmonary disease(COPD) overlap syndrome (ACOS). Methods: Eighty-one patients with ACOS, 76 patients with asthma, 82 patients with COPD and 39 healthy non-smoking subjects were recruited in the study. Naku Lun breath analyzer was used to measure the level of FeNO in the 4 groups. Pulmonary function was also measured. The ROC curve was used to differentiate ACOS from patients with COPD. The correlation between FeNO and lung function was analyzed with Pearson correlation analysis. Results: The levels of FeNO in asthmatic group, COPD group, ACOS group and healthy group were (102.3±8.2)×10(9,) (23.7±0.6)×10(9,) (50.2±3.2)×10(9,) and (18.5±7.1)×10(9) respectively. Among the former 3 groups, the differences of FeNO were statistically significant (P<0.05). FeNO>29×10(9) was the best cutoff point to differentiate ACOS from COPD; the sensitivity was 80%, specificity was 73%, positive predictive value was 75%, and negative predictive value and accuracy was 79% and 77%. There was no correlation between FeNO and FEV(1)% or FEV(1)/FVC in ACOS, COPD and asthma groups (r=0.12, 0.11, P>0.05; r=0.11, 0.03, P>0.05; r=0.06, 0.08, P>0.05). Conclusion: FeNO is a good marker to help clinicians differentiate ACOS from COPD. FeNO>29×10(9) was the best cutoff point for the identification of patients with ACOS from COPD.

[Multiposition spiral suture of the lower uterine segment: a new technique to control the intraoperative bleeding of pernicious placenta previa].

Objective: To explore the efficacy and safety of multiposition spiral suture of the lower uterine segment, a new technique to control the intraoperative bleeding of pernicious placenta previa(PPP). Methods: From May 2014 to May 2015, 38 patients were diagnosed PPP in Tongji Hospital and cesarean sections were performed. After removing the placenta, multiposition spiral suture was used when massive bleeding occurred, and bilateral descending branches of uterine artery ligation was conducted when necessary. Results: 18 of the 38 PPP patients(47%,18/38)were diagnosed placenta accreta. The average cervical canal length of 38 PPP patients was(3.1±0.6)cm. There were 12 cases(32%, 12/38)with 4 regions sutured, 23 cases(61%, 23/38)with 2-3 regions sutured and 3 cases(8%, 3/38)with only posterior wall area sutured. Twelve cases(32%, 12/38)underwent uterine artery ligation, 3 cases(8%, 3/38)underwent uterine artery ligation and COOK balloon. None of them was postpartum hemorrhage or performing internal iliac artery embolization. Two patients received hysterectomy. The average blood loss in the operation was(1 696± 1 397)ml. In 16(42%,16/38)patients, the blood loss exceeded 1 500 ml, and the heaviest one was 4 500 ml. Three patients had haematuria in the first 3 days after the operation. No complication was found in 6 months after the operation. Conclusions: The multiposition spiral suture technique is a simple, safe and effective way to control the massive bleeding in the cesarean section of PPP patients. It is also beneficial for the recovery of the uterus.