[Clinical analysis of adverse reactions in patients with multidrug-resistant and rifampicin-resistant pulmonary tuberculosis treated with delamanid-containing regimen].

Objective: To explore the characteristics of adverse drug reactions during the 24-week therapy with delamanid-containing regimen for patients with multidrug-resistant and rifampicin-resistant pulmonary tuberculosis (MDR/RR-PTB). Methods: The prospective multicenter study was conducted from June 2020 to June 2023. A total of 608 eligible patients with MDR/RR-PTB were enrolled in 26 tuberculosis medical institutions in China including 364 males and 79 females, aged 39.6(19.0-68.0) years. Patients were treated with chemotherapy regimens containing delamanid. Patients were closely supervised during treatment of medication, and all adverse reactions occurring during treatment were monitored and recorded. The clinical characteristics of adverse reactions were evaluated by descriptive analysis. Chi-square test and multivariate logistic regression were used to analyze the related factors of QTcF interval prolongation (QT corrected with Fridericia's formula). Results: Of the 608 patients enrolled in this study, 325 patients (53.5%) reported 710 adverse events within 24 weeks of treatment. The top 6 most common complications were hematological abnormalities (143 patients, 23.5%), QT prolongation (114 patients, 18.8%), liver toxicity (85 patients, 14.0%), gastrointestinal reaction (41 patients, 6.7%), peripheral neuropathy (25 patients, 4.1%) and mental disorders (21 patients, 3.5%). The prolongation of QT interval mostly occurred in the 12th week after the first dose of medication. Serious adverse reactions occurred in 21 patients (3.5%). There were 7 patients (1.2%) with mental disorders, including 2 patients (0.3%) with severe mental disorders. Conclusions: The safety of dalamanid-based regimen in the staged treatment of MDR/RR-PTB patients was generally good, and the incidence of adverse reactions was similar to that reported in foreign studies. This study found that the incidence of QT interval prolongation in Chinese patients was higher than that reported overseas, suggesting that the monitoring of electrocardiogram should be strengthened when using drugs containing delamanid that may cause QT interval prolongation.

[Correlation between coronary artery tortuosity and poor prognosis in patients with septal hypertrophic cardiomyopathy].

Objective: To investigate the incidence of coronary artery tortuosity and its correlation with poor prognosis in patients with septal hypertrophic cardiomyopathy (HCM). Methods: This was a retrospective cohort study. Patients with septal HCM who were hospitalized in Fuwai Central China Cardiovascular Hospital and Zhengzhou University People's Hospital between December 1, 2017 and June 10, 2021 were selected. Non-HCM patients were matched by gender, age, and hypertension as control group. Septal HCM was divided into two groups based on the presence or absence of coronary artery tortuosity. Clinical baseline data and coronary angiography findings were compared using a multifactorial logistic analysis of the risk factors for coronary artery tortuosity. Patients were followed up until July 1, 2022, with the primary outcome being the composite endpoint of malignant arrhythmia, ischemic stroke and all-cause death. Incidence densities were compared between the coronary artery tortuosity and non-coronary artery tortuosity groups of septal HCM patients. The Cox risk-ratio model was used to analyze risk factors for primary outcomes in septal HCM patients. Results: There were 156 patients in the septal HCM group and 156 patients in the control group, both aged (57.0±11.4) years, and 75 (48.1%) were female. The incidence of coronary artery tortuosity was significantly higher in the septal HCM group than in the control group (63.5% vs. 36.5%, P<0.01), and the coronary artery tortuosity score was also higher in the septal HCM group than in the control group (P<0.01). Multiple logistic regression analysis showed that septal HCM was a risk factor for coronary artery tortuosity (OR=3.27, 95%CI: 2.02-5.29, P<0.01). In the septal HCM patients, after (2.5±1.2) years of follow-up, the incidence density of primary outcome was significantly higher in the coronary artery tortuosity group than in the non-coronary artery tortuosity group (P=0.02), while each on-point in coronary artery tortuosity score increased the risk of primary outcome by 53% for septal HCM patients (HR=1.53, 95%CI: 1.26-1.86, P<0.01). Conclusions: Patients with septal HCM are more prone to suffer coronary artery tortuosity and suffer from it to a greater extent. Coronary artery tortuosity is an important risk factor for adverse events in patients with septal HCM.

[Clinical features and prognosis of cutaneous melanoma].

Objective: To analyze the clinical features and prognosis of patients with cutaneous melanoma. Methods: The clinical data and follow-up data of 125 patients with cutaneous malignant melanoma (CMM) treated in the Tongji Hospital of Tongji Medical College of Huazhong University of Science and Technology between February 2008 and August 2019 were collected. Kaplan-Meier method and Log rank test were used for survival analysis, and Cox proportional risk regression model was used for impact factor analysis. Results: Among the 125 patients, 12 were stage Ⅰ, 62 were stage Ⅱ, 30 were stage Ⅲ, and 21 were stage Ⅳ; 76 were acral and 49 were non-acral. The median survival time was 44 months, and the 1-, 2-, and 5-year survival rates were 85.4%, 63.2% and 38.7%, respectively. Kaplan-Meier univariate survival analysis showed that Karnofsky performance status score, tumor stage, primary site, vascular infiltration, Ki-67, BRAF, lactate dehydrogenase (LDH), and surgical treatment were related to the prognosis of patients (P<0.05). The median overall survival (OS) time of patients receiving interferon treatment was 53 months, which was better than 40 months of patients not receiving interferon treatment, but the difference was not statistically significant (P=0.448). Among stage Ⅲ patients, the median OS time of patients receiving interferon therapy was 40 months, which was better than 17 months of patients not receiving interferon therapy (P=0.012). Among stage Ⅱ patients, the 1-, 2-, and 5-year survival rates of acral patients were 97.1%, 84.7%, and 65.8%, and the 1-, 2-, and 5-year survival rates of non-acral patients were 93.3%, 70.0% and 17.0%. The prognosis of patients with stage Ⅱ acral type was better than that of non-acral type (P=0.043). The median survival time of stage Ⅲ patients with acral type was 32 months, better than 17 months of non-acral type, but the difference was not statistical significance (P=0.164). The median survival time of acral type and non-acral type was 8 months and 11 months respectively (P=0.458). Cox multivariate analysis showed that tumor stage and preoperative LDH level were independent prognostic risk factors for cutaneous melanoma. Conclusions: Interferon treatment can improve the prognosis of patients with stage Ⅲ, and stage Ⅱ acral type patients have better prognosis than that of non-acral type patients. Tumor stage and preoperative LDH level were independent prognostic risk factors for cutaneous melanoma.

[Natural history and disease progression of chronic hepatitis B virus infection].

OBJECTIVE: To better understand and revise the natural history and disease progression of chronic hepatitis B virus (HBV) infection through analysis of a single-center large-scale cohort of indivi-duals with chronic HBV infection. METHODS: Patients with chronic HBV infection who had undergone liver biopsy in the Fifth Medical Center of Chinese People's Liberation Army (PLA) General Hospital from January 2014 to October 2020 were retrospectively recruited. Based on patient's hepatitis B e antigen (HBeAg) states and pathologic diagnosis, they were categorized into four disease progression statuses (or phases according to the old-terminology in the updated guidelines of chronic hepatitis B (CHB), such as European Association for the Study of the Liver (EASL) 2017, Clinical Practice Guidelines on the Management of Hepatitis B Virus Infection: HBeAg-positive chronic HBV infection (immune tolerance), HBeAg-positive CHB (immune active HBeAg positive), HBeAg-negative chronic HBV infection (inactive carrier), and HBeAg-negative CHB (immune reactive HBeAg negative). Then the demographic, laboratory tests and liver histological results of the patients in different disease progression stages were compared. Age differences between the two groups were evaluated using Mann-Whitney U test. RESULTS: A total of 760 eligible patients with a median age of 29 (interquartile range: 16-39) years were enrolled. Among them, 197 were underage individuals (age < 18 years) and 563 were adults; and 456 were males and 304 females. According to the pathological diagnosis, the patients were classified, and in each of the above four natural disease phases there were 173, 329, 95, and 163 individuals, respectively. Further comparison of the ages of the patients of the four disease progression statuses revealed that patients of HBeAg-negative CHB had a median age at 37 years, which was reasonably higher than those with HBeAg-positive CHB in immune active phase (37 vs. 24 years, P < 0.001), but was relatively younger than those with HBeAg-negative chronic HBV infection (37 vs. 39 years, P= 0.240). CONCLUSION: According to this study, it could be speculated that HBeAg-negative CHB patients probably not all reactivate from individuals of HBeAg-negative chronic HBV infection. Instead, certain HBeAg-negative CHB patients may also come from HBeAg-positive CHB patients who have undergone HBeAg clearance or seroconversion and still remain in the immune active state.

[Effect of histone deacetylase 2 and 4 activity on connective tissue disease associated pulmonary fibrosis in mice].

Objective: To investigate the effect of histone deacetylase (HDAC) activity on connective tissue diseases (CTD) associated pulmonary fibrosis (PF) in mice. Methods: A single tracheal administration of bleomycin induced PF in C57BL/6J male mice was performed to establish a PF model. The experimental mice were divided into three groups: bleomycin group (group B, n = 16) which was given bleomycin A2 physiological saline solution 2.5 µl/g body weight, saline group (Group C, n = 16) which was given physiological saline solution 2.5 µl/g body weight and no operation group (group N, n = 16). At 7, 14 and 21 days after administration, the animals were randomly killed and their specimens were collected. The activity of HDAC2 and HDAC4 was detected by colorimetry. Hematoxylin-eosin staining was used to evaluate pulmonary alveolitis and Masson staining for pulmonary fibrosis. The variance, correlation and binary variable correlation were analyzed. Results: The HDAC2 activity in lung tissue of mice in the bleomycin group was significantly higher than that in the no operation group (2.00±0.40 vs 1.00±0.23, P<0.05) and the saline group (2.00±0.40 vs 1.48±0.33, P<0.05). The HDAC2 activity in the bleomycin group was significantly higher than that in the no operation group (2.40±0.28 vs 1.00±0.23, P<0.01, 2.23±0.41 vs 1.00±0.23, P<0.01) and the saline group (2.40±0.28 vs 1.39±0.23, P<0.05, 2.23±0.41 vs 1.35±0.42, P<0.05). The change trend of HDAC2 activity between the bleomycin group and the saline group was different. There was no significant difference in HDAC4 activity in lung tissue of mice between the bleomycin group, the no operation group and the saline group. 14 days after tracheal administration, HDAC4 activity in the bleomycin group and the saline group were significantly higher than that in the no operation group (1.18±0.36 vs 1.00±0.12, P<0.01, 1.09±0.33 vs 1.00±0.12, P<0.01). HDAC2 activity in lung tissue of mice was positively correlated with pathological scores of alveolitis (r=0.428, P<0.01) and pulmonary fibrosis (r=0.508, P<0.01). HDAC4 activity in lung tissue of mice was positively correlated with the pathological scores of alveolitis (r=0.355, P<0.05) and pulmonary fibrosis (r=0.457, P<0.01). Binary linear regression analysis showed that HDAC2 activity had a stronger effect on the process of PF lesions than HDAC4 activity in lung tissue of mice. Conclusions: When pulmonary fibrosis occurred in mice, the activities of HDAC2 and 4 in pulmonary fibrosis were significantly increased. The activity of HDAC2 increased rapidly and lastingly, and the activity of HDAC4 fluctuated significantly and increased briefly. Changes in HDAC2 activity have stronger effects on alveolitis and fibrosis than HDAC4.

[Efficacy and safety comparison between pro-urokinase and reteplase in the treatment of patients with acute ST elevation myocardial infarction].

Objective: To compare the efficacy and safety of pro-urokinase and reteplase in the treatment of patients with acute ST elevation myocardial infarction (STEMI). Methods: STEMI patients, who received intravenous thrombolytic therapy in Henan STEMI registry between September 2016 and August 2018, were eligible for this study. A total of 5479 patients from 66 hospitals were screened and patients were divided into pro-urokinase group (n=638) and reteplase group (n=702) according to thrombolytic drugs. Data including patient demographics, risk factors, medical histories, patient information at admission, in-hospital treatment, time delays, and clinical events were collected. The clinical recanalization rate, in-hospital mortality, in-hospital death or treatment withdrawal, in-hospital main adverse cardiovascular and cerebrovascular events (MACCE, death or treatment withdrawal, congestive heart failure, reinfarction and ischemic stroke) and post-thrombolysis bleeding were compared between the two groups. Bleeding events were evaluated with Bleeding Academic Research Consortium (BARC) criteria. Results: The median age [61.8 (53.2, 69.0) vs. 62.6 (52.1, 69.8), P=0.833] or the proportion of women [23.0% (147/638) vs. 25.1% (176/702), P=0.385] were similar between the pro-urokinase and reteplase groups. Clinical recanalization rates were similar between the pro-urokinase and reteplase groups [82.1% (524/638) vs. 84.9% (596/702), P=0.172], and there was no difference in the median time from onset to thrombolysis [194.5 (135.0,290.0) min vs. 190 (126.0,292.0) min, P=0.431] and the median recanalization time [95 (67.5,120.0) min vs. 95 (71.0,119.0) min, P=0.561] between the two groups. There was no significant difference in in-hospital mortality [5.5% (35/638) vs. 5.1% (36/702), P =0.770], in-hospital all-cause mortality, treatment withdrawal [8.9% (57/638) vs.7.7% (54/702), P=0.410], and in-hospital MACCE [13.0% (83/638) vs. 10.4% (73/702), P=0.137] between pro-urokinase and reteplase groups. However, the incidence of post-thrombolysis bleeding was significantly higher in reteplase group than in pro-urokinase group [7.8% (55/702) vs. 3.8% (24/638), P=0.002]. Further analysis found that the incidence of oral bleeding and the BARC grades 1-2 bleeding were significantly higher in reteplase group than in pro-urokinase group, whereas the incidence of cerebral hemorrhage was similar between the two groups [0.6% (4/638) vs. 0.4% (3/702), P=0.715]. The comparison of efficacy and safety outcomes between the two groups after adjusting for baseline characteristics using general linear mixed models was consistent with those before the adjustment. There was no significant difference in in-hospital mortality, in-hospital death or treatment withdrawal, in-hospital MACCE after adjusting for baseline characteristics and post-thrombolysis bleeding between the two groups. Conclusions: Pro-urokinase and reteplase have similar clinical efficacy in the treatment of STEMI. In terms of safety, the incidence of cerebral hemorrhage is similar, while the incidence of BARC grades 1-2 bleeding and oral bleeding is higher in reteplase group than in pro-urokinase group, which has no impact on in-hospital outcomes.

Prognostic alternative splicing regulatory network of splicing events in acute myeloid leukemia patients based on SpliceSeq data from 136 cases.

This study aimed to create prognostic signatures to predict AML patients' survival using alternative splicing (AS) events. The AS data, RNA sequencing data, and the survival statistics of 136 AML patients were obtained from The Cancer Genome Atlas (TCGA) and TCGA SpliceSeq databases. Total 34,984 AS events generated from 8,656 genes, 2,583 of which were survival-associated AS events, were identified using univariate Cox regression. The prognostic models constructed using independent survival-associated AS events revealed that low-risk splicing better predicted patients' survival. ROC analysis indicated that the predictive efficacy of the alternate terminator model was best in the area under the curve at 0.781. Enrichment analysis revealed several important genes (TP53, BCL2, AURKB, PPP2R1B, FOS, and BIRC5) and pathways, such as the protein processing pathway in the endoplasmic reticulum, RNA transport pathway, and HTLV-I infection pathway. The splicing network of splicing events and factors revealed interesting interactions, such as the positive correlation between HNRNPH3 and CALHM2-13010-AT, which may indicate the potential splicing regulatory mechanism. Taken together, survival-associated splicing events and the prognostic signatures for predicting survival can help provide an overview of splicing in AML patients and facilitate clinical practice. The splicing regulatory network may improve the understanding of spliceosomes in AML.

[A novel electrochemiluminescence method for detecting insulin autoantibody].

Objective: To establish the electrochemiluminescence (ECL) method for insulin autoantibody (IAA) detection in China and preliminarily evaluate its application value. Methods: Proinsulin was labeled with Sulfo-tag and biotin, and then incubated with IAA in serum. The Meso Scale Discovery (MSD) streptavidin plate was used to capture the labeled antigen-antibody complex, which was subsequently detected by MSD electrochemiluminescence machine. IAA index ≥ 0.005 was used as a positive threshold. After optimizing the detection conditions, the correlation and consistency of ECL method and microplate radioimmunoassay were compared. Patients with type 1 diabetes mellitus (T1DM, n=55) and first-degree relatives (n=216) from the Department of Metabolism and Endocrinology of the Second Xiangya Hospital of Central South University from 2016 to 2018, and healthy volunteers (n=413) were selected as subjects, and their serum samples were tested to evaluate the clinical application value of ECL. Results: The optimized conditions included as follows: (1) The optimal concentration of biotin-proinsulin and Sulfo-tag-proinsulin was 800 ng/ml. (2) Serum acidification was achieved by incubating for 45 min. (3) Adding 35 µl diluted antigen, followed by adding 15 µl Tris-HCl to the reaction system, and then incubated with acidified serum overnight to obtain a higher signal-to-noise ratio (S/N). Twenty-two samples were repeatedly detected by ECL method at intervals of 3 months. There was no significant difference in IAA index (P=0.095) and the positive or negative results was completely consistent. The agreement rate between ECL method and microplate radioimmunoassay for IAA was 93.7% (119/127) (Kappa value: 0.858), and the index showed a significant positive correlation (correlation coefficient r=0.749, P<0.001). The area under the curve (AUC) of ECL assay was 0.764 (95%CI: 0.669 to 0.858). The positive rate of IAA in T1DM patients by ECL method was 52.73% (29/55), which was significantly higher than 0.76% (2/263) in healthy controls. The positive rate of IAA in T1DM first-degree relatives group was 0.93% (2/216), and there was no significant difference compared with healthy control group (P=0.854). Conclusion: ECL method of IAA has the features of high sensitivity and specificity, simple operation and no radioactive contamination, indicating its appreciable clinical application value.

Stepwise basis set selection.

The computational cost of quantum chemical methods grows rapidly with increasing level of theory and basis set size. At increasing costs, higher accuracies can be reached, forcing a compromise between cost and accuracy for most molecular systems. Heats of reaction, however, are mostly determined by a subset of atoms that experience significant bonding and/or electronic changes. To exploit this fact, the Stepwise Basis Builder (SBB) algorithm selectively adds basis functions to reactive atoms and maintains small basis sets on spectator atoms. This article introduces the SBB algorithm and how it chooses a basis for each atom, predicts calculation errors, and uses these predicted errors to reach target levels of accuracy. Benchmarks show SBB heats of reaction and activation barriers converge to values consistent with higher-quality calculations using a greatly reduced number of basis functions. © 2018 Wiley Periodicals, Inc.

Improvement in nitrogen fixation capacity could be part of the domestication process in soybean.

Biological nitrogen fixation (BNF) in soybeans is a complex process involving the interplay between the plant host and the symbiotic rhizobia. As nitrogen supply has a crucial role in growth and development, higher nitrogen fixation capacity would be important to achieve bigger plants and larger seeds, which were important selection criteria during plant domestication by humans. To test this hypothesis, we monitored the nitrogen fixation-related performance in 31 cultivated and 17 wild soybeans after inoculation with the slow-growing Bradyrhizobium diazoefficiens sp. nov. USDA110 and the fast-growing Sinorhizobium (Ensifer) fredii CCBAU45436. Our results showed that, in general, cultivated soybeans gave better performance in BNF. Electron microscopic studies indicated that there was an exceptionally high accumulation of poly-ß-hydroxybutyrate bodies in bacteroids in the nodules of all wild soybeans tested, suggesting that the C/N balance in wild soybeans may not be optimized for nitrogen fixation. Furthermore, we identified new quantitative trait loci (QTLs) for total ureides and total nodule fresh weight by employing a recombinant inbred population composed of descendants from a cross between a cultivated and a wild parent. Using nucleotide diversity (θπ), divergence index (Fst) and distribution of fixed single-nucleotide polymorphisms as parameters, we found that some regions in the total ureides QTL on chromosome 17 and the total nodule fresh weight QTL on chromosome 12 exhibited very low diversity among cultivated soybeans, suggesting that these were traits specially selected during the domestication and breeding process.

[Comparative analysis of the ankle joints in juvenile male soccer players with imaging].

OBJECTIVE: To investigate the characteristics of children male soccer players' ankle imaging features. METHODS: From October 2015 to February 2016, a total of 32 male children players of two soccer clubs in Guangzhou were enrolled in this study.The ages of all cases were from 10 to 14 years, average age was (11.3±0.9) years.A total of 15 male children in region ordinary primary and secondary school students were set as control group, ages were from 10 to 14 years, average age was (12.1±1.2) years. All objects' ankle were examined by X-ray for positive and lateral positions; routine CT scanning and then on the workstation restructuring for axial, coronal and sagittal slices; and examined by MR. MR scan was with special surface coil for ankle joint for horizontal axis T2WI; coronal T1WI; coronary proton density weighted imaging (PWI); sagittal T2WI with fat suppression; sagittal PWI with isotropic and fat suppression sequence of fast field echo. The ankle bone morphological structures were observed on X-ray; the ankle bone mineral density, cortical bone thickness and sesamoid bone quantity was being observed and measured on CT; and the tenosynovitis, Achilles tendinitis, synovitis, and cancellous bone edema signal were observed on MR. RESULTS: For study group, a total of 32 cases and 64 ankles joints were completed by X-ray, CT and MR examination.A total of 15 cases and 30 ankles joints were completed by X-ray in control group, 26 ankle joints were completed by CT scan and 22 ankle joints were completed by MR examination.X-ray examination showed there was no statistically significant difference between the two groups in ankle bone structure.CT showed that navicular bone CT value was (296±82) HU in research group and navicular bone CT value was (266±107) HU in control group, the difference was statistically significant (P=0.03). MR showed the incidence of diseases in research group that the tendon sheath peripheral inflammation was 92.2% (59/64), Achilles tendon lesions was 18.8%(12/64), edema of cancellous bone was 73.4% (47/64) , lateral malleous ligaments injuried was 43.8%(28/64), synovitis or effusion in posterior ankle was 87.5% (56/64). The incidence of diseases in control group was that tendon sheath peripheral inflammation was 31.8%(7/22), Achilles tendon lesions was 0/22, edema of cancellous bone was 0/22, lateral malleous ligaments injuried was 0/22, synovitis or effusion in posterior ankle was 54.5% (12/22). There was statistically significant difference between these two groups (all P<0.05). CONCLUSION: Compared with the control group, children's male soccer players ankle bone structure, bone cortex thickness and bone mineral density there were no obvious difference. In the tenosynovitis, Achilles tendinitis, synovitis, bone marrow edema, lateral malleous ligaments injuried were significantly higher than the control group.

[Efficacy of thrombolytic therapy using reteplase in cases with acute ST-segment elevation myocardial infarction: results from a multicenter clinical trial].

Objective: To evaluate the efficacy and safety of intravenous thrombolytic therapy using reteplase in patients with acute ST-segment elevation myocardial infarction (STEMI). Method: A total of 73 hospitals from Henan province took part in this clinical trials during October 2012 to October 2014, 1 226 cases (1 014 male (82.7%), mean age 59.0 (51.0, 66.0) years) with acute STEMI received reteplase as thrombolytic agent.Reperfusion rate was judged according to the clinical symptoms, electrocardiogram, myocardial enzymes and heart rhythm, and the rate of cardiovascular events and bleeding events during hospitalization was also observed.Bleeding events were evaluated with global utilization of streptokinase and tissues plasminogen activator for occluded coronary arteries (GUSTO) criteria.Subgroup analysis was performed to compare the effects of various thrombolysis timing (time from onset to thrombolysis≤6 h or 6-12 h) on reperfusion rate, cardiovascular events and bleeding events rate. Results: The reperfusion rate was 89.3% (1 089/1 219) at 120 minutes after the thrombolysis, average recanalization time was (59.96±26.86) minutes.The reperfusion rate of ≤6 h thrombolysis group was significantly higher than in 6-12 hours group (90.3% (988/1094) vs. 80.8% (101/125), P=0.001), while in-hospital mortality (2.6%(28/1 094) and 0.8% (1/125), P=0.352) and rate of bleeding (5.9%(64/1 094) and 5.6%(7/125), P=0.910) were similar between the two groups. The total in-hospital mortality after thrombolysis was 2.4% (29/1219), which was significantly higher in failed recanalization group than in recanalization group (10.8%(14/130) vs. 1.4%(15/1089), P< 0.001). The total rate of bleeding after thrombolysis was 5.8% (71/1219), there were 3 severe bleeding cases according to GUSTO classification (0.2%), all of them were cerebral hemorrhage, and 2 out of 3 cases died. Conclusions: Reteplase use is related to high recanalization rate and low cardiovascular events and bleeding rate and our results thus show that reteplase is a safe and effective thrombolytic agent for STEMI patients.

Marker-assisted selection in breeding silkworm strains with high tolerance to fluoride, scaleless wings, and high silk production.

In the silkworm (Bombyx mori), tolerance to fluoride and scaleless wings are controlled by the dominant gene Dtf (dominant tolerance to fluoride) and recessive gene nlw (no Lepidoptera wings), respectively, and these genes have been mapped by using simple sequence repeat and sequence tag site markers. Marker-assisted evaluation and selection of silkworms with fluoride tolerance and scaleless wings were used for predicting fluoride resistance and scaleless wings in backcrossed animals. A silkworm strain was bred using this method, and its economic characteristics were found to be similar to those of commercial silkworms. These methods will therefore be useful for silkworm breeding programs and in screening for two or more characteristics of interest for segregating populations.

Toll-like receptor 3 polymorphism is not associated with neovascular age-related macular degeneration and polypoidal choroidal vasculopathy in the Chinese.

Toll-like receptor 3 (TLR3) variants in mainland northern Chinese patients with polypoidal choroidal vasculopathy (PCV) and neovascular age-related macular degeneration (nAMD) were investigated. The complete genes of TLR3, including all exons and the promoter region, were assessed using direct sequencing technology of 284 unrelated mainland northern Chinese individuals: 96 nAMD patients, 92 PCV patients, and 96 controls. Six single nucleotide polymorphisms were identified: rs5743303, rs5743305, rs5743312, rs3775291, rs3775290, and rs6830345. The distribution of TLR3 genotypes for nAMD and PCV was not significantly different compared with normal controls. This study indicates that the TLR3 gene polymorphism is not associated with nAMD and PCV in northern Chinese patients.

Marker-assisted selection in breeding silkworm strains with high silk production and resistance to the densonucleosis virus.

In the silkworm (Bombyx mori), resistance to the Zhenjiang (China) strain of the densonucleosis virus (DNV-Z) is controlled by the recessive gene nsd-Z (non-susceptible to DNV-Z), which is linked to 7 simple-sequence repeat markers. Marker-assisted evaluation and selection of DNV-Z-resistant silkworms were used for predicting DNV-resistance in backcrossed animals. A silkworm race was bred using this method, and its economic characteristics were found to be similar to those of commercial silkworm races. These markers will therefore be useful for silkworm breeding programs and in screening for densonucleosis resistance in segregating populations.

Sequence variability in two mitochondrial DNA regions and internal transcribed spacer among three cestodes infecting animals and humans from China.

Sequence variability in two mitochondrial DNA (mtDNA) regions, namely cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 4 (nad4), and internal transcribed spacer (ITS) of rDNA among and within three cestodes, Spirometra erinaceieuropaei, Taenia multiceps and Taenia hydatigena, from different geographical origins in China was examined. A portion of the cox1 (pcox1), nad4 genes (pnad4) and the ITS (ITS1+5.8S rDNA+ITS2) were amplified separately from individual cestodes by polymerase chain reaction (PCR). Representative amplicons were subjected to sequencing in order to estimate sequence variability. While the intra-specific sequence variations within each of the tapeworm species were 0-0.7% for pcox1, 0-1.7% for pnad4 and 0.1-3.6% for ITS, the inter-specific sequence differences were significantly higher, being 12.1-17.6%, 18.7-26.2% and 31-75.5% for pcox1, pnad4 and ITS, respectively. Phylogenetic analyses based on the pcox1 sequence data revealed that T. multiceps and T. hydatigena were more closely related to the other members of the Taenia genus, and S. erinaceieuropaei was more closely related to the other members of the Spirometra genus. These findings demonstrated clearly the usefulness of mtDNA and rDNA sequences for population genetic studies of these cestodes of socio-economic importance.

[Effect of hydroxyapatite based agents on the bonding properties of universal adhesives].

Objective: To study the effect of hydroxyapatite (HA) based agents on the bonding properties of universal adhesive with different application modes, and to provide evidence for the use of adhesives after desensitization treatment. Methods: Sixty impacted third molars were extracted and selected (acquired from Department of Oral and Maxillofacial Surgery, College of Stomatology, Xi'an Jiaotong University). Four third molars were used to prepare 1 mm thick dentin disks and treated with 1% citric acid to simulate sensitive tooth models. The dentin surfaces were observed by scanning electron microscope (SEM) after treating with no desensitization (control group), desensitized by HA based toothpaste Biorepair and Dontodent Sensitive respectively (desensitizing toothpaste A group and B group), or HA paste treatment (desensitizing paste group ) (n=2 per group). The remaining teeth were selected to expose the mid-coronal dentin and establish dentin sensitivity models. Then, the specimens were divided into 4 former groups and received corresponding treatment. Each group was randomly divided into 2 subgroups, and intermediately strong universal adhesive (G-Premio Bond) was applied on the desensitized dentin by either etch-and-rinse mode or self-etch mode. Resin-dentin slice specimens (n=4 per subgroup), microtensile specimens (n=20 per subgroup) and slice specimens (n=6 per subgroup) were prepared. The microstructure and nanoleakage of the adhesive interfaces were observed by scanning electron microscopy (SEM). The microtensile strength (bond strength) and fracture mode were tested and recorded. The water permeability of the adhesive interface was observed by laser scanning confocal microscopy (LSCM). Results: SEM showed that desensitizing toothpaste and desensitizing paste could partially or entirely occlude most of the dentin tubules. For the etch-and-rinse mode, the bond strength of specimens treated with toothpaste A [(40.98±4.60) MPa], toothpaste B [(40.89±4.64) MPa] and HA paste [(41.48±3.65) MPa] was significantly higher than that of the control group [(38.58±4.28) MPa] (F=3.89,P<0.05). There was no statistically significant difference in bond strength among the 4 subgroups for self-etch modes (F=0.48,P>0.05). After desensitization, the bond strength of the control group and desensitizing groups in the self-etch mode was significantly higher than that in the etch-and-rinse mode (P<0.05). The overall fracture modes were mixed failure and interfacial failure in the control group and desensitizing groups. SEM showed speckled silver-stained particles deposited along the bottom of the hybrid layer on the bond interface of etch-and-rinse mode, and there were few silver-stained particles deposited on the bond interface of self-etch mode. LSCM showed continuous linear penetration in the hybrid layer of etch-and-rinse mode subgroups and discontinuous linear penetration in the hybrid layer of self-etch mode subgroups. Conclusions: HA based desensitizers have no adverse effect on the bond strength of intermediately strong universal adhesive and show good bonding performance accompanied with the self-etch mode.

Influence of RNAi knockdown for E-complex genes on the silkworm proleg development.

Larvae of many holometabolous insects possess abdominal appendages called prolegs. Lepidoptera larvae have prolegs in the segments A3-A6. Functions of Lepidoptera hox genes on these abdominal appendages development is still a controversial issue. In this article, we report the use of double strand RNA (dsRNA)-mediated interference (RNAi) to dissect the function of some hox genes, specifically E-complex genes Ubx, abd-A, and Abd-B, in the ventral appendage development of the Lepidoptera silkworm, Bombyx mori. We found that Ubx RNAi caused leg identity in A1 segment, abd-A RNAi caused severe defect of abdominal prolegs and Abd-B RNAi allowed proleg identity in more posterior abdominal segments. These results confirm that Lepidoptera hox genes Ubx and Abd-B have evolved the repressing function to ventral appendage development, which is similar to those of Drosophila. However, Lepidoptera abd-A might have been modified distinctively during evolution, and has important roles in directing the development of prolegs.

Effect of ICSI on gene expression and development of mouse preimplantation embryos.

BACKGROUND: In vitro culture (IVC) and IVF of preimplantation mouse embryos are associated with changes in gene expression. It is however not known whether ICSI has additional effects on the transcriptome of mouse blastocysts. METHODS: We compared gene expression and development of mouse blastocysts produced by ICSI and cultured in Whitten's medium (ICSI(WM)) or KSOM medium with amino acids (ICSI(KSOMaa)) with control blastocysts flushed out of the uterus on post coital Day 3.5 (in vivo). In addition, we compared gene expression in embryos generated by IVF or ICSI using WM. Global pattern of gene expression was assessed using the Affymetrix 430 2.0 chip. RESULTS: Blastocysts from ICSI fertilization have a reduction in the number of trophoblastic and inner cell mass cells compared with embryos generated in vivo. Approximately 1000 genes are differentially expressed between ICSI blastocyst and in vivo blastocysts; proliferation, apoptosis and morphogenetic pathways are the most common pathways altered after IVC. Unexpectedly, expression of only 41 genes was significantly different between embryo cultured in suboptimal conditions (WM) or optimal conditions (KSOM(aa)). CONCLUSIONS: Our results suggest that fertilization by ICSI may play a more important role in shaping the transcriptome of the developing mouse embryo than the culture media used.

Relative sorption coefficient: Key to tracing petroleum migration and other subsurface fluids.

The accumulation and spatial distribution of economically important petroleum in sedimentary basins are primarily controlled by its migration from source rocks through permeable carrier beds to reservoirs. Tracing petroleum migration entails the use of molecular indices established according to sorption capacities of polar molecules in migrating petroleum. However, little is known about molecular sorption capacities in natural migration systems, rendering these indices unreliable. Here, we present a new approach based on a novel concept of relative sorption coefficient for quantitatively assessing sorption capacities of polar molecules during natural petroleum migration. Using this approach, we discovered previously unrecognized "stripping" and "impeding" effects that significantly reduce the sorption capacities of polar compounds. These discoveries provide new insights into the behaviors of polar compounds and can easily explain why traditional molecular indices yield incorrect information about petroleum migration. In light of these new findings, we established new molecular indices for tracing petroleum migration. We demonstrate via case studies that the newly established indices, unlike traditional molecular indices, are reliable and effective in tracing petroleum migration. Our approach can be applied to diverse basins around the world to reveal distribution patterns of petroleum, which would decrease environmental risks of exploration by reducing unsuccessful wells.