[Susceptibility study on the germline rare variants of bromodomain and extraterminal domain protein family-encoding genes and patients with cancer living in some regions of China].

Objective: To explore the relationship between germline rare variants of bromodomain and extraterminal domain (BET) protein family-encoding genes and susceptibility to cancer in some regions of China. Methods: Capturing probes were designed for bromodomain-containing protein 2 (BRD2), BRD3 and BRD4 genes, and Illumina high-throughput sequencing platform was used to conduct targeted sequencing of genomic DNA of peripheral blood leukocytes from 1 673 patients with cancer and 1 661 individuals without cancer recruited between October 2015 and July 2018 from Chinese PLA General Hospital, the Second Affiliated Hospital of Guangxi Medical University, People's Hospital of Macheng City, Hubei Province and Geneplus-Beijing Co. Ltd. Mutation detection and analysis were carried out according to the genome analysis toolkit (GATK) best practice guidelines, ANNOVAR and VEP software were used for annotation, and germline rare variants in BET family were screened. To determine potential pathogenic germline rare variants, clinical and experimental evidence was obtained from the ClinVar database and SIFT and Polyphen-2 softwares were used to predict pathogenicity. Fisher's exact test was used to compare the difference of the carrying rate of variants in the case group and the control group, and multivariate regression analysis was performed with the SKAT software with sex and age used as covariates. Results: Among the 1 673 cancer patients, 911 were males and 762 were females, with the mean age was (57.9±11.7) years. There were 1, 111 cases (66.4%) of lung cancer, 266 cases (15.9%) of colorectal cancer, 186 cases of breast cancer (11.1%), and 110 cases (6.6%) of esophagus or gastric cancer. In the same period 1, 661 non-tumor control individuals were recruited, including 821 males and 840 females, with the mean age was (44.5±13.9) years. It was observed that there were 4 potential pathogenic germline rare variants in BRD2 gene carried by 17 patients with cancer, 5 potential pathogenic germline rare variants in BRD3 gene and 8 potential pathogenic germline rare variants in BRD4 gene. The carrying rate of potential pathogenic germline rare variants in BRD2 gene in cancer patients was 1.02% (17/1 673), significantly higher than that in controls without cancer [0 (0/1 661); OR=+∞, 95%CI: 4.81-+∞, P<0.001]. The carrying rate of potential pathogenic germline rare variants in BRD3 gene in cancer patients was 0.24% (4/1 673), and the difference was not statistically significant compared with controls without cancer [0.12% (2/1 661); OR=1.99, 95%CI: 0.46-10.47, P=0.690]. The carrying rate of potential pathogenic germline rare variants in BRD4 gene in cancer patients was 0.18% (3/1 673), and the difference was not statistically significant compared with controls without cancer [0.36% (6/1 661); OR=0.50, 95%CI: 0.14-2.08, P=0.340]. Furthermore, the dataset of whole exome sequencing of Chinese individuals in "Huabiao Project" was used as an additional control, and the rate of carrying BRD2 rare variants in cancer patients was 17/3 346 (0.51%), significantly higher than that in controls without cancer [0.07% (3/4 154); OR=7.07, 95%CI: 2.32-22.83, P<0.001]. Among the 17 patients carrying 4 potentially pathogenic germline rare variants of BRD2 gene, 9 were patients with lung cancer, 6 were patients with colorectal cancer, 1 was patient with breast cancer, and 1 was patients with esophagus or gastric cancer. The carrying rate of potential pathogenic germline rare variants in BRD2 gene in lung cancer patients was 0.81 (9/1 111), significantly higher than that in controls without cancer [0(0/1 661); OR=+∞, 95%CI: 3.95-+∞,P<0.001]. The carrying rate of potential pathogenic germline rare variants in BRD2 gene in patients with colorectal cancer was 2.26% (6/266), significantly higher than that in controls without cancer [0(0/1 661); OR=+∞, 95%CI: 9.03-+∞, P<0.001]. Wilcoxon rank-sum test results showed that patients with colorectal cancer carrying BRD2 rare variants had an earlier age at diagnosis [(47.0±7.4) vs (57.2±12.1) years old, P=0.017]. Conclusions: BRD2 gene may be served as a candidate genetic susceptibility gene for lung cancer and colorectal cancer. Carrying BRD2 potential pathogenic germline rare variants is associated with higher risk of lung cancer and colorectal cancer, and with earlier age of colorectal cancer.

Structural and functional comparisons of the environmental microbiota of pond and tank environments at different locations for the commercial aquaculture of American shad.

The environmental microbiota plays a significant role in the growth and development of aquatic life. In recent years, American shad has become an important economic aquaculture species in China. However, information on the correlation between the growth of American shad and the aquaculture environment is limited. Through 16S rDNA-based sequencing, the microbiota communities in ponds at different locations (Jiangyin and Yancheng in Jiangsu, China) were investigated. The results showed that the richness and diversity of the microbiota in the pond were greater than those in the tank at the same location. At the phylum level, Proteobacteria and Firmicutes were more abundant in the samples from Jiangyin than in those from Yancheng. In addition, the ratio of Firmicutes to Bacteroidetes was larger in the JYT samples than in the YCT samples, which implied that the American shad cultured in the Jiangyin environment tended to be fatter than those cultured in Yancheng. The body weight data also confirmed this finding. Moreover, the proportions of functional annotations in the samples from the Jiangyin and Yancheng environments were similar, but there were differences between the overall levels. Our results highlighted the correlations between the environmental microbiome and the growth tendency of American shad.

[Clinical analysis of 61 patients with thrombotic thrombocytopenic purpura].

OBJECTIVE: To analyze and summarize the clinical features, diagnosis, treatment and prognosis of 61 patients with thrombotic thrombocytopenic purpura (TTP), so as to improve the ability of diagnosis and treatment. METHODS: The clinical data of 61 TTP patients admitted to Peking University People's Hospital from January 2004 to March 2019 were retrospectively analyzed, and the clinical manifestations, blood routine, hemolysis indicators, and von Willebrand factor lyase (von Willebrand factor-cleaving protease, vWF-CP, also known as ADAMTS13) of these patients were observed. According to the outcome at the time of discharge, they were divided into survival group and death group, and the differences in clinical characteristics, neutrophil to lymphocyte ratio (NLR) and plasma exchange between the two groups were compared. The PLASMIC scores were calculated and compared with ADAMTS13 to determine the accuracy of the PLASMIC score in predicting ADAMTS13. RESULTS: Among the 61 TTP patients, 22 were males and 39 were females, with an average age of (48±17) years. In the study, 48 cases had pentalogy, only 9 had triad, and the remaining 4 had no neuropsychiatric symptoms. Twenty-seven cases (44.3%) died and 34 cases (55.7%) survived. Among the 61 TTP patients, the platelet count was (12.9±9.5)×109/L, the hemoglobin (66.5±20.7) g/L, the percentage of erythrocyte fragments 3% (2%, 7%), and the plasma free hemoglobin increased to 360 (200, 457) mg /L, and the lactate dehydrogenase 1 508 (811, 2 133.8) U/L. The blood clotting was basically normal. The ADAMTS13 value of 30 patients was 49.0 (40.8, 61.3) µg/L, the ADAMTS activity of 10 patients was < 5%, and the remaining 21 patients were not checked. The PLASMIC score was 6-7 in 58 cases, 5 in 2 cases, and 4 in 1 case. The PLASMIC score predicted the decreased activity or the reduction of ADAMTS with a sensitivity as high as 97.5%. The NLR in the death group was higher than that in the survival group, but the difference was not statistically significant (P>0.05). The total amount and frequency of plasma exchange (PEX) in the death group were significantly less than those in the survival group, and the difference was statistically significant (P < 0.05). There was no significant difference in the treatment of glucocorticoids and human immunoglobulin between the two groups (P>0.05). CONCLUSION: PEX can significantly improve the survival rate of TTP patients. PLASMIC score can easily and quickly predict the possibility of ADAMTS13 activity reduction, which is beneficial to the early diagnosis of TTP and PEX treatment. NLR can reflect the systemic inflammatory process, but its significance in TTP needs further study.

The activity and role of autophagy in the pathogenesis of diabetic nephropathy.

OBJECTIVE: Evidence suggested that deficiency of autophagy is involved in the pathogenesis of diabetic nephropathy (DN). However, some recent studies have also shown that autophagy is activated in renal cells under diabetic conditions. In this review, we discuss whether autophagy is inactivated in renal cells in DN as well as the therapeutic potential of autophagy for treating DN, in order to aid future investigation in this field. MATERIALS AND METHODS: Relevant information, original research articles and reviews, were gathered primarily through a search in PubMed and Cochrane database. The activity and role of autophagy, as well as the relevant signaling pathways, were analyzed in different intrinsic renal cells, including podocyte, renal tubular epithelial cell, glomerular mesangial and endothelial cells. RESULTS: The upstream of autophagic pathway, but not whole pathway, was predominately studied in these intrinsic renal cells, such as the induction of autophagy, an amount of autophagic vacuoles and so on. In most cases, autophagic inactivation occurred, which is an important mechanism underlying DN progression. Targeting the autophagic pathway to activate autophagy activity might have renoprotective effect. However, autophagic activation was also found in a few studies, in which there was a debate on the role of activated autophagy: mounting an adaptive response or leading to autophagic apoptosis. CONCLUSIONS: The downstream of autophagic pathway, including the degradation of autophagic vacuoles, and lysosomal function, should be well studied to clarify the activity and role of autophagy in the progression of DN. Autophagy activation is likely a potential therapy for combatting DN.

Larva-mediated chalkbrood resistance-associated single nucleotide polymorphism markers in the honey bee Apis mellifera.

Chalkbrood is a disease affecting honey bees that seriously impairs brood growth and productivity of diseased colonies. Although honey bees can develop chalkbrood resistance naturally, the details underlying the mechanisms of resistance are not fully understood, and no easy method is currently available for selecting and breeding resistant bees. Finding the genes involved in the development of resistance and identifying single nucleotide polymorphisms (SNPs) that can be used as molecular markers of resistance is therefore a high priority. We conducted genome resequencing to compare resistant (Res) and susceptible (Sus) larvae that were selected following in vitro chalkbrood inoculation. Twelve genomic libraries, including 14.4 Gb of sequence data, were analysed using SNP-finding algorithms. Unique SNPs derived from chromosomes 2 and 11 were analysed in this study. SNPs from resistant individuals were confirmed by PCR and Sanger sequencing using in vitro reared larvae and resistant colonies. We found strong support for an association between the C allele at SNP C2587245T and chalkbrood resistance. SNP C2587245T may be useful as a genetic marker for the selection of chalkbrood resistance and high royal jelly production honey bee lines, thereby helping to minimize the negative effects of chalkbrood on managed honey bees.

Quantum chemical elucidation of the mechanism for hydrogenation of TiO2 anatase crystals.

Hydrogenation of TiO2 is relevant to hydrogen storage and water splitting. We have carried out a detailed mechanistic study on TiO2 hydrogenation through H and∕or H2 diffusion from the surface into subsurface layers of anatase TiO2 (101) by periodic density functional theory calculations implementing on-site Coulomb interactions (DFT + U). Both H atoms and H2 molecules can migrate from the crystal surface into TiO2 near subsurface layer with 27.8 and 46.2 kcal∕mol energy barriers, respectively. The controlling step for the former process is the dissociative adsorption of H2 on the surface which requires 47.8 kcal∕mol of energy barrier. Both hydrogen incorporation processes are expected to be equally favorable. The barrier energy for H2 migration from the first layer of the subsurface Osub1 to the 2nd layer of the subsurface oxygen Osub2 requires only 6.6 kcal. The presence of H atoms on the surface and inside the subsurface layer tends to promote both H and H2 penetration into the subsurface layer by reducing their energy barriers, as well as to prevent the escape of the H2 from the cage by increasing its escaping barrier energy. The H2 molecule inside a cage can readily dissociate and form 2HO-species exothermically (ΔH = -31.0 kcal∕mol) with only 26.2 kcal∕mol barrier. The 2HO-species within the cage may further transform into H2O with a 22.0 kcal∕mol barrier and 19.3 kcal∕mol exothermicity relative to the caged H2 molecule. H2O formation following the breaking of Ti-O bonds within the cage may result in the formation of O-vacancies and surface disordering as observed experimentally under a high pressure and moderately high temperature condition. According to density of states analysis, the projected density of states of the interstitial H, H2, and H2O appear prominently within the TiO2 band gap; in addition, the former induces a shift of the band gap position notably towards the conduction band. The thermochemistry for formation of the most stable sub-surface species (2HO and H2O) has been predicted. These results satisfactorily account for the photo-catalytic activity enhancement observed experimentally by hydrogenation at high temperatures and high pressures.

Better detection of Ig heavy chain and TCRγ gene rearrangement in plasma cell-free DNA from patients with non-Hodgkin Lymphoma.

UNLABELLED: Plasma cell-free DNA is the soluble DNA and tumor-derived DNA in plasma which has the same mutation as the tumor cellular DNA. This study aimed at comparing the properties of plasma cell-free DNA with the biopsy's DNA in order to evaluate the clinical significance of IgH and TCRγ gene rearrangement in plasma cell-free DNA from patients with non-Hodgkin's Lymphoma. A total of 360 samples were studied. IgH (FR3A/VLJH) and TCRγ (TVG/TJX) were amplified by PCR. Results of plasma cell-free DNA were compared with biopsy's DNA and mononuclear cellular DNA respectively. Plasma cell-free DNA were successfully extracted from 288 cases of newly diagnosed, refractory and relapsed NHL in total 360 patients (80%).But nothing was found in the other 72 remittent patients. The positive percentage of IgH rearrangement in patients with B-NHL was 81% in plasma cell-free DNA and 77% in biopsy's DNA (P>0.05). As to the ratio of TCRγ rearrangement in patients with T-NHL, the former was 44%, and the latter was 39% (P>0.05). These results show tumor-derived DNA could be detected in tumor loaded plasma, even of underlying cancer patients. For NHL patients, detecting IgH and TCRγ gene rearrangement of plasma cell-free DNA has the same clinical significance as biopsy's DNA. Moreover, it's more simple, convenient and non-invasive. KEYWORDS: Lymphoma non-Hodgkin, plasma, cell-free DNA, gene rearrangement, immunoglobulin, heavy-chain gene, T-cell receptor.γ

Potentially inappropriate prescribing for insomnia in elderly outpatients in Taiwan.

OBJECTIVE: This study was based on Taiwan's National Health Insurance (NHI) claim records with the aim of identifying specific types of potentially inappropriate sedative-hypnotic prescribing in elderly outpatients with insomnia. The potentially inappropriate prescribing included duplicate treatment, excessive dosage and duration or treatment and prescribing of hypnotics that are too long-acting. MATERIAL AND METHODS: This cross-sectional study was based on annual outpatient claim data for 2001 released by Taiwan's Bureau of National Health Insurance (BNHI). A subset was created for patients aged 65 years or older and coded as having insomnia. Physician consultation claim data were extracted and merged in 1 claim file consisting of ICD-9-CM codes, patient demographic data, specialty of physicians, medical institution code and pharmaceutical prescription content. RESULTS: Elderly patients with insomnia constituted 216,994 of the 1,000,193 files surveyed on outpatient claims in 2001. The mean age was 74.33 years and gender distribution was nearly equal. Of the prescribed sleep medications, 41.26% were hypnotic benzodiazepines, 29.36% were hypnotic non-benzodiazepines and 29.38% were sedative-anxiolytics. Approximately 1 in 25 patients (4.12%) of all the patients prescribed hypnotics received duplicate treatment, 1 in 8 an inappropriately (12.27%) high dosage (daily dose > 1.5 DDD, Defined Daily Dose), 1 in 3 (32.25%) more than 28 DDD per prescription and 1 in 6 (17.52%) a drug with an effect which was too prolonged. Physicians tended to consider patient gender but not age when prescribing. Clear trends were found between the specialty of the physician and the type of inappropriate prescribing. The type of medical institution was significantly related only to the excessive quantity of medication prescribed. CONCLUSIONS: Elderly people in Taiwan with insomnia receive potentially inappropriate prescriptions for sleep medications. Similar data could possibly be extracted from similar databases in other countries throughout the world. Some of these potentially inappropriate prescriptions are avoidable in terms of restricting the length of outpatient sedative-hypnotic treatment, introducing hypnotics in small dosage forms and continuously educating clinicians on the safety of geriatric medication.

Washing and cryoprotectant effects on frozen storage of spent hen surimi.

Mince of 98-wk-old spent hens was washed two times with 0.1% NaCl. Portions of unwashed and washed mince were mixed with a cryoprotectant (CP) composed of 4% sucrose, 4% sorbitol, and 0.2% Na-tripolyphosphate and were immediately frozen and stored at -20 C. Mince without CP was run as control. Textural properties of the stored mince and surimi were measured at 1-mo intervals for 6 mo, after being thawed at 4 C overnight, ground with 3% NaCl, and heated at 90 C for 15 min. For freeze-thaw stability study, minces were subjected to six freeze-thaw cycles. Each freeze-thaw cycle was carried out at 1 mo of storage. Textural quality parameters (gel strength, breaking strength, deformation, protein solubility, expressible moisture, cooking yield, folding test, drip-loss, and sensory scores) were decreased in both unwashed and washed mince, mostly during the early stages of storage. Washed mince showed significantly better textural properties than unwashed mince. Washing protected the gel quality of the hen mince from degradation during frozen storage. Cryoprotectants could not protect the gel strength or breaking strength, but deformation was slightly improved. Water-retention properties were protected, and folding test and sensory scores were well preserved in the mince with added CP. Cryoprotectants had a beneficial effect on frozen, stored spent hen surimi to protect the elasticity and cohesiveness of the gel.

Differences in molar absorptivity of 4-NP with the reaction solution and apparatus affect ALP measurement.

We examined the differences in molar absorptivity of 4-NP obtained using different kits for ALP measurement and different instruments. The apparent molar absorptivity of 4-NP in the same reaction solution determined by six different instruments was 15.98, 16.72, 16.06, 17.00, 16.27, 17.62 and that using four different reaction solution kits for ALP with the same instrument was 16.90, 17.38, 17.72, 16.11. We measured ALP in three serum samples with six instruments using the same kit and in twelve serum samples with the same instrument using four kits. ALP activities measured using the same molar absorptivity value differed with the instrument(p < 0.01). However, those measured using the apparent molar absorptivity value for each instrument revealed no significant differences(p > 0.05). In conclusion, we suggest that standard material should be contained in each kit for enzyme measurement and the apparent epsilon for each kit and instrument should be obtained to minimize the systematic error caused by using the same epsilon in different laboratories.

Adulteration by synthetic therapeutic substances of traditional Chinese medicines in Taiwan.

The adulteration by synthetic therapeutic substances of traditional Chinese medicines has been reported on various occasions and has been a public health concern in Taiwan over the past several years. A large-scale effort was initiated in 1992 to screen traditional Chinese medicines that were suspected of adulteration with synthetic therapeutic substances. The term "adulteration" refers to traditional Chinese medicines that are tested and found to contain chemical substances not prescribed or labeled as part of the intended use. A total of 2,609 samples were collected by eight major general hospitals in Taiwan. Samples were collected through physicians' referrals during patients visits. The samples were analyzed by hospital pharmacists following the established standard procedures in comparison to references by thin-layer chromatography. An average of 23.7% (n = 618) of the samples collected from the eight hospitals were adulterated. Four samples with either a rheumatoid or an antiinflammatory indication contained six different kinds of adulterants. More than half (52.8%) of the adulterated traditional Chinese medicines contained two or more adulterants. The sources of adulterated samples and their claimed indications, as well as the most frequently detected synthetic therapeutic substances, are presented in this report. The controversies regarding the combination of synthetic therapeutic substances and traditional Chinese medicines without adequate labeling should be resolved through regulatory actions for better safety of drug use.

Case-control study of the effectiveness of different types of helmets for the prevention of head injuries among motorcycle riders in Taipei, Taiwan.

A total of 1,351 victims of motorcycle accidents, brought to one of 15 hospitals responsible for emergency care in Taipei, Taiwan, between August 1 and October 15, 1990, were enrolled in a case-control study to investigate the effectiveness of different types of helmets for the prevention of head injuries. A total of 562 of those with head injuries were assigned to the case group, while the remaining 789 victims without head injuries were considered as emergency room controls. The case group was subdivided into daytime and evening cases, according to the time of accident. For each daytime case, we took four pictures of passing motorcycles at the same time and place during the week after each accident. Of the 254 daytime cases, we successfully took pictures for 224 (88%) and identified 1,094 motorcycle riders in the pictures as street controls. Logistic regression analyses were used to determine the roles of the following variables in predicting risk of head injury: age, sex, riding position, weather, place of accident, helmet type, and motorcycle type, and status of helmet wearing. The relative risk of head injury among motorcycle riders was significantly reduced by wearing a full-face helmet, but not by wearing a full- or a partial-coverage helmet.

[Chemotherapy of biliary tract infections (XXIV). Biliary excretion and gallbladder tissue levels of mezlocillin].

Mezlocillin (MZPC), a new broad spectrum penicillin, was studied for its biliary excretion and levels in gallbladder tissues. The mean serum concentration immediately after intravenous administration of 2 g of MZPC in 6 cholecystectomized patients was found to be 535.5 +/- 43.5 micrograms/ml, while its concentration in their gallbladder bile was 1,500-6,500 micrograms/ml at 30 minutes-1.5 hours and its concentration in their common duct bile was 6,500 and 4,000 micrograms/ml at 2 hours and 2.5 hours, respectively. Besides, its gallbladder tissue level around 2 hours was 37.5-500 micrograms/g, averaging as high as 215.6 +/- 79.5 micrograms/g. Crossover method was made to compare the biliary excretion of MZPC and PIPC in 4 patients inserted with a T-tube in their common bile duct. Each patient received 2 g each of 2 antibiotics intravenously. In case 1 with normal liver function, both drugs revealed the peak concentration of 3,500 micrograms/ml or more. In case 2, the peak concentration was 3,625 micrograms/ml for MZPC but 2,125 micrograms/ml for PIPC. In case 3 with slight hepatic dysfunction, the peak concentration for MZPC and PIPC was 906.4 micrograms/ml and 375.2 micrograms/ml, respectively. In case 4 with moderate hepatic dysfunction, the peak concentration for MZPC and PIPC was 27.6 micrograms/ml and 14.7 micrograms/ml, respectively. Simultaneous intravenous administration of MZPC and PIPC was made to compare the biliary excretion of these 2 antibiotics in 1 patient with T-tube drain in common bile duct. The bile concentration was measured by HPLC method which was developed by our laboratory.(ABSTRACT TRUNCATED AT 250 WORDS)