[Evaluation of the emergency response strategies and measures on the epidemic of COVID-19 in Shenzhen, China].

Objectives: This study aimed to evaluate the effect of the strategies on COVID-19 outbreak control in Shenzhen, and to clarify the feasibility of these strategies in metropolitans that have high population density and strong mobility. Methods: The epidemic feature of COVID-19 was described by different phases and was used to observe the effectiveness of intervention. Hierarchical spot map was drawn to clarify the distribution and transmission risk of infection sources at different time points. The Susceptible-Exposed-Infectious-Asymptomatic-Recovered model was established to estimate case numbers without intervention and compare with the actual number of cases to determine the effect of intervention. The positive rate of the nucleic acid test was used to reflect the risk of human exposure. A survey on COVID-19 related knowledge, attitude and behaviors were used to estimate the abilities of personal protection and emergency response. Results: The epidemic of COVID-19 in Shenzhen experienced the rising, plateau and decline stage. The case number increased rapidly at the beginning, with short duration of peak period. Although the epidemic curve showed human-to-human transmission, the "trailing" was not obvious. From the spot map, during the intervention period, the source of infection was widely distributed. More cases and higher transmission risk were observed in areas with higher population density. After the effective intervention measures, both infection sources and the risk of transmission decreased. After compared with the estimated case numbers without intervention, actual number proved the COVID-19 control strategies were effective. The positive rate of nucleic acid test for high risk populations decreased and no new cases reported since February 16. Shenzhen citizens had high knowledge, attitude and behavior level, and high protection ability and emergency response. Conclusions: Although the response initiated by the health administration department played a key role at the early stage of the epidemic, it was not enough to contain the outbreak of COVID-19. The first-level emergency response initiated by provincial and municipal government was effective and ensured the start of work resumption after the Spring Festival. Metropolitans like Shenzhen can also achieve the goals of strategies and measures for containment and mitigation of COVID-19.

[A multicenter study of reference intervals for 15 laboratory parameters in Chinese children].

Objective: To establish comprehensive laboratory reference intervals for Chinese children. Methods: This was a cross-sectional multicenter study. From June 2013 to December 2014, eligible healthy children aged from 6-month to 17-year were enrolled from 20 medical centers with informed consent. They were assessed by physical examination, questionnaire survey and abdominal ultrasound for eligibility. Fasting blood samples were collected and delivered to central laboratory. Measurements of 15 clinical laboratory parameters were performed, including estradiol (E2), testosterone(T), luteinizing hormone(LH), follicle-stimulating hormone(FSH), alanine transaminase(ALT), serum creatinine(Scr), cystatin C, immunoglobulin A(IgA), immunoglobulin G(IgG), immunoglobulin M(IgM), complement (C3, C4), alkaline phosphatase(ALP), uric acid(UA) and creatine kinase(CK). Reference intervals were established according to central 95% confidence intervals for reference population, stratified by age and sex. Results: In total, 2 259 children were enrolled. Finally, 1 648 children were eligible for this study, including 830 boys and 818 girls, at a mean age of 7.4 years. Age- and sex- specific reference intervals have been established for the parameters. Reference intervals of sex hormones increased gradually with age. Concentrations of ALT, cystatin C, ALP and CK were higher in children under 2 years old. Serum levels of sex hormones, creatinine, immunoglobin, CK, ALP and urea increased rapidly in adolescence, with significant sex difference. In addition, reference intervals were variable depending on assay methods. Concentrations of ALT detected by reagents with pyridoxal 5'-phosphate(PLP) were higher than those detected by reagents without PLP. Compared with enzymatic method, Jaffe assay always got higher results of serum creatinine, especially in children younger than 9 years old. Conclusion: This study established age- and sex- specific reference intervals, for 15 clinical laboratory parameters based on defined healthy children.

Dl-3-n-butylphthalide protects the blood brain barrier of cerebral infarction by activating the Nrf-2/HO-1 signaling pathway in mice.

OBJECTIVE: The aim of this study was to explore whether Dl-3-n-butylphthalide (DBT) could protect blood-brain barrier (BBB) of mice with experimental cerebral infarction and the relevant mechanism. MATERIALS AND METHODS: Adult male CD-1 mice were selected as the study objects. The permanent middle cerebral artery occlusion (MCAO) model was prepared by Longa's modified suture-occluded method. The mice were randomly divided into 3 groups: the sham operation group (Sham group), the cerebral infarction model group (CI group) and the DBT (120 mg/kg) intervention group (DBT group). Neurologic function deficits were evaluated by Longa's modified scoring method after 24 h of permanent MCAO. The wet and dry weight method was used for measuring water content in brain tissues. 2% 2,3,5-triphenyltetrazolium chloride (TTC) staining method was applied to determine the volume of cerebral infarction. Changes in the protein and messenger ribonucleic acid (mRNA) expression levels of matrix metallopeptidase 9 (MMP-9), claudin-5, vascular endothelial growth factor (VEGF), glial fibrillary acidic protein (GFAP), NF-E2 related factor 2 (Nrf-2) and heme oxygenase 1 (HO-1) in ischemic brain tissues were detected using immunohistochemistry, Western blotting and quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). Ultrastructure changes in BBBs were observed under an electron microscope. RESULTS: DBT improved the neurologic function deficits of mice and reduced the infarction volume of mice with cerebral infarction. DBT alleviated edema and decreased the permeability of BBBs of mice with cerebral infarction. DBT down-regulated the expression of MMP-9 and up-regulated the expression of claudin-5 in brain tissues of mice with cerebral infarction. DBT increased the expressions of VEGF and GFAP. DBT improved the ultrastructure in capillary endothelial cells of BBBs and increased the expressions of Nrf-2 and HO-1. CONCLUSIONS: DBT may protect BBB by activating the Nrf-2/HO-1 signaling pathway, thus achieving its protective effect on the brain.

[Survey of economic burden of hepatitis B-related diseases in 12 areas in China].

Objective: Less surveys on the economic burden of hepatitis B (HB)-related diseases have been conducted in China, so the socioeconomic harm caused by the diseases is not clear and the key parameters for economic evaluation of hepatitis B prevention and treatment are lacking. This study aimed to analyze the direct, indirect and intangible expenditures of hospitalized patients with HB-related diseases during hospitalization and during a year in different areas of China. Methods: The hospitals for infectious diseases and the large general hospitals in 12 areas in China were selected in the study. All the inpatients with HB-related diseases were surveyed by cluster sampling of consecutive cases. The direct expenditure included direct medical cost and direct non-medical cost. The indirect expenditure, including work loss of patients and caregivers, were calculated by using human capital method for urban and rural populations in 12 areas. The intangible expenditure were reflected by willing to pay and stochastic tournament. The influencing factors of direct and indirect costs were identified by stepwise linear multi-variation regression analysis. Results: A total of 27 hospitals in 12 areas were included in the survey. A total of 4 718 cases were surveyed, the overall response rate was 77.7%. The average hospital stay was 29.2 days (27-34) and the hospitalization expenditure was averagely 16 832.80 yuan (RMB) per case, in which the highest proportion (61.2%) was medicine fees [10 365.10 yuan (RMB)]. The average direct expenditure and indirect expenditure were consistent with the severity of illness, which were 18 336.10 yuan (RMB) and 4 759.60 yuan (RMB) respectively, with the ratio of 3.85 ∶ 1. The direct medical expenditure [17 434.70 yuan (RMB)] were substantially higher than the direct non-medical expenditure [901.40 yuan (RMB)]. It was found that the hospitalization expenses was highest in direct medical expenditure and the transportation expenses was highest in direct non-medical expenditures. Among the average indirect expenditure, the loss of income for the patients [3 832.50 yuan (RMB)] was higher than that for the caregivers [927.20 yuan (RMB)]. The total direct and indirect expenditure was highest for liver transplantation, followed by severe hepatitis, hepatocellular carcinoma and decompensated cirrhosis, acute hepatitis B, compensated cirrhosis and chronic hepatitis B. The influencing factors for both direct and indirect expenditure were high hospital level, severity of hepatitis B, living in urban area, antiviral therapy, long hospitalization and monthly income of family. For average 3.74 outpatient visits and 1.51 hospitalization, the average annual direct, indirect and intangible expenditure for HB-related diseases were 30 135.30, 6 253.80 and 44 729.90 yuan (RMB) [totally 81 119.00 yuan (RMB)], accounting for 37.3%, 7.7% and 55.0%, respectively. Of the annual direct medical expenditure [28 402.80 yuan (RMB)], which were much higher than non-medical expenditure [1 732.50 yuan (RMB)], hospitalization expenditure [26 074.20 yuan (RMB)] was higher than outpatient visit expenditure [4 061.10 yuan (RMB)]. The annual indirect expenditures for outpatient visit and hospitalization were 763.60 and 5 490.10 yuan (RMB), respectively. Of the annual intangible expenditure, the highest was that for primary hepatocellular carcinoma, followed by cirrhosis, chronic hepatitis B, severe hepatitis B, liver transplantation and acute hepatitis B. Conclusions: A heavy economic burden has been caused by HB-related diseases in China, and patients are more likely to rely on medical service rather than non-medical service. It is necessary to take effective treatment measures to prevent the adverse outcome of HB related diseases and achieve significant economic benefits. The influence of HB related diseases on mental health of the people can be reflected by an economics term, intangible expenditure.

Effects of intermittent hypobaric hypoxia preconditioning on the expression of neuroglobin and Bcl-2 in the rat hippocampal CA1 area following ischemia-reperfusion.

This study was aimed at understanding the effect of intermittent hypobaric hypoxia preconditioning (IHHP) on neuroglobin (NGB) and Bcl-2 expression in the hippocampal CA1 region of rats following global cerebral ischemia-reperfusion. Wistar rats were randomly divided into sham, IHHP control, global cerebral ischemia-reperfusion (IR group), and IHHP+IR groups. The four-vessel occlusion rat model of Pulsinelli was used for the IR groups, in which the common carotid artery was occluded for 8 min before reperfusion. Thionin and immunohistochemical staining were used to observe NGB and Bcl-2 expression in the hippocampal CA1 region. Data was analyzed using the SPSS software. There was a significant increase in the number of surviving cells in the hippocampal CA1 region of the IHHP+IR group (119.5 ± 14) compared to the IR group (41.7 ± 3.8) (P < 0.05). There was a significant increase in the expression of NGB and Bcl-2 in the hippocampal CA1 region of the IHHP+IR group compared to the IR group. By upregulating hippocampal NGB and Bcl-2 expression, IHHP may play a role in neural protection by reducing hippocampal neuronal apoptosis following IR.

Isolation and partial characterization of novel genes encoding acidic cellulases from metagenomes of buffalo rumens.

AIMS: To clone and characterize genes encoding novel cellulases from metagenomes of buffalo rumens. METHODS AND RESULTS: A ruminal metagenomic library was constructed and functionally screened for cellulase activities and 61 independent clones expressing cellulase activities were isolated. Subcloning and sequencing of 13 positive clones expressing endoglucanase and MUCase activities identified 14 cellulase genes. Two clones carried two gene clusters that may be involved in the degradation of polysaccharide nutrients. Thirteen recombinant cellulases were partially characterized. They showed diverse optimal pH from 4 to 7. Seven cellulases were most active under acidic conditions with optimal pH of 5.5 or lower. Furthermore, one novel cellulase gene, C67-1, was overexpressed in Escherichia coli, and the purified recombinant enzyme showed optimal activity at pH 4.5 and stability in a broad pH range from pH 3.5 to 10.5. Its enzyme activity was stimulated by dl-dithiothreitol. CONCLUSIONS: The cellulases cloned in this work may play important roles in the degradation of celluloses in the variable and low pH environment in buffalo rumen. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provided evidence for the diversity and function of cellulases in the rumen. The cloned cellulases may at one point of time offer potential industrial applications.

Finite element analysis of four thread-form configurations in a stepped screw implant.

The purpose of this study was to determine the optimal thread form configuration for an experimental stepped screw implant. Two-dimensional finite element analysis was applied to model the experimental stepped screw implant in a standard cross-section of the posterior human mandible digitized from a CT-generated patient data set. Four different thread form configurations: v-thread (V), thin-thread (T), and two square-thread forms of 0.24 mm (S1) and 0.36 mm (S2) thread width were compared under oblique load in normal cortical bone condition. The support-type constraint position changed from middle to the base of the bone segment. In middle support-type constraint position only the thin-thread (T) model demonstrated significantly different stress distribution from the other three models, however, in base support-type constraint position T and S1 models demonstrated significantly different stress distribution from the other two models. The results implies that v-thread (V) or large square-thread (S2) are optimal thread form for the experimental stepped screw implant. While, minimal support constraints allow clearer differentiation of the stress picture between the different stepped screw types at the trabecular bone-implant interface.

Cloning and characterization of the rpfC gene of Xanthomonas oryzae pv. oryzae: involvement in exopolysaccharide production and virulence to rice.

rpfC is one of a cluster of genes which coordinately regulate the synthesis of extracellular enzymes and exopolysaccharide and pathogenicity in Xanthomonas campestris pv. campestris, the black rot pathogen of brassicas. An rpfC homolog which could functionally complement an rpfC mutant of X. campestris pv. campestris was identified in Xanthomonas oryzae pv. oryzae and the gene was characterized. Mutation of this gene in X. oryzae pv. oryzae had no effect on extracellular enzymes, but exopolysaccharide synthesis and virulence to rice were substantially reduced.

Identification of a nodD-like gene in Frankia by direct complementation of a Rhizobium nodD-mutant.

Clones from a Frankia At4 gene bank were pooled into groups and mass conjugated into a nodD mutant of Rhizobium leguminosarum bv. viciae by triparental matings. When peas were inoculated with the pooled transconjugants, nodulation was observed. A plasmid, pAt2GX containing Frankia DNA, was isolated from bacteria recovered from these nodules. This plasmid was shown to complement a nodD mutant of R. leguminosarum bv. viciae. Thus pAt2GX contains a Frankia gene that is functionally equivalent to nodD of R. leguminosarum bv. viciae.

[Construction of Frankia genomic libraries and isolation of clones homologous to nodulation genes from Rhizobium leguminosarum].

High molecular genomic DNAs were isolated by using the lysozyme plus achromopeptidase system from Frankia strains At4, Ccol and Hr16, the root nodule endophytes of Alnus, Casuarina and Hippophae respectively, and used to construct genomic libraries in pLAFR1, a broad host range cosmid vector within many gram-negative hosts. The genomic libraries were screened by in situ colony hybridization to identify clones homologous to common nodulation genes of Rhizobium leguminosarum, based on the sequence homology of EcoRI-digested Frankia total DNA to nodABC from Rhizobium meliloti. Several clones showing relatively strong hybridization were found, the recombinant plasmid was isolated, and their homology with Rhizobium nodulation genes was confirmed by spot hybridization. Further work on these positive clones is now underway.

Identification and DNA sequence of fixZ, a nifB-like gene from Rhizobium leguminosarum.

Previously, several mutants which nodulated peas but which failed to fix nitrogen were isolated following Tn5 mutagenesis of pRL 1JI, a symbiotic plasmid of Rhizobium leguminosarum. Two of these alleles, fix52::Tn5 and fix137::Tn5 were in a region of pRL 1JI which hybridized to a probe that contained the nifA gene and the amino-terminal region of the nifB gene of Klebsiella pneumoniae. The nitrogen fixation defect of the fix52::Tn5 mutant strain was corrected by a 2.0kb fragment of the corresponding wild-type DNA cloned in a wide host-range plasmid. The DNA sequence of this region revealed an open reading frame corresponding to the gene within which the fix52::Tn5 allele was located. The polypeptide corresponding to this open reading frame had a deduced molecular weight of 39,936 and the gene was termed fixZ. The deduced amino acid sequence of the fixZ gene product contained two clusters of cysteine residues, suggesting that the protein may contain an iron-sulphur cluster. The sequence of the fixZ polypeptide was very similar to the sequence of the K. pneumoniae nifB gene (provided by W. Arnold and A. Pühler) which is required for the synthesis of the FeMo-cofactor of nitrogenase. It was shown that the previously observed hybridization was due to homology between the amino terminal regions of fixZ and nifB. Upstream from fixZ was found another open reading frame whose 5' terminus was not established, but within which was located the fix137::Tn5 allele. This gene was termed fixY. The deduced amino acid sequence of the sequenced part of fixY showed similarity to that of the regulatory nifA gene of K. pneumoniae (provided by W. J. Buikema and F. M. Ausubel). Thus in R. leguminoarum the fix genes that correspond to the nifA and nifB genes are in the same relative orientation as in K. pneumoniae.

Cloning of the symbiotic region of Rhizobium leguminosarum: the nodulation genes are between the nitrogenase genes and a nifA-like gene.

The region of the Rhizobium leguminosarum plasmid pRL1JI involved in nodulation and nitrogen fixation has been cloned on a series of four overlapping cosmid clones. These clones represent 60 kb of pRL1JI DNA on which a series of Tn5-induced fix and nod alleles have been identified, with the two most distant alleles being separated by 45 kb of DNA. The mutant alleles fell into three groups, two clusters of fix alleles separated by one cluster of nod alleles. Within one group of fix alleles, DNA homologous to the nifA gene of Klebsiella pneumoniae has been identified, whereas the pRL1JI DNA homologous to the K. pneumoniae nitrogenase genes is present within the other group of fix alleles.