[Correction model of the sampling time error on the blood trough concentration of tacrolimus in non-sustained-release dosage form for renal transplant recipients].

Objective: To establish correction model of the sampling time error on the blood trough concentration of tacrolimus in non-sustained-release dosage form for renal transplant recipient and improve the accuracy of drug dose assessment and clinical adjustment in renal transplant recipients. Methods: Visit records of 206 outpatients in the Department of Transplantation, Nanfang Hospital, Southern Medical University were retrospectively collected from October 15, 2022 to October 30, 2022. The distribution of sampling time of tacrolimus blood drug concentration was described and the time range of correction was determined. Twenty inpatients after renal transplantation in the Department of Transplantation, Nanfang Hospital, Southern Medical University from October 1, 2022 to November 30, 2022 were prospectively included, and their demography data, laboratory test results during follow-ups, and CYP3A5 genotype were collected. The patients took tacrolimus in non-sustained-release dosage form every 12 h starting from 19∶30 on the day of admission. Peripheral blood samples were collected from the patients on the second day of admission at 7∶30 and on the third day at 6∶00-10∶00 every 30 minutes to test the blood concentration of tacrolimus. Using the collection time as the independent variable and the blood tacrolimus concentration as the dependent variable, a simple linear regression was performed to fitting a linear model of tacrolimus blood concentration-sampling time. Multiple linear regression was performed to analyze the influencing factors of the tacrolimus metabolic rate within a specific period and generate the regression equation. Results: The 206 outpatients aged (46±13) years, including 131 males (63.6%). The time gap [M (Q1, Q3)] between the sampling time of the follow-up outpatients and standard C12 was 24 (13.0, 46.5) min, and the maximum time gap was 135 min. The 20 enrolled inpatients aged (45±12) years, including 15 males (75.0%). There was no significant difference in the blood concentration of tacrolimus collected at 7∶30 on the second (7.87±2.21)ng/ml and third days (7.84±2.33)ng/ml after admission of the enrolled inpatients (P=0.917), and the blood tacrolimus concentration rhythm was stable in the trial. The plasma concentration of C10.5-C14.5 was linearly related to the time, with R2 [M (Q1, Q3)] 0.88 (0.85, 0.92) and all P<0.05. The metabolic rate of tacrolimus during C10.5-C14.5=0.984+0.090×basic concentration of tacrolimus (ng/ml)-0.036×body mass index+0.489×CYP3A5 genotype-0.007×hemolobin(g/L)-0.035×alanine aminotransferase (U/L)+0.143×total cholesterol (mmol/L)+0.027×total bilirubin (µmol/L), with R2=0.85. Conclusion: This study propose a correction model for tacrolimus (non-sustained-release dosage form) trough concentration around C12, which is helpful for clinicians to easily and accurately assess renal transplant recipients' tacrolimus exposure.

[Functional characteristics of YAP-positive hepatocytes expression in an early stage of NASH with transcriptome sequence analysis].

Objective: To analyze and compare the differentially expressed genes (DEGs) of Yes-associated protein (YAP)-positive and negative hepatocytes and further understand the preliminary functional characteristics of YAP-positive hepatocytes in an early mouse model of nonalcoholic steatohepatitis (NASH) with transcriptome sequence (RNA-Seq). Methods: C57BL/6 mice were fed with methionine-choline deficiency (MCD) diet for 2 weeks to establish an early NASH model, and the control group was fed with normal diet. Liver tissue was stained with hematoxylin-eosin (HE) and Sirius red, and the pathological score was recorded. The expression of YAP and P-YAP were determined by immunohistochemistry (IHC) in liver tissues. Primary hepatocytes with viability greater than 90% were isolated and purified by collagenase perfusion combined with Percoll density gradient centrifugation. YAP-positive and negative hepatocytes were assessed by YAP antibody, flow cytometry and RNA-Seq analyses. Sequencing results were screened by GO, KEGG and interaction network analysis methods. RT-PCR was used to verify the expression levels of YAP and some DEGs in liver tissue model group. Two samples mean was compared by independent samples t-test. Results: Compared with the control group, the HE-stained liver tissue of MCD-induced mice at 2 weeks showed steatosis (pathological score 1.07±0.21), accompanied by lobular inflammation (pathological score 1.13±0.32) and ballooned hepatocyte (pathological score 0.80) ±0.20). Sirius red staining showed non-significant liver fibrosis (pathological score 0.40±0.40). IHC showed partial YAP-positive hepatocytes expression in an early stage of NASH. RNA-Seq analysis showed that clean reads of YAP-positive and negative hepatocytes were 49 310 604 and 5 4820 036, respectively. Compared with YAP-negative hepatocytes, YAP-positive hepatocytes had differential expression of 5 565 genes, including 1 662 up-regulated genes and 3 903 down-regulated genes. GO analysis of up-regulated genes showed that the metabolic processes related to mitochondrial functions, such as purine nucleoside triphosphate and nucleoside triphosphate were significantly enriched in biological processes (BP), while down-regulated gene analysis showed that olfactory-related receptor were significantly enriched in BP. KEGG analysis showed that DEGs were enriched in 292 pathways, and oxidative phosphorylation (OXPHOS) pathway was significantly enriched in signaling pathway. RT-PCR validated that inflammatory factors (interleukin-1ß, interleukin-6), YAP and its target genes (Cyr61, Ankrd1), and Cox5b and Sdhc genes were significantly up-regulated in the OXPHOS pathway, which was consistent with the sequencing results. In addition, eight key genes with interaction network analysis were predicted. Conclusion: Changes in hepatocyte metabolic levels may be associated with increased YAP activity in an early stage of NASH.

[A study on the identification of threshold for early warning on adverse weather events based on the association of apparent temperature and years of life lost].

Objective: To identify the threshold of a health warning system based on the association of apparent temperature and years of life lost (YLL). Methods: Daily mortality records and meteorological data were collected from 364 Chinese counties for 2006-2017. Distributed lag nonlinear model and multivariate Meta-analyses were applied to estimate the association between the apparent temperature and YLL rate. A regression tree model was employed to estimate the warning thresholds of the apparent temperature. Stratified analyses were further conducted by age and cause of death. Results: The daily YLL rate was 23.6/105. The mean daily apparent temperature was 15.7 ℃. U-shaped nonlinear associations were observed between apparent temperature and YLL rate. The actual temperature-caused YLL rate for the elderly was higher than the young population. The daily excess deaths rate increased with the higher effect levels. Conclusions: Regression tree model was employed to define the warning threshold for meteorological health risk. The present study provides theoretical support for the weather-related health warning system.

[Comparison of two epidemic patterns of COVID-19 and evaluation of prevention and control effectiveness: an analysis based on Guangzhou and Wenzhou].

Objective: To compare the epidemiological characteristics of COVID-19 in Guangzhou and Wenzhou, and evaluate the effectiveness of their prevention and control measures. Methods: Data of COVID-19 cases reported in Guangzhou and Wenzhou as of February 29, 2020 were collected. The incidence curves of COVID-19 in two cities were constructed. The real time reproduction number (R(t)) of COVID-19 in two cities was calculated respectively. Results: A total of 346 and 465 confirmed COVID-19 cases were analysed in Guangzhou and Wenzhou, respectively. In two cities, most cases were aged 30-59 years (Guangzhou: 54.9%; Wenzhou: 70.3%). The incidence curve peaked on 27 January, 2020 in Guangzhou and on 26 January, 2020 in Wenzhou, then began to decline in both cities. The peaks of imported COVID-19 cases from Hubei occurred earlier than the peak of COVID-19 incidences in two cities, and the peak of imported cases from Hubei occurred earlier in Wenzhou than in Guangzhou. In early epidemic phase, imported cases were predominant in both cities, then the number of local cases increased and gradually took the dominance in Wenzhou. In Guangzhou, the imported cases was still predominant. Despite the different epidemic pattern, the R(t) and the number of COVID-19 cases declined after strict prevention and control measures were taken in Guangzhou and in Wenzhou. Conclusion: The time and scale specific differences of imported COVID-19 resulted in different epidemic patterns in two cities, but the spread of the disease were effectively controlled after taking strict prevention and control measures.

[Risk assessment and early warning of imported COVID-19 in Guangdong province].

Objective: To assess the imported risk of COVID-19 in Guangdong province and its cities, and conduct early warning. Methods: Data of reported COVID-19 cases and Baidu Migration Index of 21 cities in Guangdong province and other provinces of China as of February 25, 2020 were collected. The imported risk index of each city in Guangdong province were calculated, and then correlation analysis was performed between reported cases and the imported risk index to identify lag time. Finally, we classified the early warming levels of epidemic by imported risk index. Results: A total of 1 347 confirmed cases were reported in Guangdong province, and 90.0% of the cases were clustered in the Pearl River Delta region. The average daily imported risk index of Guangdong was 44.03. Among the imported risk sources of each city, the highest risk of almost all cities came from Hubei province, except for Zhanjiang from Hainan province. In addition, the neighboring provinces of Guangdong province also had a greater impact. The correlation between the imported risk index with a lag of 4 days and the daily reported cases was the strongest (correlation coefficient: 0.73). The early warning base on cumulative 4-day risk of each city showed that Dongguan, Shenzhen, Zhongshan, Guangzhou, Foshan and Huizhou have high imported risks in the next 4 days, with imported risk indexes of 38.85, 21.59, 11.67, 11.25, 6.19 and 5.92, and the highest risk still comes from Hubei province. Conclusions: Cities with a large number of migrants in Guangdong province have a higher risk of import. Hubei province and neighboring provinces in Guangdong province are the main source of the imported risk. Each city must strengthen the health management of migrants in high-risk provinces and reduce the imported risk of Guangdong province.

[Risk assessment of exported risk of COVID-19 from Hubei Province].

Objective: To evaluate the exported risk of COVID-19 from Hubei Province and the imported risk in various provinces across China. Methods: Data of reported COVID-19 cases and Baidu Migration Indexin all provinces of the country as of February 14, 2020 were collected. The correlation analysis between cumulative number of reported cases and the migration index from Hubei was performed, and the imported risks from Hubei to different provinces across China were further evaluated. Results: A total of 49 970 confirmed cases were reported nationwide, of which 37 884 were in Hubei Province. The average daily migration index from Hubei to other provinces was 312.09, Wuhan and other cities in Hubei were 117.95 and 194.16, respectively. The cumulative COVID-19 cases of provinces was positively correlated with the migration index derived from Hubei Province, also in Wuhan and other cities in Hubei, with correlation coefficients of 0.84, 0.84, and 0.81. In linear model, population migration from Hubei Province, Wuhan and other cities in Hubei account for 71.2%, 70.1%, and 66.3% of the variation, respectively. The period of high exported risk from Hubei occurred before January 27, of which the risks before January 23 mainly came from Wuhan, and then mainly from other cities in Hubei. Hunan Province, Henan Province and Guangdong Province ranked the top three in terms of cumulative imported risk (the cumulative risk indices were 58.61, 54.75 and 49.62 respectively). Conclusion: The epidemic in each province was mainly caused by the importation of Hubei Province. Taking measures such as restricting the migration of population in Hubei Province and strengthening quarantine measures for immigrants from Hubei Province may greatly reduce the risk of continued spread of the epidemic.

[The definition of heat-wave based on mortality risk assessment in different regions of China].

Objective: To identify the definition of heat wave based on mortality risk assessment in different regions of China. Methods: Daily mortality (from China Information System for Disease Control and Prevention) and meteorological data (from National Meteorological Information Center in China) from 66 counties with a population of over 200 000 were collected from 2006-2011. With the consideration of climate type and administrative division, China was classified as seven regions. Firstly, distributed lag non-linear model (DLNM) was used to estimate community-specific effects of temperature on non-accidental mortality. Secondly, a multivariate meta-analysis was applied to pool the estimates of community-specific effects to explore the region-specific temperature threshold and the duration for definition of heat wave. Results: We defined regional heat wave of Northeast, North, Northwest, East, Central and Southwest China as being two or more consecutive days with daily mean temperature higher than or equal to the P(64), P(71), P(85), P(67), P(75) and P(77) of warm season (May to October) temperature, respectively, while the thresholds of temperature were 21.6, 23.7, 24.3, 25.7, 28.0 and 25.3 ℃. The heat wave in South China was defined as five or more consecutive days with daily mean temperature higher than or equal to the P(93) (30.4 ℃) of warm season (May to October) temperature. Conclusion: The region-specific definition of heat wave developed in our study may provide local government with the guidance of establishment and implementation of early heat-health response systems to address the negative health outcomes due to heat wave.

[Lagged effects of diurnal temperature range on mortality in 66 cities in China: a time-series study].

Objective: To estimate the effect of daily diurnal temperature range (DTR) on mortality in different areas in China. Methods: A time series study using the data collected from 66 areas in China was conducted, and Meta-analysis was used to analyze the estimates of associations between DTR and daily mortality. Modifying effects of extremely low and high DTR-mortality relationship by season and socioeconomic status (SES) were also evaluated respectively. Cumulative excess risk (CER) was used as an index to evaluate the effects. Results: The information about 1 260 913 registered deaths were collected between 1 January 2006 and 31 December 2011, we found the relationship between extreme DTR and mortality was non-linear in all regions and the exposure-response curve was J-shaped. In central and south areas of China, the result indicated the obvious acute effect of extremely high DTR, and the mortality effect in central area (CER=5.1%, 95% CI: 2.4%-7.9%) was significant higher than that in south area (CER=4.5%, 95% CI: 1.7%-7.3%). Regarding to the modification of seasons, the cumulative mortality effect of DTR in cold season (CER=5.8%, 95%CI: 2.5%-9.2%) was higher than that in hot season (CER=3.1%, 95%CI: 1.1%-5.1%). Generally, deaths among the elderly (≥75 years) were associated more strongly with extremely high DTR. Conclusions: The mortality effects of extremely DTR in different areas and seasons showed different characteristics, that in central area and in cold season it was significantly stronger. After modified by season and SES, DTRs were the greatest threat to vulnerable population, especially to the elderly (≥75 years). Therefore, more attention should be paid to vulnerable groups and protection measures should be taken according to the local and seasonal conditions.

[Commuting mode specific exposure to PM(2.5) in urban area of Guangzhou].

Objective: To find the differences in PM(2.5) exposure level in the context of four commuting modes (by walk, bicycle, bus and subway) in Guangzhou. Methods: The PM(2.5) exposure assessment was carried out from January to December 2015 in Guangzhou. PM(2.5) was measured by using SidePak individual dust meter (AM510, TSI Inc. USA) with time interval of 1 minute. Our measurement was taken on Monday, Wednesday, Friday and Sunday in the second week of each month and the samples were collected in the morning (07:00-09:00), afternoon (11:00-13:00) and evening (17:00-19:00). Results: A total of 284 air samples during walking, 281 air samples during bicycle riding, 278 air samples in bus, and 280 air samples in subway were collected. The median PM(2.5) concentrations exposed during walking, during bicycle riding, in bus and in subway were 38.4, 38.6, 23.3 and 24.1 µg/m(3), respectively, which were positive correlated with exposure concentration in fixed surveillance sites. The exposure level was lowest in summer, and highest in winter. The median of one-way exposure level to PM(2.5) from high to low were as follows: 21.0 µg for bicycle riding, 20.1 µg for walking, 5.1 µg for taking bus and 2.6 µg for taking subway. The season and time specific one-way exposure levels to PM(2.5) of four commuting modes were consistent. Conclusions: The exposure level to PM(2.5) was obviously higher during walking and bicycle riding than that in bus and subway. The exposure level to PM(2.5) during walking was higher than that during bicycle riding, in bus and in subway.

[Effectiveness of health education about heat wave hazard prevention in the elderly: a mixed effect model analysis].

Objective: To evaluate the effectiveness of health education about prevention of heat wave hazard in the elderly. Methods: A non-randomized controlled trial was conducted during the summer of 2015 among a sample of residents aged ≥60 years in Panyu district, Guangzhou. Eight intervention measures for heat wave hazard prevention were taken in intervention group for 3 months (from August to October) and in control group no intervention measures were taken. The comparison of intervention effects was conducted between the intervention group and control group with mixed effect model after the collection of related information with same questionnaire. Results: After adjusting of family per capita income, family air-condition availability, alcohol use, disease history and time, the average score of risk awareness in the intervention group increased by 1.62, while it was 0.51 in the control group, the difference was significant (t=2.76, P=0.006). A significant effect was observed in the intervention group on the reduction of hospitalizations due to chronic diseases. The hospitalization rate due to chronic diseases in resent 3 months in the intervention group decreased from 32.39% (46/142) before intervention to 28.87% (41/142) after intervention; while in the control group, it increased from 26.28% (41/156) before intervention to 36.53% (57/156) after intervention. There was no significant difference between the two groups in awareness of knowledge on heat wave hazard prevention and the score of adaptation to heat wave. Conclusion: Health education programs could improve the risk awareness on heat waves, and reduce the hospitalizations due to chronic diseases in the elderly.

[Studies on the growth characteristics of Spirulina cells under batch and continuous conditions].

The research paper focused mainly on the growth characteristics of Spirulina cells under batch and continuous conditions. It was found that the Richards and Exponential decay model could well express the relationships between the concentration of the cell, carbon in the medium and culture time respectively. The maximum cell growth rate and the cell yield coefficient against carbon under batch culture was 0.371 g/d/L and 3.439 g/gC respectively. The concentration of cell and that of carbon in the reactor increased and decreased with the dilution rate respectively under the continuous culture condition. The maximum cell growth rate and the optimum dilution rate was 0.362 g/d/L and 0.45/d respectively. The cell yield against carbon under continuous culture condition was 2.050 g/gC. The cell growth rate of batch culture was larger than that of continuous one when the cell concentration was relatively low, but it will be smaller than that of continuous culture when the cell concentration became high.

Many transcribed regions of the Onchocerca volvulus genome contain the spliced leader sequence of Caenorhabditis elegans.

Genomic DNAs of the related parasitic nematodes Onchocerca volvulus and Dirofilariae immitis, and a cDNA library of O. volvulus, were examined for the presence of the 22-nucleotide spliced leader (SL) found at the 5' ends of 10 to 15% of the mRNAs in the free-living nematode Caenorhabditis elegans. As in C. elegans, genes for the SL RNA are linked to the repetitive 5S rRNA genes of O. volvulus and D. immitis, but unlike C. elegans, they are in the same orientation as the 5S rRNA genes within the repeat unit. In O. volvulus the SL sequence is also encoded at more than 30 additional genomic locations and occurs at interior sites within many transcripts. Sequence determinations of four different cDNAs of O. volvulus, each containing an internal copy of the SL within a conserved 25mer, and one corresponding genomic DNA clone indicate that this sequence is not trans spliced onto these RNAs, but is encoded within the genes. The RNAs of two of these cDNAs appear to be developmentally regulated, since they occur in adult O. volvulus but were not detected in the infective L3 stage larvae. In contrast, actin mRNAs are present at all developmental stages, and at least one actin mRNA species contains a trans-spliced 5' SL. The internal locations of the SL in various transcripts and its perfect sequence conservation among parasitic and free-living nematodes argues that it serves specific, and perhaps multiple, functions for these organisms.

Construction of Onchocerca volvulus cDNA libraries and partial characterization of the cDNA for a major antigen.

Adult Onchocerca volvulus were isolated from nodules removed from onchocerciasis patients at four locations--two in the West African Sudan-savanna region (near Bamako, Mali, and Touboro, Cameroon), one in a West African forest region (Kumba, Cameroon) and one near Guatemala City, Guatemala. Four different cDNA expression libraries were constructed in bacteriophage lambda gt11 using poly(A)+ RNA from the adult female worms. Individual cDNA clones of single copy genes were used to compare the genomes of parasites from the different locales and to show that the haploid genome of O. volvulus is 1.5 x 10(8) base pairs. About 1 in 700 recombinant clones in each of the four amplified cDNA libraries produces a fusion protein recognized by pooled human anti-O. volvulus antisera. Partial sequence determination of a 2.0 kb cDNA clone for an O. volvulus protein that induces an immunodominant response in rabbits revealed that this antigen has sequence similarities with Caenorhabditis elegans myosin and with schistosome paramyosin (which confers partial protection against schistosome infection). The four cDNA libraries have been deposited with American Type Culture Collection (ATCC), 12301 Parklawn Drive, Rockville, MD 20852, U.S.A., for general distribution under ATCC Number 37509.