DNA methylation clocks for estimating biological age in Chinese cohorts.

Epigenetic clocks are accurate predictors of human chronological age based on the analysis of DNA methylation at specific CpG sites. However, available DNA methylation (DNAm) age predictors are based on datasets with limited ethnic representation. Moreover, a systematic comparison between DNAm data and other omics datasets has not yet been performed. To address these knowledge gaps, we generated and analyzed DNA methylation datasets from two independent Chinese cohorts, revealing age-related DNAm changes. Additionally, a DNA methylation (DNAm) aging clock (iCAS-DNAmAge) and a group of DNAm-based multi-modal clocks for Chinese individuals were developed, with most of them demonstrating strong predictive capabilities for chronological age. The clocks were further employed to predict factors influencing aging rates. The DNAm aging clock, derived from multi-modal aging features (compositeAge-DNAmAge), exhibited a close association with multi-omics changes, lifestyles, and disease status, underscoring its robust potential for precise biological age assessment. Our findings offer novel insights into the regulatory mechanism of age-related DNAm changes and extend the application of the DNAm clock for measuring biological age and aging pace, providing basis for evaluating aging intervention strategies.

MicroRNA-377-3p exacerbates chronic obstructive pulmonary disease through suppressing ZFP36L1 expression and inducing lung fibroblast senescence.

Chronic obstructive pulmonary disease (COPD) is a leading aging related cause of global mortality. Small airway narrowing is recognized as an early and significant factor for COPD development. Senescent fibroblasts were observed to accumulate in lung of COPD patients and promote COPD progression through aberrant extracellular matrix (ECM) deposition and senescence-associated secretory phenotype (SASP). On the basis of our previous study, we further investigated the the causes for the increased levels of miR-377-3p in the blood of COPD patients, as well as its regulatory function in the pathological progression of COPD. We found that the majority of up-regulated miR-377-3p was localized in lung fibroblasts. Inhibition of miR-377-3p improved chronic smoking-induced COPD in mice. Mechanistically, miR-377-3p promoted senescence of lung fibroblasts, while knockdown of miR-377-3p attenuated bleomycin-induced senescence in lung fibroblasts. We also identified ZFP36L1 as a direct target for miR-377-3p that likely mediated its pro senescence activity in lung fibroblasts. Our data reveal that miR-377-3p is crucial for COPD pathogenesis, and may serve as a potential target for COPD therapy.

TAF1B depletion leads to apoptotic cell death by inducing nucleolar stress and activating p53-miR-101 circuit in hepatocellular carcinoma.

Background: TAF1B (TATA Box Binding Protein (TBP)-Associated Factor) is an RNA polymerase regulating rDNA activity, stress response, and cell cycle. However, the function of TAF1B in the progression of hepatocellular carcinoma (HCC) is unknown. Objective: In this study, we intended to characterize the crucial role and molecular mechanisms of TAF1B in modulating nucleolar stress in HCC. Methods: We analyzed the differential expression and prognostic value of TAF1B in hepatocellular carcinoma based on The Cancer Genome Atlas (TCGA) database, tumor and paraneoplastic tissue samples from clinical hepatocellular carcinoma patients, and typical hepatocellular carcinoma. We detected cell proliferation and apoptosis by lentiviral knockdown of TAF1B expression levels in HepG2 and SMMC-7721 cells using clone formation, apoptosis, and Western blotting (WB) detection of apoptosis marker proteins. Simultaneously, we investigated the influence of TAF1B knockdown on the function of the pre-initiation complex (PIC) by WB, and co-immunoprecipitation (Co-IP) and chromatin immunoprecipitation (ChIP) assays verified the interaction between the complexes and the effect on rDNA activity. Immunofluorescence assays measured the expression of marker proteins of nucleolus stress, fluorescence in situ hybridization (FISH) assays checked the rDNA activity, and qRT-PCR assays tested the pre-rRNA levels. Regarding molecular mechanisms, we investigated the role of p53 and miR-101 in modulating nucleolar stress and apoptosis. Finally, the impact of TAF1B knockdown on tumor growth, apoptosis, and p53 expression was observed in xenograft tumors. Result: We identified that TAF1B was highly expressed in hepatocellular carcinoma and associated with poor prognosis in HCC patients. TAF1B depletion modulated nucleolar stress and apoptosis in hepatocellular carcinoma cells through positive and negative feedback from p53-miR-101. RNA polymerase I transcription repression triggered post-transcriptional activation of miR-101 in a p53-dependent manner. In turn, miR-101 negatively feeds back through direct inhibition of the p53-mediated PARP pathway. Conclusion: These findings broaden our comprehension of the function of TAF1B-mediated nucleolar stress in hepatocellular carcinoma and may offer new biomarkers for exploring prospective therapeutic targets in HCC.

Efficacy of Qizi Yusi Pill on Pregnancy Outcomes in Women of Advanced Reproductive Age: A Multicenter, Randomized, Double-Blind, Placebo-Controlled Trial.

OBJECTIVE: To evalvate efficacy of Qizi Yusi Pills (QYP), a Chinese medicine compound preparation, on in vitro fertilization-embryo transfer (IVF-ET) in women of advanced reproductive age. METHODS: This multicenter, randomized, double-blind, placebo-controlled trial was conducted from June 2018 to October 2019. A total of 124 patients were randomly allocated to either the QYP group or the placebo group using a stratified block randomization design, with 62 patients in each group. All patients completed controlled ovarian stimulation using a standard gonadotropin-releasing hormone agonist (GnRH-a) long protocol. As the QYP group, QYP was administered while the control group received placebo. QYP and placebo were administered for a total of 24 to 30 days from the day of GnRH-a pituitary downregulation to transvaginal oocyte retrieval. Both medications were taken orally at doses of 10 g three times each day. The primary outcome was cumulative pregnancy rate, and the secondary outcomes were periodic medication, follicular status, serum hormone and endometrial receptivity. Follow-up continued until 4 weeks after delivery. Maternal and neonatal complications, such as gestational diabetes, were also observed. RESULTS: Overall, 119 patients completed the study, 60 in the QYP group and 59 in the placebo group. Per protocol (PP) analysis revealed that 6-month cumulative pregnancy rate in the QYP group was significantly higher than that in the placebo group [43.33% (26/60) vs. 25.42% (15/59), P=0.040). Additionally, more oocytes were retrieved from the QYP group than those from the placebo group (8.95 ± 3.12 vs. 7.85 ± 1.91, P=0.022). Moreover, the endometrial thickness of HCG day in the QYP group was significantly higher than that in the placebo group (11.78 ± 2.27 mm vs. 10.68 ± 2.07 mm, P=0.012). Maternal and neonatal complications between the two groups were not significantly different (P>0.05). Intention-to-treat analysis was in line with PP results. CONCLUSIONS: QYP can enhance ovarian reserve capacity and ovarian response, and possibly promote endometrial receptivity. QYP effectively improves cumulative pregnancy rates in older patients (⩾35 years) undergoing IVF-ET. (Registration No. ChiCTR1800014427).

Is Human Chorionic Gonadotropin Trigger Beneficial for Natural Cycle Frozen-Thawed Embryo Transfer?

Objective: The aim of this study is to investigate, in ovulatory patients, whether there is a difference in reproductive outcomes following frozen-thawed embryo transfer (FET) in natural cycles (NC) compared to modified natural cycles (mNC). Methods: This retrospective cohort study, performed at the public tertiary fertility clinic, involved all infertile patients undergoing endometrial preparation prior to FET in NC and mNC from January, 2017 to November, 2020. One thousand hundred and sixty-two patients were divided into two groups: mNC group (n = 248) had FET in a NC after ovulation triggering with human chorionic gonadotropin (hCG); NC group (n = 914) had FET in a NC after spontaneous ovulation were observed. The primary outcome was live birth rate. All pregnancy outcomes were analyzed by propensity score matching (PSM) and multivariable logistic regression analyses. Results: The NC group showed a higher live birth rate [344/914 (37.6%) vs. 68/248 (27.4%), P = 0.003; 87/240 (36.3%) vs. 66/240 (27.5%), P = 0.040] than the mNC group before and after PSM analysis. Multivariable analysis also showed mNC to be associated with a decreased likelihood of live birth compared with NC [odds ratio (OR) 95% confidence interval (CI) 0.71 (0.51-0.98), P = 0.039]. Conclusion: For women with regular menstrual cycles, NC-FET may have a higher chance of live birth than that in the mNC-FET cycles. As a consequence, it's critical to avoid hCG triggering as much as possible when FETs utilize a natural cycle strategy for endometrial preparation. Nevertheless, further more well-designed randomized clinical trials are still needed to determine this finding.

Immediate versus delayed frozen embryo transfer in women following a failed IVF-ET attempt: a multicenter randomized controlled trial.

BACKGROUND: The optimal time at which to perform a frozen-thawed embryo transfer (FET) following a failed in-vitro fertilization-embryo transfer (IVF-ET) attempt remains elusive to most reproductive experts. Physicians often delay the introduction of FET due to concerns related to potential residual effects of ovarian hyperstimulation which may interfere with the regular menstrual cycle. Moreover, given that most of the published studies on the topic are retrospective and have inconsistent findings, it is crucial to develop evidence-based randomized control guides for clinical practice. Therefore, this well-designed randomized controlled trial (RCT) was conducted to determine whether it is necessary to delay FET for at least one menstrual cycle after the failure of fresh embryo transfer. METHODS: Infertile women eligible for IVF-ET were invited to participate in this multicenter, randomized, non-inferiority, parallel-group, unblinded, controlled trial at the academic fertility centers of four public hospitals in Chinese Mainland. Infertile women scheduled to receive their first FET cycle after a failed IVF-ET attempt were randomly assigned to either (a) the immediate FET group in which FET was performed in the first menstrual cycle following the failed IVF-ET cycle (n = 366) or (b) the delayed FET group in which FET was performed in the second or subsequent menstrual cycle following the failed IVF-ET cycle (n = 366). All FET cycles were performed during hormone replacement cycles for endometrial preparation. The primary outcome was the ongoing pregnancy, defined as a detectable fetal heart beat beyond twelve weeks of gestation. Secondary outcomes were other pregnancy-related outcomes, maternal and neonatal complications. Analysis was performed by both intention-to-treat and per-protocol principles. RESULTS: A total of 646 FETs were completed. The frequency of moderate to severe depression and high stress level prior to FET in delayed FET group were significantly higher than that in immediate FET group (10.6% vs 6.1%, p = 0.039; 30.3% vs 22.4%, p = 0.022, respectively). Immediate FET resulted in a higher frequency of clinical pregnancy than did delayed FET (41.7% vs 34.1%), for a relative risk (RR) of 1.23 (95% confidence interval [CI], 1.00-1.50; p = 0.045). Women who underwent immediate FET also had a lower frequency of biochemical pregnancy loss (11.7% vs. 30.6%), with a RR of 0.28 (95% CI 0.23-0.63, p < 0.001), and a higher frequency of embryo implantation (25.2% vs. 20.2%), with a RR of 1.25 (95% CI 1.01-1.53; p = 0.038). Although the ongoing pregnancy and live birth rates did not differ significantly between the immediate FET and delayed FET groups (37.1% vs 30.3%, RR 1.22, 95% CI 0.99-1.52, p = 0.067; 36.5% vs 30.0%, RR 1.22, 95% CI 0.98-1.52, p = 0.079, respectively), a multivariate logistic regression analysis adjusted for potential confounders such as depression and stress levels revealed that the immediate FET group had a significantly higher ongoing pregnancy and live birth rates than the delayed FET group (odds ratio 0.68, 95% CI 0.47-0.99, p = 0.041; odds ratio 0.67, 95% CI 0.46-0.96, p = 0.031). The risks of maternal and neonatal complications were comparable between the two groups. CONCLUSIONS: In women with a previous failed IVF-ET attempt, immediate FET resulted in higher ongoing pregnancy and live birth rates than delayed FET. These findings warrant caution in the indiscriminate application of a delayed FET strategy when apparent risk of high stress level is perceived. TRIAL REGISTRATION: ChiCTR2000033313 .

Unusual matrine-adenine hybrids isolated from Sophora davidii and their inhibitory effects on human cytomegalovirus.

A phytochemical investigation on the flowers of Sophora davidii resulted in the isolation of three unusual matrine-adenine hybrids, sophovicines A-C, together with biogenetically related analogue sophocarpine. Their structures and absolute configurations were determined by NMR analysis, single-crystal X-ray diffraction, and electronic circular dichroism (ECD) data. Since sophovicines represent the first example of matrine-adenine hybrids, a putative biosynthetic pathway toward sophovicines A-C was proposed. In addition, computational molecular modeling suggested that compounds sophovicines B and C may have potent activities against human cytomegalovirus (HCMV). So, the inhibit effects of isolates on HCMV were evaluated. The results show that sophovicines B and C can inhibit HCMV replication effectively with IC50 values of 7.12 and 7.32 µM, respectively.

Comparison of Stimulated Cycles with Low Dose r-FSH versus Hormone Replacement Cycles for Endometrial Preparation Prior to Frozen-Thawed Embryo Transfer in Young Women with Polycystic Ovarian Syndrome: A Single-Center Retrospective Cohort Study from China.

OBJECTIVE: The principal purpose of this study was to compare reproductive outcomes for stimulated cycles (STC) and hormone replacement cycles (HRC) for endometrial preparation before frozen-thawed embryo transfer (FET) in young women with polycystic ovary syndrome (PCOS). METHODS: We conducted a retrospective study of 1434 FET cycles from January, 2017 to March, 2020 in our reproductive center, in which stimulated and hormone replacement cycles were used for endometrial preparation. Pregnancy outcomes of couples undergoing routine STC-FET or HRC-FET were analyzed by propensity score matching (PSM) and multivariable logistic regression analyses. RESULTS: Data on 1234 HRC protocols (86% of the total) and 200 STC protocols (14%) were collected. After PSM, 199 patients were included in both groups, respectively. There was no significant difference in positive pregnancy rate (52.7% vs 54.8%, p=0.763), clinical pregnancy rate (51.8% vs 52.8%, p=0.841), live birth rate (45.2% vs 43.7%, p=0.762), pregnancy loss rate (9.7% vs 16.2%, p=0.164) and ectopic pregnancy rate (1.5% vs 0.5%, p=0.615) between STC and HRC protocols. Subsequent multivariate logistic regression analysis also yielded similar results. CONCLUSION: STC for endometrial preparation had similar pregnancy outcomes compared with HRC protocols. Evidence is available which shows that for young women with PCOS in preparation for FET, HRC could be a reasonable choice for patients who are unwilling to accept injections. However, STC may reduce unnecessary anxiety and operational costs and offer more flexibility for patients. Eventually, we must embrace the concepts of individualization, securitization, and optimization in the clinic.

The Role of Traditional Chinese Formula Ding-Kun Pill (DKP) in Expected Poor Ovarian Response Women (POSEIDON Group 4) Undergoing In Vitro Fertilization-Embryo Transfer: A Multicenter, Randomized, Double-Blind, Placebo-Controlled Trial.

Objective: The primary objective of the study was to assess traditional Chinese formula DKP supplementation in terms of efficacy and safety on reproductive outcomes of expected poor ovarian responder (POR, POSEIDON Group 4) undergoing in vitro fertilization-embryo transfer (IVF-ET). Design Setting and Participants: Women eligible for IVF-ET were invited to participate in this randomized, double-blind, placebo-controlled, superiority trial at academic fertility centers of ten public hospitals in Chinese Mainland. A total of 462 patients (35-44 years) equally divided between DKP and placebo groups with antral follicle count (AFC) <5 or anti-müllerian hormone (AMH) <1.2 ng/ml were randomized. Interventions: All participants were given DKP or 7 g placebo twice daily on the previous menstrual cycle day 5 until oocyte retrieval, which took approximately 5 to 6 weeks. Main Outcome Measure: The primary outcome was the ongoing pregnancy defined as more than 20 gestational weeks of an intrauterine living fetus confirmed by pelvic ultrasonography. Results: Demographic characteristics were equally distributed between the study populations. Intention-to-treat (ITT) analysis revealed that ongoing pregnancy rate (OPR) was not significantly different between DKP and placebo groups [26.4% (61/231) versus 24.2% (56/231); relative risk (RR) 1.09, 95% confidence interval (CI) 0.80 to 1.49, P = 0.593]. No significant differences between groups were observed for the secondary outcomes. The additional per protocol (PP) analysis was in line with ITT results: OPR in DKP group was 27.2% (61/224) versus 24.1% (55/228) in placebo group [RR 1.13, 95%CI (0.82 to 1.55), P = 0.449]. After subgroup analysis the findings concluded that POR population of 35-37 years had a significantly higher OPR after 5-6 weeks of oral DKP (41.8%, 33/79) versus placebo (25.4%, 18/71) [RR 1.65, 95% CI (1.02 to 2.65), P = 0.034, P for interaction = 0.028]. Conclusion: This well-designed randomized controlled trial (RCT) offers new high-quality evidence to supplement existing retrospective literature concerning DKP performance in expected PORs. DKP could be recommended as a safe and natural remedy for expected PORs (aged 35-37 years) who fulfill the POSEIDON group 4 criteria. However, additional interventional clinical studies are undoubtedly required to be conducted in the future to validate this hypothesis. Clinical Trial Registration: www.chictr.org.cn, identifier ChiCTR1900026614.

[Vehicle Air Pollutant Emission Inventory and Characterization in Henan Province from 2016 to 2019].

Based on the collected urban motor vehicle activity ownership and traffic flow of highways, combined with the mileage and source profiles of VOCs, using the emission factor method, we established high-resolution emission inventories from 2016 to 2019 for urban and 2016-based highway motor vehicles, respectively, in Henan Province, China. The results showed that gasoline vehicles, particularly minibuses and ordinary motorcycles, were the main contributors of CO, VOCs, and NH3, whereas heavy-duty and light-duty diesel trucks emitted SO2, NOx, and PM. Vehicles with China 1, China 3, and China 4 emission standards contributed significantly to pollutant emissions in the fleet. The temporal variation in traffic flow was consistent with the changes in freight and passenger traffic, with higher coefficients of variation for highways from August to October and the lowest in November. The weekly and daily changes in urban trunk roads showed distinct weekend effects and clear double-peak features, respectively. High-value emission areas were concentrated in urban centers with dense transport networks and high traffic volumes and on roads radiating outward from urban areas. The Lianhuo Expressway and the Beijing-Hong Kong-Macau Expressway were high-emission roads. Light-duty gasoline vehicles made the largest contribution to the ozone formation potential (OFP) of VOCs from motor vehicles. Five species, such as ethylene and propylene, contributed significantly to VOC emissions and OFP. The average annual growth rate of vehicle ownership from 2016 to 2019 was 5.7%. Compared with 2016, VOC emissions increased by 2.8% in 2019, whereas emissions of other pollutants showed decreasing trends of different degrees, with decreases of 76.3%, 51.7%, 50.3%, 43.1%, 16.7%, and 5.9% for SO2, PM2.5, PM10, NH3, CO, and NOx, respectively. The emission reduction percentage of each pollutant in 2019 under the control policies relative to the baseline scenario ranged from 15.6% to 82.4%.

[Differential Responses of Rhizospheric nirK- and nirS-type Denitrifier Communities to Different Phosphorus Levels in Paddy Soil].

Phosphorus is an essential life element, which can affect the activities and functions of denitrifiers. Both nirK and nirS genes can code nitrite reductase; however, it remains unclear whether nirK- and nirS-containing denitrifers respond differentially to changes in the availability of phosphorus in paddy soil. In this study, P-deficient paddy soil was used to grow rice plants. Three phosphorus levels established by applying P fertilizer at a rate of 0 mg·kg-1 (CK), 15 mg·kg-1 (P1), and 30 mg·kg-1(P2), respectively. The abundance and community structure of nirK- and nirS- containing denitrifers were determined using quantitative PCR and high-throughput sequencing techniques. Results indicated that nirK- and nirS-containing communities responded differentially to changes in the P levels. The nirS-containing communities are more sensitive to the changes in P in both rhizosphere and bulk soil samples. In addition, the abundance of nirS genes was 2-3 times higher in the P2 treatment than in the CK treatment. Furthermore, the nirS community structure is also clearly differed from the CK treatment. However, P addition only induced partial modification of the community structure and abundance of nirK-containing denitrifiers. Moreover, compared to the bulk soil with each phosphorus level, the nirS community structure in the rhizosphere soil changed significantly; however, only the P2 treatment induced significant increases in the abundance of the nirS gene. In contrast, no significant differences in the abundance and composition of nirK-containing denitrifers were detected between rhizosphere and bulk soils under different phosphorus levels. Collectively, application of phosphate fertilizer in P-deficient paddy soil could significantly increase the abundance of nirK- and nirS-containing denitrifiers, changing their community structures, with nirS-type showing a greater sensitivity than nirK-type denitrifiers. In comparison, the denitrifying communities in the rhizosphere were more sensitive to variable P levels than that in the bulk soil. Compared to bulk soils, rice growth shifted the community structure of nirS- and nirK-containing denitrifiers in rhizosphere soils at each level of P, but failed to induce significant changes in their abundance (except for P2) that could cause a significant increase in nirS abundance. These results could provide a theoretical basis for exploring the effects of fertilization on soil denitrification.

Gene expression profiling of brain metastatic cell from triple negative breast cancer: Understanding the molecular events.

Brain metastatic triple negative breast cancer (BM-TNBC) is afflicted with unfavorable prognosis. However, the molecular events underlying BM-TNBC remain largely unknown. In the present study, we conducted gene expression microarray analysis using the triple negative breast cancer cell line MDA-MB-231 and its brain metastatic derivative (MDA-MB-231Brm). Results of microarray analysis showed that a total of 4296 genes were differentially expressed, of which 2433 genes were up-regulated and 1863 genes were down-regulated. Gene Ontology (GO), KEGG pathway and protein-protein interaction (PPI) analyses indicated differentially expressed genes functionally categorized as genes of signal transduction, multicellular organismal development, ion transport, nervous system development, plasma membrane, extracellular region, calcium ion binding, GTP binding neuroactive ligand-receptor interaction. The validity of the microarray results was verified by quantitative real-time PCR analysis of twelve representative genes. The present findings revealed molecular basis and events associated with brain metastasis in TNBC, which will potentially contribute to the understanding of underlying mechanism and develop therapeutic targets.

Recombinant adenovirus containing hyper-interleukin-6 and hepatocyte growth factor ameliorates acute-on-chronic liver failure in rats.

AIM: To investigate the protective efficacy of recombinant adenovirus containing hyper-interleukin-6 (Hyper-IL-6, HIL-6) and hepatocyte growth factor (HGF) (Ad-HGF-HIL-6) compared to that of recombinant adenovirus containing either HIL-6 or HGF (Ad-HIL-6 or Ad-HGF) in rats with acute-on-chronic liver failure (ACLF). METHODS: The recombinant adenoviruses containing HIL-6 and/or HGF were constructed. We established an ACLF model, and rats were randomly assigned to control, model, Ad-GFP, Ad-HIL-6, Ad-HGF or Ad-HGF-HIL-6 group. We collected serum and liver tissue samples to test pathological changes, biochemical indexes and molecular biological indexes. RESULTS: Attenuated alanine aminotransferase, prothrombin time, high-mobility group box 1 (HMGB1), endotoxin, tumour necrosis factor (TNF)-α and interferon-γ were observed in the Ad-HGF-, Ad-HIL-6- and Ad-HGF-HIL-6-treated rats with ACLF. Likewise, reduced hepatic damage and apoptotic activity, as well as reduced HMGB1 and Bax proteins, but raised expression of Ki67 and Bcl-2 proteins and Bcl-2/Bax ratio were also observed in the Ad-HGF-, Ad-HIL-6- and Ad-HGF-HIL-6-treated rats with ACLF. More significant changes were observed in the Ad-HGF-HIL-6 treatment group without obvious side effects. Furthermore, caspase-3 at the protein level decreased in the Ad-HIL-6 and Ad-HGF-HIL-6 treatment groups, more predominantly in the latter group. CONCLUSION: This study identifies that the protective efficacy of Ad-HGF-HIL-6 is more potent than that of Ad-HGF or Ad-HIL-6 in ACLF rats, with no significant side effects.

Treatment of recurrent epileptic seizures in patients with neurological disorders.

This study aimed to investigate the clinical characteristics and the treatment principles and methods of recurrent epileptic seizures in patients with neurological disorders. A retrospective analysis was performed of the clinical data, treatment methods and results in 13 patients with recurrent epileptic seizures attending the neurosurgery department. Of the 13 patients, 10 had a history of epilepsy, 9 had organic frontal lobe brain lesions and 11 exhibited frontal lobe epilepsy. The causes of the epileptic seizure aggravation included drug withdrawal, dose reduction and dressing change (5 cases). The epileptic seizure types included partial and secondary full seizures and the seizure frequency ranged from 1 seizure/3 min to 1 seizure/several h. Following combined therapy with multiple anti-epileptic drugs (AEDs), including oral administration and injection, the epilepsy was controlled. The addition of orally administered levetiracetam improved the treatment efficiency. In cases of recurrent epileptic seizures in patients with neurological disorders, the combined administration of AEDs should be conducted with doses higher than the conventional initial dose to control the epileptic seizures as rapidly as possible.

[Expression of helicobacter pylori alpA gene in lactococcus lactis and its immunogenicity analysis].

AIM: To express Helicobacter pylori (Hp) alpA gene in a live delivery vehicle of lactococcus lactis (L. lactis) and assay the efficacy of the L. lactis-alpA oral vaccine in recipient mice. METHODS: The alpA gene of Hp was amplified by PCR and cloned into the prokaryotic expression vector pNICE: sec. The recombinant vector pNICE: sec-alpA was transformed into Lactococcus lactis strain NZ9000. Then the engineered strain was induced to express recombinant alpA as shown by SDS-PAGE and Western blot. Female ICR mice (CV Grade) were randomly divided into 4 groups and administrated orally with PBS, L. lactis pNICE: sec, L. lactis pNICE: sec-alpA, and the inactivated Hp, respectively. After immunized seven times, the mice were detected for their alpA-specific IgG and IgA. RESULTS: The alpA gene was obtained and successfully cloned into the vector pNICE: sec. The recombinant alpA protein (56,000) was accumulated in L. lactis after the induction of the nisin, accounting for 9.6% of the total bacterial protein. Western bolt confirmed that the alpA protein could be recognized specifically by the anti-Hp serum. The titer of anti-alpA IgG in the pNICE: sec-alpA group, comparable to that in the inactivated Hp group, was higher than that in the pNICE: sec group. The titer of anti-alpA IgA in the pNICE: sec-alpA group was higher than all other groups (P<0.05). CONCLUSION: The oral administration of the engineered alpA-expressing L. lactis induced protective immunity against Hp. Our study provides a certain experimental basis for the use of L. lactis as an antigen-delivering vehicle for the development of Hp oral vaccines.

[Vertical distribution characteristics and composition of saturated hydrocarbon in soil profiles of Beijing].

Soil samples were collected from ten soil profiles with different environmental conditions in Beijing for saturated hydrocarbons (SHs) analysis. The vertical distribution of the both concentration and composition of SHs in ten soil profiles were investigated. Concentration of SHs in different profiles is significantly different, ranged from 1.5 microg x g(-1) to 54.1 microg x g(-1). The higher concentrations are found in the samples from B7, B9 and B10. A series of SHs including n-alkanes, isoprenoid alkanes, terpenoids and steranes, alkyl hexamethylene were detected in all samples. The order from higher content to lower in most profiles (except B7) is: n-alkanes, isoprenoid alkanes, terpenoids and steranes, alkyl hexamethylene, and the relative content of n-alkanes is obviously dominant in the upper horizons. Concentrations of SHs and content of soil organic carbon in each profile show similar trend with depth, declined rapidly down to 30 cm and trend consistent in the deep part( > 40 cm). CPI1, CPI2, (C25 + C27 + C29 + C31) / Sigma alkanes, terpenoids and steranes, and biomarker parameters suggest that pollutants are mainly from fossil fuel in B7, high plant-derived n-alkanes dominated in other profiles and fossil fuel contamination to different extent. The sources of n-alkanes in deep part of soil profiles are different from those in topsoil samples, which are related to the soil itself, such as soil parent material and process of soil formation, but the sources of alkyl hexamethylene, terpenoids and steranes between topsoil and deep part are constant.