The effect of cold exposure on the levels of glucocorticoids, 11-hydroxysteroid dehydrogenase 2, and placental vascularization in a rat model.

OBJECTIVE: Cold exposure (CE) before birth is one of the initial stressors that may impact mammalian pregnancy, changing placental and fetal development and affecting the health of the offspring. While glucocorticoids (GCs) participate in the body's response to the stress of CE, the specific mechanisms of their action are unclear. This study aims to determine the effect of CE stress on the placenta and to test whether stress, caused by cold exposure in pregnancy impairs fetal development by changing placental angiogenesis via excessive GC expression. MATERIALS AND METHODS: CE rat model was created by exposing 30 SD rats to cold preconception, or during the first, second, and third weeks of pregnancy. Serum cortisol and soluble fms-like tyrosine kinase-1 (sFlt-1) expression levels, physiological index changes (food intake, body weight change and blood pressure), and pregnancy outcomes (fetal rat weight, number of live fetal rats, and placental weight) were collected at baseline and at different time points after the conception. Protein expression levels of 11 ß-hydroxysteroid dehydrogenase 2 (11ß-HSD2), glucocorticoid receptor, vascular endothelial growth factor A (VEGF-A), placental growth factor (PIGF), and sFlt-1 in placental tissues were measured by western blotting. Cytokeratin (CK) and laminin (LN) in trophoblasts, and α-actin in vascular smooth muscle of the spiral arteries of pregnant rats after the systemic cold treatment were assessed by immunofluorescence and visualized by fluorescent microscopy. To test the effect of 11ß-HSD2 levels on the placental recasting, human first-trimester extravillous trophoblast cells (HTR8/SVneo) underwent knockdown using specific 11ß-HSD2 siRNA constructs.  Expression levels of 11ß-HSD2 were analyzed by quantitative real-time PCR (qPCR) and into HTR8 cells, and the expression levels of the 11ß-HSD2 gene in each group were measured using qPCR. Cell migration and invasion was assessed by Transwell migration assay, and sFlt-1 levels in HTR8 cells were measured by ELISA. RESULTS: CE pre-conception led to consistently increasing serum corticosterone and sFlt-1 levels throughout pregnancy, and persistently increased diastolic blood pressure (DBP) in rat CE model compared to control animals. CE during the second week of gestation (Gp.3) was associated with significantly lower placental weight (p=0.0003). Cold exposure in the third week (Gp.4) was associated with significantly (p=0.001) lower fetal weight. CE pre-conception was associated with significantly decreased placental levels of 11ß-HSD2, glucocorticoid receptor, VEGF-A, PIGF, and sFlt-1 proteins and α-actin compared to the control group. Silencing 11ß-HSD2 by siRNA led to reduced cell migrations and invasion, and markedly increased expression levels of sFlt-1 in HTR8/SVneo cells (p<0.05). CONCLUSIONS: Pre-conception cold exposure and during early pregnancy leads to increased GCs levels and impaired placental 11ß-HSD2 activity. We suggest that the subsequent 11ß-HSD2-induced increase in the sFlt-1expression during early pregnancy may affect placental vascular remodeling and change placental morphological structure and function.

A model for pregnancy rates after IVF-ET in patients with infertility and endometriosis.

OBJECTIVE: The objective of the present study was to examine the clinical factors influencing the pregnancy rate of infertile patients with endometriosis and establish a predictive model. PATIENTS AND METHODS: This study included 158 patients (158 cycles) with infertility and endometriosis who underwent laparoscopic surgery, and in vitro fertilization and embryo transfer (IVF-ET) were evaluated retrospectively between January 2019 and December 2020. The clinical factors in the pregnant and non-pregnant group were analyzed by univariate analysis. Statistically significant variables were subsequently used for multivariate logistic regression to establish the prediction model. RESULTS: Multivariate logistic regression analyses showed that GnRH-a treatment after operation (OR, 6.562; 95% CI: 2.782-15.477; p<0.01), ASRM stage (OR, 0.218; 95% CI: 0.093-0.509; p<0.05), the number of high-quality transferred embryos (OR, 3.155; 95% CI: 1.647-6.047; p<0.05) were independently associated with successful pregnancy. The area under the curve (AUC) of the prediction model was 0.774 (95% CI: 0.700-0.847). According to Hosmer-Lemeshow, the model was well fitted (p>0.05). We applied the bootstrapping method to internal validation, and the result showed that the pregnancy rate predicted by the model and the real data were consistent. CONCLUSIONS: The models for predicting pregnancy rates after IVF-ET in infertility and endometriosis patients showed high accuracy. The effective methods to increase the number of high-quality embryos need further study.

Effect of induction of labor on maternal and perinatal outcomes in low-risk singleton pregnancies: a retrospective case-control study.

OBJECTIVE: The aim of the study was to investigate the effect of induction of labor on maternal and fetal outcomes. PATIENTS AND METHODS: This retrospective case-control study included 4386 pregnant women with low-risk singleton pregnancies who underwent regular prenatal examination and successful vaginal delivery at ≥41 weeks and 0 days of gestation in Fujian Maternal and Child Health Hospital between January 2014 and December 2018. Clinical data of women were retrospectively divided into the induction of labor group (2007 cases) and spontaneous onset of labor group (2361 cases) based on the mode of labor initiation. Two-sample independent t-test and χ2 tests were used to analyze the differences in clinical characteristics, such as maternal age and parity between the two groups. RESULTS: The induction of the labor group had a significantly longer total duration of labor (9.37±5.37 vs. 8.82±5.13 h; p<0.001), more postpartum blood loss (219.18±188.32 vs. 199.95±124.69 mL; p=0.01) and a significantly higher incidence of severe postpartum hemorrhage (PPH) comparing to the spontaneous onset of labor group [0.8% (16/2007) vs. 0.33% (8/2361); p=0.041]. However, no significant difference was found in the neonatal outcomes. After adjusting for age, induction of labor in nulliparous women was more likely to lead to PPH than the spontaneous onset of labor [2.74% (55/2007) vs. 1.65% (39/2361); odds ratio=1.557; 95% confidence interval: 1.039-2.332; p<0.05]. CONCLUSIONS: Induction of labor increases the duration of labor and postpartum blood loss, especially in primary parturient, leading to an increased risk of PPH. This increase may be related to the overall higher duration of labor, associated with IOL. Therefore, low-risk nulliparous women should try to avoid induction of labor without medical indications.

Efficacy of robot-assisted core decompression combined with human umbilical cord-derived mesenchymal stem cell transplantation for osteonecrosis of the femoral head.

OBJECTIVE: The aim of this paper was to evaluate the effects of robot-assisted core decompression combined with human umbilical cord-derived mesenchymal stem cell (hUC-MSC) transplantation for early-stage osteonecrosis of the femoral head (ONFH). PATIENTS AND METHODS: A retrospective analysis was performed on 18 patients with a total of 26 hips who were diagnosed with Association Research Circulation Osseous stage 2 avascular necrosis of the femoral head and who received core decompression combined with hUC-MSC transplantation. All surgeries were completed under robotic assistance. Preoperative and postoperative visual analogue scale (VAS) scores and the Harris Hip Score (HHS) were recorded to assess clinical function. A Magnetic Resonance Imaging (MRI) examination was performed at the last follow-up. RESULTS: The mean follow-up was 18.6 months (12-28 months), the VAS score (4.5±0.8 vs. 0.9±0.2, t=12.6, p≤0.001) and HHS (79.5±5.8 vs. 60.5±4.6, t=14.3, p≤0.001) were significantly improved at the last follow-up, compared with preoperative value. The MRI results showed that the necrotic volume of the femoral heads was significantly reduced. CONCLUSIONS: Robot-assisted core decompression combined with hUC-MSC transplantation is a feasible and relatively safe method for the treatment of femoral head necrosis.

Analysis and prediction of risk factors of ovarian hyperstimulation caused by Long-acting GnRH agonist protocol in follicular phase.

OBJECTIVE: The aim of the study was to explore the risk factors of ovarian hyperstimulation in patients undergoing long-acting gonadotropin-releasing hormone (GnRH) agonist protocol in follicular phase of ovulation induction therapy and to establish a predictive model. PATIENTS AND METHODS: A total of 1289 patients who received Long-acting GnRH agonist protocol in follicular phase for ovulation induction in the Fujian Provincial Maternity and Child Health Hospital from July 1, 2018, to July 31, 2019, were selected. Among them, 33 patients developed moderate/severe ovarian hyperstimulation syndrome. The relevant indicators of the two groups were followed up for comparison, and Lasso regression was used to screen independent risk factors and construct a nomogram prediction model.  A receiver operating characteristic (ROC) curve and calibration curve were used to evaluate the discrimination and calibration of the prediction model. RESULTS: Univariate analysis suggested that the woman's age, basal antral follicle number (AFC), total gonadotropin (Gn) dose, Gn starting dose, basal estradiol (E2) level, basal anti-Müllerian hormone (AMH) value, number of follicles obtained, Gn start day E2, the difference in follicle-stimulating hormone (FSH) value and Gn starting day were statistically significant. Significant indicators of univariate analysis and clinical significance were included in the Lasso regression model, and AFC, woman's age, polycystic ovary syndrome, Gn starting dose and number of follicles obtained were finally screened as final predictors. The ROC curve indicated that the area under the curve (AUC) was 0.812. CONCLUSIONS: Ovarian hyperstimulation caused by long-acting GnRH agonist protocol in follicular phase for ovulation stimulation has a certain predictability. Paying attention to the patient's age, AFC, Gn starting dose, number of follicles obtained, and whether PCOS is evident may lead to early detection of ovarian hyperstimulation syndrome, which has clinical guiding significance.

A new and improved acetabular cup digital templating method and its clinical application.

BACKGROUND AND OBJECTIVE: Preoperative acetabular cup templating has an important auxiliary effect on hip surgery. The traditional acetabular cup templating method requires the measuring person to have some experience in total hip replacement (THA) surgery since the measurement results vary from person to person with differences between different measuring persons. To obtain stable templating results, we designed a new acetabular cup templating method and tested the inter-person measuring differences and measurement accuracy of this method. Meanwhile, the clinical application of this method was preliminarily explored. MATERIALS AND METHODS: The pattern of this new method was manual labeling of imaging characteristic points and then programmed automatic measurements. The measurement process was performed entirely by orthopedic graduate students without any experience in hip replacement surgery. The inter-person measuring difference was evaluated by comparing the templating results of three measuring persons. The accuracy of the templating was evaluated by comparing the templating results with the actual size of the prosthesis in the surgery. The correlation between the position of the acetabular cup and the templating error was analyzed to explore the clinical significance of the templating results. This study was a retrospective study which included templating in a total of 406 cases for total hip replacement with cementless cup prosthesis. Digital measurements were performed using the Matlab software from MathWorks. The statistical comparison was performed using Kendall's W test. RESULTS: The results of the three measuring persons were completely identical in 61.8% (251/406) of cases, and the variation in 38.2% (155/406) of cases did not exceed one size of the acetabular cup. The Kendall's W coefficient was 0.977, and p < 0.01. The measurement accuracy is not as good as the traditional method in exactly accurate measurement and ±1 cup size, but it is similar to the traditional method in the ±2 cup sizes. The correlation between the templating error and the position evaluation of the implanted acetabular cups reveals: (1) larger the templating error, larger the proportion of the acetabular cups with poor position; (2) the proportion of acetabular cup with poor position slowly increased when the templating error was from 0 to 1 size, and the proportion rapidly increased when the templating error was from 1 to 2 size. CONCLUSION: All the patients with clear teardrop bottom and lateral superior edge of acetabulum were able to use our method to predict the size of the acetabular cup. The method has the following advantages: (1) it does not require the measuring person to have any previous experience of the THA surgery, which reduces the labor cost of the templating; (2) the differences between the measuring persons is small, the measurement result can be repeated; (3) it can predict the probability of acetabular cup with poor positioning according to the templating error, and thereby reminding the surgeon to recheck and correct the position of the acetabular cup in time during the surgery.

Scoring system to predict the success rate of external cephalic versions and determine the timing of the procedure.

OBJECTIVE: We aimed to examine the prenatal clinical characteristics of women with single pregnancies undergoing external cephalic version (ECV) without anesthesia, develop a novel scoring system for predicting the ECV success rate, and demonstrate that this scoring system can be used to individualize the timing of ECV attempts. PATIENTS AND METHODS: We enrolled 270 women who underwent ECV without anesthesia at 37-40 weeks of gestation in the Fujian Maternity and Child Health Hospital from 2016 to 2019 and divided them into two ECV outcome groups (success vs. failure). We identified five clinical features (the fetal buttocks' station, the sum of the fundal height and station, the fetal head location, and whether the fetal head or buttocks could be grasped) as independent factors affecting the ECV success rate, and we scored them using a regression coefficient. RESULTS: Women with scores of 0-3 points had ECV success rates, vaginal delivery rates, and delivery gestational ages at 16.67%, 16.67%, and 38.88 weeks, respectively; those with scores of 4-6 points had ECV success rates, vaginal delivery rates, and delivery gestational ages at 65.75%, 58.90%, and 39.62 weeks, respectively; and those with scores of 7-9 points had ECV success rates, vaginal delivery rates, and delivery gestational ages at 93.71%,74.83%, and 40.00 weeks, respectively. CONCLUSIONS: The ECV success and vaginal delivery rates increased with the score, and the delivery gestational age showed an initial increase. To optimize the ECV procedure and reduce the hospital burden, this scoring system should be used routinely to predict the ECV success rate and determine the timing of ECV attempts.

Isolation of a Bacillus cereus strain HBL-AI and its application for production of Trans-4-hydroxy-l-proline.

A new trans-4-hydroxy-l-proline (trans-Hyp) producing Bacillus cereus HBL-AI, was isolated from the air, which was screened just using l-proline as carbon and energy sources. This strain exhibited 73·4% bioconversion rate from initial l-proline (3 g l-1 ) to trans-Hyp. By sequencing the genome of this bacterium, 6244 coding sequences were obtained. Genome annotation analysis and functional expression were used to identify the proline-4-hydroxylase (BP4H) in HBL-AI. This enzyme belonged to a family of 2-oxoglutarate-related dioxygenases, which required 2-oxoglutarate and O2 as co-substrates for the reaction. Homologous modelling indicated that the enzyme had two monomers and contained conserved motifs, which included a distorted 'jelly roll' ß strand core and the residues (HXDXnH and RXS). The engineering Escherichia coli 3 Δ W3110/pTrc99a-proba-bp4h was constructed using BP4H, which transformed glucose to trans-Hyp in one step with high concentration of 46·2 g l-1 . This strategy provides a green and efficient method for synthesis of trans-Hyp and thus has a great potential in industrial application.

LncRNA XIXT promotes osteogenic differentiation of bone mesenchymal stem cells and alleviates osteoporosis progression by targeting miRNA-30a-5p.

OBJECTIVE: To uncover the role of XIXT in influencing the osteogenesis of hBMSCs by adsorbing microRNA-30a-5p (miRNA-30a-5p) to upregulate RUNX2. PATIENTS AND METHODS: The serum samples were collected from osteoporosis patients and normal people. hBMSCs were isolated from femoral head tissues. The serum levels of XIXT and miRNA-30a-5p were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The expression levels and activities of the osteogenic differentiation-related genes in hBMSCs after transfection of sh-XIXT, sh-RUNX2, miRNA-30a-5p mimic, and inhibitor were detected by qRT-PCR, Western blot, ALP activity assay, and alizarin red staining. The Dual-Luciferase Reporter Gene Assay was performed to confirm the binding of XIXT to miRNA-30a-5p, as well as the binding of miRNA-30a-5p to RUNX2. RESULTS: LncRNA XIXT was significantly downregulated, and miRNA-30a-5p was upregulated in the serum of osteoporosis patients. The osteogenic differentiation-related genes (ALP, RUNX2) and XIXT were markedly upregulated in a time-dependent manner, while miRNA-30a-5p level gradually decreased in hBMSCs with the prolongation of osteogenesis. The knockdown of XIXT inhibited the osteogenic differentiation of hBMSCs. In hBMSCs, XIXT regulated RUNX2 expression by targeting miRNA-30a-5p. The knockdown of miRNA-30a-5p partially reversed the inhibitory effect of XIXT on the osteogenesis of hBMSCs. However, the downregulated RUNX2 reversed the promotive effect of miRNA-30a-5p on the osteogenesis of hBMSCs. CONCLUSIONS: LncRNA XIXT upregulated RUNX2 by absorbing miRNA-30a-5p, and thus induced hBMSCs osteogenesis to alleviate osteoporosis.

MiR-302 a/b/c suppresses tumor angiogenesis in hepatocellular carcinoma by targeting MACC1.

OBJECTIVE: Hepatocellular carcinoma (HCC) is a hypervascularized tumor. Aberrant angiogenesis is the main cause, which results in cancer growth and progression. It has been showed that microRNA-302 cluster (miR-302) may be associated with angiogenesis. Here, we aimed to identify the role of miR-302a/b/c in the regulation of cell angiogenesis in HCC. PATIENTS AND METHODS: MRNA expression of miR-302a/b/c and MACC1 was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The protein of MACC1 was measured using Western blot. Cells proliferation, migration, and invasion abilities were investigated via Cell Counting Kit-8 (CCK-8) assay or transwell assay, respectively. Tube formatting assays were used to explore the tube formation capacity. The interaction among miR-302a/b/c was analyzed by luciferase assay. RESULTS: The expression of miR-302a/b/c was greatly reduced while MACC1 expression, whether mRNA or protein was conspicuously elevated in HCC tissues and cells. Then, functional experiment results showed miR-302a/b/c overexpression and MACC1 down-regulation inhibited the proliferation, migration, invasion ability, and tube formation capacity of HUVECs. In addition, we detected that miR-302a/b/c directly targeted MACC1 and suppressed MACC1 expression, and miR-302a/b/c could suppress tumor angiogenesis in HCC by targeting MACC1. CONCLUSIONS: MiR-302a/b/c may function as a potential suppressor of tumor angiogenesis in HCC by targeting MACC1, indicating a promising target for HCC therapy.

Correlations of the MiR-330 expression with the pathogenesis and prognosis of breast cancer.

OBJECTIVE: To investigate the expression of micro ribonucleic acid-330 (miR-330) in breast cancer tissues and cancer-adjacent tissues as well as the correlations of the miR-330 expression with clinicopathological features and the prognosis of breast cancer patients. PATIENTS AND METHODS: The expression levels of miR-330 in cancer tissues and cancer-adjacent tissues of 120 breast cancer patients with complete follow-up data were detected via the reverse transcription-polymerase chain reaction (RT-PCR). Meanwhile, the expression level of miR-330 in serum of breast cancer patients was measured using enzyme-linked immunosorbent assay (ELISA). The correlations of the expression level of miR-330 with clinicopathological data and the prognosis of breast cancer patients were explored. RESULTS: The expression level of miR-330 in breast cancer tissues was remarkably higher than that in cancer-adjacent tissues, and it was also higher in serum of breast cancer patients than that of healthy controls. In breast cancer patients with axillary lymph node metastasis, the proportion of patients with the high expression of miR-330 [30/54 (55.6%)] was markedly larger than that of patients with the low expression of miR-330 [6/30 (20%)] (p=0.003). In terms of tumor-node-metastasis (TNM) stage, the proportion of patients with the high expression of miR-330 in stage II or above was evidently larger than that of patients with the low expression of miR-330, in which the proportion was 46/60 (77.2%) in stage III and 11/15 (73.3%) in stage IV (p=0.002). Additionally, the tumor size, the histological grade, the expression of human epidermal growth factor receptor 2 (HER2), the expression of hormone receptors and the tissue type, were not related to the expression level of miR-330 (p>0.05). It was manifested in the survival curve that the median survival time was 84.8 months in the miR-330 high expression group and 96.8 months in the miR-330 low expression group, displaying a statistical difference (p=0.01). CONCLUSIONS: MiR-330 is highly expressed in cancer tissues and serum of patients with breast cancer, and it can promote the axillary lymph node metastasis, which is an important factor affecting the prognosis of breast cancer patients. However, no obvious correlations of the expression level of miR-330 with the tumor size, the histological grade, the HER2 expression and the expression of estrogen receptors are found.

Enantioselective synthesis of enantiopure chiral alcohols using carbonyl reductases screened from Yarrowia lipolytica.

AIMS: We aimed to explore Yarrowia lipolytica carbonyl reductases as effective biocatalysts and to develop efficient asymmetric reduction systems for chiral alcohol synthesis. METHODS AND RESULTS: Yarrowia lipolytica carbonyl reductase genes were obtained via homologous sequence amplification strategy. Two carbonyl reductases, YaCRI and YaCRII, were identified and characterized, and used to catalyse the conversion of 2-hydroxyacetophenone (2-HAP) to optically pure (S)-1-phenyl-1,2-ethanediol. Enzymatic assays revealed that YaCRI and YaCRII exhibited specific activities of 6·96 U mg-1 (99·8% e.e.) and 7·85 U mg-1 (99·9% e.e.), respectively, and showed moderate heat resistance at 40-50°C and acid tolerance at pH 5·0-6·0. An efficient whole-cell two-phase system was established using reductase-expressing recombinant Escherichia coli. The conversion of 2-HAP (20·0 g l-1 ) conversion with the solvent of dibutyl phthalate was approximately 70-fold higher than in water. Furthermore, the two recombinant E. coli displayed biocatalyst activity and enantioselectivity towards several different carbonyl compounds, and E. coli BL21 (DE3)/pET-28a-yacrII showed a broad substrate spectrum. CONCLUSIONS: A new whole-cell recombinant E. coli-based bioreduction system for enantiopure alcohol synthesis with high enantioselectivity at high substrate concentrations was developed. SIGNIFICANCE AND IMPACT OF THE STUDY: We proposed a promising approach for the efficient preparation of enantiopure chiral alcohols.

Endophytic Bacillus strains isolated from alfalfa (Medicago sativa L.) seeds: enhancing the lifespan of Caenorhabditis elegans.

Medicago sativa L. (alfalfa) is an important forage crop throughout the world. Despite the abiotic nutritional components of alfalfa having been extensively studied, there is only limited information on alfalfa endophytes. In this study, thirteen endophytic bacteria were isolated from alfalfa seeds. Bacillus (76·9%) was the most abundant genus, followed by Enterobacter (15·4%), Brevibacterium (7·7%), Geobacillus (7·7%) and Staphylococcus (7·7%). Four of the 13 endophytic bacteria, including Bacillus amyloliquefaciens EnB-alf1, Bacillus subtilis EnB-alf3, EnB-alf5 and EnB-alf13, were capable of significantly extending the lifespan of Caenorhabditis elegans. In addition, B. amyloliquefaciens EnB-alf1 enhanced the resistance of C. elegans to thermal stress whereas B. subtilis EnB-alf3 enhanced the resistance to oxidative stress. Further studies demonstrated that the enhanced lifespan of the worm was depended on the function of DAF-2/DAF-16 and was associated with the colonization of strain in the worms' intestines when strain EnB-alf1 or strain EnB-alf3 was presented to the worms as food sources. Our results suggest that some endophytic Bacillus strains isolated from alfalfa are beneficial on C. elegans health. SIGNIFICANCE AND IMPACT OF THE STUDY: Medicago sativa L. (alfalfa) is an important forage crop throughout the world. Despite the abiotic nutritional components of alfalfa having been extensively studied, there is only limited information available on alfalfa endophytes. Beneficial bacteria residing in the host intestine have been shown to affect host longevity. However, there is limited information available on the functions of alfalfa seed endophytes to nematodes. In this study, four endophytic Bacillus strains isolated from alfalfa seeds were found to significantly extend the lifespan of Caenorhabditis elegans and enhance resistance to thermal and oxidative stress. Our results suggest that some endophytic Bacillus strains isolated from alfalfa seeds can promote good health in C. elegans.

The effect of miR-146a gene silencing on drug-resistance and expression of protein of P-gp and MRP1 in epilepsy.

OBJECTIVE: To investigate the effect of miR-146a gene silencing on brain tissue and related drug-resistance proteins in rats and explore its resistance mechanism. MATERIALS AND METHODS: A rat model of chronic refractory epilepsy was established. The rats were divided into four groups: Normal group, Model group, Negative control group and AntagomiR-146a group. Hematoxylin and eosin (HE) stain was used to detect brain histopathological changes. We examined the expression of mRNA of miR-146a, multidrug resistance (MDR1) and multidrug-resistant associated protein (MRP1) by RT-PCR. The expressions of protein of High motility group box 1 (HMGB1)/Toll-like receptor 4 (TLR4)/nuclear transcription factor-κB (NF-κB) pathway and P-glycoprotein (P-gp), MRP1 were detected by Western-blotting. RESULTS: We demonstrated that the pathological lesion was lighter in antagomiR-146a group compared with the model group. The mRNA expression of miR-146a in AntagomiR-146a group was significantly decreased compared to the model group. Furthermore, the mRNA expression of MDR1 and MRP1 in AntagomiR-146a group was lower than that in the model group. In addition, the protein expression of HMGB1, TLR4, NF-κB and P-gp, MRP1 in AntagomiR-146a group was lower than that in model group. CONCLUSIONS: These results demonstrated that miR-146a gene silencing can attenuate pathological changes and improve drug resistance in refractory epilepsy. Also, it is closely related to the HMGB1/TLR4/NF-κB signaling pathway regulation.

Expression of miR-146a-5p in patients with intracranial aneurysms and its association with prognosis.

OBJECTIVE: The study aims to detect the association of miR-146a-5p with intracranial aneurysms (IAs). PATIENTS AND METHODS: The expression of miR-146a-5p was compared from plasma samples between 72 patients with intracranial aneurysms (IAs) and 40 healthy volunteers by quantitative Real-time polymerase chain reaction (qRT-PCR). Statistical analysis was performed to analyze the relationship between miR-146a-5p expression and clinical data and overall survival (OS) time of IAs patients. Univariate and multivariate Cox proportional hazards have also been performed. RESULTS: Notably, higher miR-146a-5p expression was found in plasma samples from 72 patients with intracranial aneurysms (IAs) compared with 40 healthy controls. Higher miR-146a-5p expression was significantly associated with rupture and Hunt-Hess level in IAs patients. Kaplan-Meier survival analysis verified that higher miR-146a-5p expression predicted a shorter overall survival (OS) compared with lower miR-146a-5p expression in IAs patients. Univariate and multivariate Cox proportional hazards demonstrated that higher miR-146a-5p expression, rupture, and Hunt-Hess were independent risk factors of OS in patients with intracranial aneurysms (IAs). CONCLUSIONS: MiR-146a-5p expression may serve as a biomarker for predicting prognosis in patients with IAs.

Effect of TAGLN2 in the regulation of meningioma tumorigenesis and development.

OBJECTIVE: To explore the role of transgelin-2 TAGLN2 in the development and progression of meningioma and the potential regulatory. MATERIALS AND METHODS: TAGLN2 knockdown expression and overexpression in vitro models were constructed using lentivirus in meningioma cell line CH157; their corresponding transfection efficiencies were verified by qRT-PCR and Western Blot. Actions of TAGLN2 on the proliferation of meningioma cells were explored by CCK8 and colony formation assays. The effect of TAGLN2 on invasion of meningioma cells was analyzed by transwell cell invasion assay. Biological function of TAGLN2 on apoptosis of meningioma cells was determined by flow cytometry. Finally, Western Blot was used to investigate the detailed mechanism of TAGLN2 on regulating the biological functions of meningioma cells. RESULTS: After down-regulating the expression of TAGLN2, there were significantly decreased capacities of cells proliferation and colony formation of meningioma cells, meanwhile, cell invasion was significantly decreased but the apoptosis rate was increased. On the contrary, up-regulation of TAGLN2 expression, the proliferation, colony formation ability were significantly increased as well as the invasion capacity, whilst apoptosis rate was decreased. Western Blot showed that expressions of p-PI3K and p-AKT were inhibited after knockdown of TAGLN2, which were significantly increased after TAGLN2 was overexpressed. CONCLUSIONS: TAGLN2 can affect the proliferation, invasion and apoptosis of meningioma cells and may participate in the development of meningioma through regulating the PI3K/AKT signaling pathway.

Efficient production of trans-4-Hydroxy-l-proline from glucose by metabolic engineering of recombinant Escherichia coli.

Trans-4-Hydroxy-l-proline (trans-Hyp) is a valuable chiral building block for the synthesis of pharmaceutical intermediates. Bioconversion of l-proline using recombinant strain with proline-4-hydroxylase (P4H) is a preferred biocatalytic process in the economical production of trans-Hyp. In this study, a recombinant E. coli overexpressing hydroxylase (P4H), γ-glutamyl kinase and glutamate-semialdehyde dehydrogenase (ProBA) genes were constructed by knocking out the key genes in the metabolism. These key genes contained putA encoding proline dehydrogenase (PutA) in the l-proline metabolism and other catalytic enzyme genes, sucAB encoding α-ketoglutarate dehydrogenase (SucAB), aceAK encoding isocitratelyase (AceA) and isocitrate dehydrogenase kinase/phosphatase (AceK) in the TCA cycle. This recombinant strain coupled the synthetic pathway of trans-Hyp with TCA cycle of the host strain. It inhibited the consumption of l-proline completely and promoted the accumulation of 2-oxoglutarate (2-OG) as a co-substrate, which realized the highest conversion of glucose to trans-Hyp. A fed-batch strategy was designed, capable of producing 31·0 g l-1 trans-Hyp from glucose. It provided a theoretical basis for commercial conversion of glucose to trans-Hyp. SIGNIFICANCE AND IMPACT OF THE STUDY: Trans-4-Hydroxy-l-proline (trans-Hyp) is a valuable chiral building block for the synthesis of pharmaceutical intermediates. Unsatisfactory microbial bioconversion resulted in a low yield of trans-Hyp. In this study, we blocked the unwanted blunting pathways of host strain and make the cell growth couple with the trans-Hyp synthesis from glucose. Finally, a recombinant Escherichia coli with short-cut and efficient trans-Hyp biosynthetic pathway was obtained. It provided a theoretical basis for commercial production of trans-Hyp.

Arachidonic acid attenuates brain damage in a rat model of ischemia/reperfusion by inhibiting inflammatory response and oxidative stress.

The aim of the present study was to study the effects of arachidonic acid (ARA) in a rat brain ischemia/reperfusion model induced by middle cerebral artery occlusion (MCAO). A total of 50 rats were randomly divided into five groups: control group, MCAO group, MCAO + ARA 0.3 g/kg group, MCAO + ARA 1 g/kg group, and MCAO + ARA 3 g/kg group. The MCAO + ARA groups received ARA by intraperitoneal injection daily for 14 consecutive days, while the rats in the control and MCAO groups were given equivalent volume of saline. We detected the Morris water maze test and pathological changes to investigate the ischemia/reperfusion injury. The protein levels of tumor necrosis factor-alpha and interleukin-6 in the hippocampus were detected by enzyme-linked immunosorbent assay kits. In addition, the activities of superoxide dismutase, glutathione peroxidase, and malondialdehyde were assayed in hippocampus homogenates to evaluate the oxidative stress after ischemia/reperfusion. The results indicated that ARA administration decreased biochemical parameters of inflammation and oxidative stress. Morris water maze test and histopathological examination further verified the protective effects of ARA on ischemia/reperfusion injury rats. These findings demonstrated that ARA could protect MCAO-induced brain injury rats by inhibition of inflammation and oxidative stress, suggesting that it may have potential as a therapy for cerebral ischemia/reperfusion injury.

A cross-sectional study of acute diarrhea in Pudong, Shanghai, China: prevalence, risk factors, and healthcare-seeking practices.

Diarrhea is a common cause of morbidity and mortality and the incidence of diarrhea in the world has changed little over the past four decades. To assess the prevalence of and healthcare practices for diarrhea, a cross-sectional study was conducted in Pudong, Shanghai, China. In October 2014, a total of 5324 community residents were interviewed. Respondents were asked if they had experienced diarrhea (defined as ⩾3 passages of watery, loose, bloody, or mucoid stools within a 24-h period) in the previous month prior to the interview. The monthly prevalence of diarrhea was 4·1% (95% CI: 3·3-4·8), corresponding to an incidence rate of 0·54 episodes per person-year. The proportion of individuals with diarrhea who sought healthcare was 21·2% (95% CI: 13·4-29·0). Diarrhea continues to impose a considerable burden on the community and healthcare system in Pudong. Young age and travel were identified as predictors of increased diarrhea occurrence.

MiR-195 enhances cardiomyocyte apoptosis induced by hypoxia/reoxygenation injury via downregulating c-myb.

OBJECTIVE: In this study, we explored the regulative effect of miR-195 on c-myb expression and also investigated the role of miR-195 and c-myb in cardiomyocyte apoptosis induced by hypoxia/reoxygenation (H/R) injury. MATERIALS AND METHODS: QRT-PCR analysis was performed to measure mature miR-195 expression. H9c2 cells were transfected for miR-195 overexpression or knockdown or c-myb overexpression using Lipofectamine 2000. The cells were subjected to H/R treatment and following flow cytometric analysis of active caspase-3 or florescent study of reactive oxygen species (ROS) generation. The binding sites between miR-195 and 3'UTR of MYB mRNA were predicted using TargetScan 7.0. The binding sites were verified using dual luciferase assay and Western blot analysis. RESULTS: MiR-195 is significantly upregulated after H/R treatment in H9c2 cells. H/R injury induced active caspase-3 expression. However, the cells with miR-195 suppression had substantially lower ratio of cells with active caspase-3. MiR-195 can decrease c-myb protein expression. Dual luciferase assay verified two binding sites between miR-195 and 3'UTR of MYB mRNA. C-myb overexpression can suppress mitochondrial superoxide generation and cardiomyocyte apoptosis after H/R. CONCLUSIONS: MiR-195 is significantly increased due to H/R and can enhance cardiomyocyte apoptosis. MYB is a target gene of miR-195 in cardiomyocytes. The miR-195-MYB axis is involved in regulation of cardiomyocyte apoptosis induced by H/R.