Efficient generation of functional hepatocyte-like cells from mouse liver progenitor cells via indirect co-culture with immortalized human hepatic stellate cells.

BACKGROUND: Differentiation of liver progenitor cells (LPCs) to functional hepatocytes holds great potential to develop new strategies for hepatocyte transplantation and the screening of drug-induced cytotoxicity. However, reports on the efficient and convenient hepatic differentiation of LPCs to hepatocytes are few. The present study aims to investigate the possibility of generating functional hepatocytes from LPCs in an indirect co-culture system. METHODS: Mouse LPCs were co-cultured in Transwell plates with an immortalized human hepatic stellate cell line (HSC-Li) we previously established. The morphology, expression of hepatic markers, and functions of mouse LPC-derived cells were monitored and compared with those of conventionally cultured LPCs. RESULTS: Co-culturing with HSC-Li cells induced differentiation of mouse LPCs into functional hepatocyte-like cells. The differentiated cells were morphologically transformed into hepatocyte-like cells 3 days after co-culture initiation. In addition, the differentiated cells expressed liver-specific genes and possessed hepatic functions, including glycogen storage, low-density lipoprotein uptake, albumin secretion, urea synthesis, and cytochrome P450 1A2 enzymatic activity. CONCLUSIONS: Our method, which employs indirect co-culture with HSC-Li cells, can efficiently induce the differentiation of LPCs into functional hepatocytes. This finding suggests that this co-culture system can be a useful method for the efficient generation of functional hepatocytes from LPCs.

Inverse relationship between serum osteocalcin levels and nonalcoholic fatty liver disease in postmenopausal Chinese women with normal blood glucose levels.

AIM: Osteocalcin is involved in the progression of nonalcoholic fatty liver disease (NAFLD) in animal models and humans. In this study we investigated the relationship between serum osteocalcin levels and NAFLD in postmenopausal Chinese women. METHODS: A total of 733 postmenopausal women (age range: 41-78 years) with normal blood glucose levels were enrolled in this cross-sectional study. Women taking lipid-lowering or anti-hypertensive drugs were excluded. Serum osteocalcin levels were assessed using an electrochemiluminescence immunoassay. The degree of NAFLD progression for each subject was assessed through ultrasonography. The fatty liver index (FLI) of each subject was calculated to quantify the degree of liver steatosis. RESULTS: The median level of serum osteocalcin for all subjects enrolled was 21.99 ng/mL (interquartile range: 17.84-26.55 ng/mL). Subjects with NAFLD had significantly lower serum osteocalcin levels (18.39 ng/mL; range: 16.03-23.64 ng/mL) compared with those without NAFLD (22.31 ng/mL; range: 18.55-27.06 ng/mL; P<0.01). Serum osteocalcin levels decreased with incremental changes in the FLI value divided by the quartile (P-value for trend<0.01). The serum osteocalcin levels showed a negative correlation with the FLI values, even after adjusting for confounding factors (standardized ß=-0.124; P<0.01). Binary logistic regression analysis identified an individual's serum osteocalcin level as an independent risk factor for NAFLD (odds ratio: 0.951; 95% confidence interval: 0.911-0.992; P=0.02). CONCLUSION: Serum osteocalcin levels are inversely correlated with NAFLD in postmenopausal Chinese women with normal blood glucose levels.

Predictors of early post ischemic stroke apathy and depression: a cross-sectional study.

BACKGROUND: Apathy and depression are important neuropsychiatric disorders that can occur after a stroke but the etiology and risk factors are not well understood. The purpose of this study was to identify risk factors for apathy and depression following a stroke. METHODS: Patients with an acute stroke who met the inclusion criteria were recruited from our hospital, and general information was recorded from patient charts. The Apathy Evaluation Scale, Clinician Version (AES-C) was used to evaluate these patients within 2 weeks after the stroke. The Montreal Cognitive Assessment (MoCA), mini-mental state examination (MMSE), Hamilton Depression Scale (HAMD), Mattis Dementia Rating Scale Initiation/Perseveration subset (MDRS I/P), Frontal Assessment Battery (FAB) and Stroop Color-Word Association Test were employed to evaluate emotion, cognitive function and executive function. The patients were divided into two groups: the apathy group and the non-apathy group. We also divided the patients into two groups based on whether or not they had post-stroke depression. The clinical characteristics and scores on the MoCA, MMSE, HAMD and MDRS I/P were compared between the apathy and non-apathy groups as well as between patients with and without depression. Logistic regression analysis was performed to identify risk factors for apathy and depression following a stroke. RESULTS: A total of 75 patients with acute stroke were recruited. Of these, 25 (33.3%) developed apathy and 12 (16%) developed depression. Multivariate logistic regression analysis indicated that a history of cerebrovascular disease (OR: 6.45, 95% CI: 1.48-28.05, P = 0.013), low HbA1c (OR: 0.31, 95% CI: 0.12-0.81, P = 0.017) and a low MDRS I/P score (OR: 0.84, 95% CI: 0.74, 0.96, P = 0.010) were risk factors for post-stroke apathy. Additionally, multivariate logistic regression indicated that a low MDRS I/P (OR: 0.85, 95% CI: 0.75, 0.97, P = 0.015) was associated with post-stroke depression. CONCLUSIONS: Three risk factors for post-stroke apathy were identified as a history of cerebrovascular disease, low HbA1c and lower MDRS I/P scores. A low MDRS I/P score was also identified as a risk factor for post-stroke depression. These results may be useful to clinicians in recognizing and treating apathy and depression in patients after a stroke.

Physcion increases the sensitivity of hepatocellular carcinoma to sorafenib through miRNA-370/PIM1 axis-regulated glycolysis.

BACKGROUND: Resistance to sorafenib has become a challenge in clinical treatment of hepatocellular carcinoma (HCC). Physcion is a common bioactive anthraquinone that has potential as an anticancer agent. AIM: To study the effect of physcion on sensitizing HCC cells to sorafenib. METHODS: Sorafenib-resistant HCC cells were established and treated with sorafenib and/or physcion. The cell viability, proliferation and apoptosis were measured by cell counting kit-8, colony formation, flow cytometry, and in vivo xenograft model. Glucose uptake, lactate acid production, extracellular acidification rate (ECAR), and oxygen consumption rate (OCR) were measured to analyze glycolysis. Expression of glycolysis-related regulators was assessed by western blotting. RESULTS: The addition of physcion significantly enhanced the antitumor effects of sorafenib on sorafenib-resistant HCC cells, manifested by enhanced apoptosis and suppressed cell growth. The glucose uptake, lactate acid production, and ECAR were elevated, and OCR was suppressed by physcion treatment. The level of PIM1 was elevated and miR-370 was suppressed in sorafenib-resistant HCC cells compared with the parental cells, which was suppressed by physcion treatment. Inhibition of miR-370 notably reversed the effects of physcion on sorafenib-resistant HCC cells. CONCLUSION: Our data indicated that physcion enhanced the sensitivity of HCC cells to sorafenib by enhancing miR-370 to suppress PIM1-promoted glycolysis.

Benefits of Mindfulness Training on the Mental Health of Women During Pregnancy and Early Motherhood: A Randomized Controlled Trial.

Objective: This study aimed to evaluate the effects of a mindfulness-based psychosomatic intervention on depression, anxiety, fear of childbirth (FOC), and life satisfaction of pregnant women in China. Methods: Women experiencing first-time pregnancy ( n = 104) were randomly allocated to the intervention group or a parallel active control group. We collected data at baseline (T0), post-intervention (T1), 3 days after delivery (T2), and 42 days after delivery (T3). The participants completed questionnaires for the assessment of the levels of depression, anxiety, FOC, life satisfaction, and mindfulness. Differences between the two groups and changes within the same group were analyzed at four time points using repeated-measures analysis of variance. Results: Compared with the active control group, the intervention group reported lower depression levels at T2 ( P = 0.038) and T3 ( P = 0.013); reduced anxiety at T1 ( P = 0.001) and T2 ( P = 0.003); reduced FOC at T1 ( P < 0.001) and T2 ( P = 0.04); increased life satisfaction at T1 ( P < 0.001) and T3 ( P = 0.015); and increased mindfulness at T1 ( P = 0.01) and T2 ( P = 0.006). Conclusion: The mindfulness-based psychosomatic intervention effectively increased life satisfaction and reduced perinatal depression, anxiety, and FOC.

Advances in cell sources of hepatocytes for bioartificial liver.

BACKGROUND: Orthotopic liver transplantation (OLT) is the most effective therapy for liver failure. However, OLT is severely limited by the shortage of liver donors. Bioartificial liver (BAL) shows great potential as an alternative therapy for liver failure. In recent years, progress has been made in BAL regarding genetically engineered cell lines, immortalized human hepatocytes, methods for preserving the phenotype of primary human hepatocytes, and other functional hepatocytes derived from stem cells. DATA SOURCES: A systematic search of PubMed and ISI Web of Science was performed to identify relevant studies in English language literature using the key words such as liver failure, bioartificial liver, hepatocyte, stem cells, differentiation, and immortalization. More than 200 articles related to the cell sources of hepatocyte in BAL were systematically reviewed. RESULTS: Methods for preserving the phenotype of primary human hepatocytes have been successfully developed. Many genetically engineered cell lines and immortalized human hepatocytes have also been established. Among these cell lines, the incorporation of BAL with GS-HepG2 cells or alginate-encapsulated HepG2 cells could prolong the survival time and improve pathophysiological parameters in an animal model of liver failure. The cBAL111 cells were evaluated using the AMC-BAL bioreactor, which could eliminate ammonia and lidocaine, and produce albumin. Importantly, BAL loading with HepLi-4 cells could significantly improve the blood biochemical parameters, and prolong the survival time in pigs with liver failure. Other functional hepatocytes differentiated from stem cells, such as human liver progenitor cells, have been successfully achieved. CONCLUSIONS: Aside from genetically modified liver cell lines and immortalized human hepatocytes, other functional hepatocytes derived from stem cells show great potential as cell sources for BAL. BAL with safe and effective liver cells may be achieved for clinical liver failure in the near future.

Key challenges to the development of extracorporeal bioartificial liver support systems.

BACKGROUND: For nearly three decades, extracorporeal bioartificial liver (BAL) support systems have been anticipated as promising tools for the treatment of liver failure. However, these systems are still far from clinical application. This review aimed to analyze the key challenges to the development of BALs. DATA SOURCE: We carried out a PubMed search of English-language articles relevant to extracorporeal BAL support systems and liver failure. RESULTS: Extracorporeal BALs face a series of challenges. First, an appropriate cell source for BAL is not readily available. Second, existing bioreactors do not provide in vivo-like oxygenation and bile secretion. Third, emergency needs cannot be met by current BALs. Finally, the effectiveness of BALs, either in animals or in patients, has been difficult to document. CONCLUSIONS: Extracorporeal BAL support systems are mainly challenged by incompetent cell sources and flawed bioreactors. To advance this technology, future research is needed to provide more insights into interpreting the conditions for hepatocyte differentiation and liver microstructure formation.

[Metabolic syndrome and microalbuminuria accompanied with hyperglycemia in community subjects].

OBJECTIVE: To examine the prevalence of metabolic syndrome (MS) and microalbuminuria (MAU) in subjects with different levels of glucose tolerance and probe the risk factors for the development of MAU. METHODS: A total of 951 subjects were divided into 3 groups according to the results of oral glucose tolerance test (OGTT). Among them, there were 674 subjects with normal glucose tolerance (NGT), 195 with impaired glucose regulation (IGR) and 82 newly-diagnosed cases of type 2 diabetes mellitus (T2DM). MAU was diagnosed if urine albumin-to-creatinine ratio (UACR) was 30 - 300 mg/g. RESULTS: (1) Compared to the NGT subjects, both the IGR and newly-diagnosed T2DM subjects had the significantly higher levels of age, body mass index (BMI), waist circumference, fasting plasma glucose, 2-hour post OGTT glucose (2 hPG), glycated hemoglobin, glycated albumin, fasting insulin, homeostasis model of assessment-insulin resistance (HOMA-IR), triglyceride, systolic blood pressure (SBP) and UACR but a lower level of high density lipoprotein-cholesterol (HDL-C); (2) The incident rate of MS and MAU in the IGR and newly-diagnosed T2DM subjects was 54.4% (106/195), 12.3% (24/195) and 61.0% (50/82), 12.2% (10/82) respectively versus 9.1% (61/674) and 4.9% (33/674) in the NGT subjects. The incident rate of abdominal obesity, hypertension, hypertriglyceridemia, low HDL-C, MS and MAU in the IGR and newly-diagnosed T2DM subjects was significantly higher than that of the NGT subjects (all P < 0.05); (3) Multiple regression analysis showed that BMI, SBP and 2 hPG were independently associated with UACR. Logistic regression analysis indicated that BMI, SBP and 2 hPG were independent risk factors of MAU. CONCLUSIONS: The prevalence of MS and MAU is significantly higher in the IGR and newly-diagnosed T2DM subjects. BMI, SBP and 2 hPG are independent risk factors of MAU.

Cost-effectiveness analysis of the prevention of mother-to-child transmission of HIV.

BACKGROUND: The number of HIV-positive pregnant women accounted for about 10% of China's total over the past few years in Liangshan Prefecture, Sichuan province in China. Although cost-effectiveness of the PMTCT of HIV have been evaluated in other previous studies, no specific study has been conducted in Liangshan prefecture, nor has the expenses paid individually by HIV-positive pregnant women been included. The purpose of this study was to evaluate both the short-term and long-term cost-effectiveness of PMTCT of HIV in Liangshan Prefecture from the social perspective. METHODS: From December 2018 to January 2019, individual expenses and the other costs were collected: individual expenses of 133 recruited HIV-positive pregnant women registered in the National Information System of Prevention of Mother-to-Child Transmission of HIV, Syphilis, and HBV, and the other costs from local maternal and child healthcare hospitals, Centers for Disease Control and Prevention, and general hospitals. The costs, the number of pediatric infections averted from being HIV infected were analyzed. And, Life years gained by pediatric infections averted were calculated by using a life table. Besides, Direct benefit was calculated through a Markov mode. Furthermore, One-way sensitivity analysis was conducted for key variables affecting the benefit-cost ratio. RESULTS: The estimated number of pediatric infections averted was 164.The total cost was USD 114.1 million, including direct medical costs, direct non-medical costs, and indirect costs, which were USD 54.2 million, USD 53.4 million, and USD 6.5 million, respectively. 630.6 person-years discounted to 2017 were gained at a 3% annual rate, and cost per life year gained was USD 1809.50. Direct benefits were USD 198.4 million, indirect benefits USD 82.5 million, and the benefit-cost ratio was 1.5. The sensitivity analysis showed that if PMTCT costs hypothetically ranged from USD 85.6 million to USD 142.6 million, benefit-cost ratio would vary from 1.0 to 2.3. CONCLUSIONS: PMTCT of HIV in Liangshan Prefecture was very cost-effective. It was a great economic burden of PMTCT on HIV-positive pregnant women and their families to take individual expenses. Therefore, it could be suggested that individual expenses should be covered as much as possible by different types of financing.

Understanding the Choice of Sleep Arrangements and Soothing Methods and Their Associations with Sleep Problems among Children Under 3 Years Old: A Chinese Population-Based Study.

Objective: This study aimed to examine the sleep arrangements and soothing methods and to assess their associations with sleep problems among children aged < 3 years in China. Methods: A cross-sectional survey was conducted in 2019 from six provinces in China. A total of 1,195 caregivers of children aged 0-35 months were included in the study. Data on sleep arrangements, soothing methods, and sleep problems (i.e., frequent night awakenings and difficulty falling asleep) were assessed using the Brief Infant Sleep Questionnaire. The reasons for bed-sharing in sleep arrangements were recorded using a self-designed questionnaire. Results: The bed-sharing practice was very prevalent at any age, which ranged from 69.9% to 78.3%. Most infants fell asleep while feeding or being rocked/held before age 12 months. By age 35 months, 62.4% of the children fell asleep in bed near parents. The most common reasons for bed-sharing were breastfeeding/feeding and convenience. Parental involvement when falling asleep was significantly related with frequent night awakenings and difficulty falling asleep. No association was found between bed-sharing and sleep. Conclusion: Bed-sharing and parental involvement were very common among Chinese children aged < 3 years. Children who fall asleep with parental involvement were more likely to have sleep problems.

Relationship between serum osteocalcin and glycaemic variability in Type 2 diabetes.

1. Recent reports have described the role of osteocalcin in glucose metabolism and glycaemic variability has been proven to be associated with an increased risk of diabetes complications. However, the relationship between osteocalcin and glycaemic variability remains unclear. The aim of the present study was to examine the relationship between serum osteocalcin and glycaemic variability, as determined by a continuous glucose monitoring (CGM) system in patients with Type 2 diabetes mellitus (T2DM). 2. Fifty-nine T2DM patients with glycosylated haemoglobin (HbA1c) levels between 7.0% and 10.9% were recruited to the present study. Biochemical information and CGM parameters were collected at baseline and after 8 weeks of antihyperglycaemic therapy (either sulphonylurea, sulphonylurea + an α-glucosidase inhibitor or insulin + metformin combination therapy). 3. Compared with baseline, serum osteocalcin increased significantly (P = 0.014), whereas parameters related to glucose variability, including the mean amplitude of glycaemic excursions (MAGE) and the standard deviation of blood glucose values, decreased significantly (P < 0.001) after the 8 week treatment period. At baseline, there was a positive correlation between serum osteocalcin levels and fasting C-peptide levels (P = 0.004) and homeostatic model assessment of ß-cell function (P = 0.048), but a negative correlation between serum osteocalcin levels and fasting plasma glucose (P = 0.023), HbA1c (P = 0.020), glycated albumin (P = 0.019) and 24 h mean blood glucose (P < 0.001). Multiple stepwise regression analysis indicated that baseline osteocalcin was the single parameter that best predicted the change in MAGE (ß = -0.122; P = 0.039). 4. In conclusion, serum osteocalcin concentrations increased with improved glucose control. High initial osteocalcin levels were associated with subsequent improvements in glucose variability during glucose-lowering treatment.

Investigation Regarding Early Cognitive Function of Women in the Postpartum Period and the Analysis of Influencing Factors.

OBJECTIVE: The present study aims to assess the cognitive function of healthy full-term puerperae and compare it with the cognitive function of healthy non-pregnant women in order to analyze possible influencing factors. METHODS: The study subjects were divided into two groups: the maternal (case) group (n = 80) and the control group (n = 30). A total of 50 healthy single-birth full-term primiparous women and 30 women undergoing a second pregnancy were assigned to the maternal group, while 30 non-pregnant women matched by general data were assigned to the control group. Subject cognitive function was assessed using the Montreal Cognitive Assessment (MoCA) (Beijing version) and the Birmingham Cognitive Screen (BCoS) (Mandarin version); related influencing factors were analyzed. RESULTS: In the maternal group, the results showed a MoCA score of 26.52 ± 2.13 points and a cognitive impairment incidence of 26% in primiparous women, along with a MoCA score of 25.83 ± 2.49 points and a cognitive impairment incidence of 36.7% in women undergoing a second pregnancy. All scores were lower in the maternal group than in the control group, which had a MoCA score of 27.47 ± 1.28 points and cognitive impairment incidence of 6.7% (p < 0.05). The differences in MoCA score and cognitive impairment incidence between the primiparous sub-group and the second pregnancy sub-group were not statistically significant (p > 0.05). The visual space and executive function MoCA scale scores were lower in the maternal group than in the control group (p < 0.01). Furthermore, the scores were lower in the maternal group than in the control group in the following BCoS items: instant story recall, total apple deletion number, auditory attention, rule conversion, and gesture imitation (p < 0.05). CONCLUSION: Women in the postpartum period may develop cognitive dysfunction; however, the difference in cognitive impairment incidence between the primiparous sub-group and the second pregnancy sub-group in this study was not statistically significant. The educational level, labor analgesia, and total labor time (min) were found to be influencing factors in the postpartum cognitive function decline (p < 0.05).

Levels of vaccination coverage among HIV-exposed children in China: a retrospective study.

BACKGROUND: Vaccination is crucial for human immunodeficiency virus (HIV)-exposed children because of their increased risk of morbidity and mortality from various vaccine-preventable diseases. However, studies have shown that they are at high risk of incomplete vaccination. Although China has developed prevention of mother-to-child transmission (PMTCT) of HIV programs substantially over the past decades, few studies have investigated the immunization levels of Chinese HIV-exposed children. Therefore, we aimed to evaluate vaccination coverage and its associated factors among HIV-exposed children in China during 2016‒2018. METHODS: We conducted a retrospective cohort review of all cases of Chinese HIV-exposed children born between July 1, 2016 and June 30, 2018 recorded in the Chinese information system on PMTCT. The vaccination coverage indicators refer to the percentage of children who received recommended basic vaccines, including Bacillus Calmette-Guérin (BCG), hepatitis B (HepB), polio, measles-containing vaccine (MCV), and diphtheria-tetanus-pertussis-containing (DTP) vaccine. Univariate and multivariate logistic regression analyses expressed as crude odds ratios (cORs) and adjusted odds ratios (aORs), each with 95% confidence intervals (95% CI), were performed to compare the proportional differences of factors associated with vaccine coverage. RESULTS: Among the enrolled 10 033 children, the vaccination rate was 54.1% for BCG, 84.5% for complete HepB vaccination, 54.5% for complete polio vaccination, 51.3% for MCV, and 59.5% for complete DTP vaccination. Children with perinatally acquired HIV (PHIV) were 2.46‒3.82 times less likely to be vaccinated than HIV-exposed uninfected children. Multivariate logistic regression indicated that children of Han ethnicity (aOR = 1.33‒2.04), children with early infant diagnosis (EID) of HIV (aOR = 1.86‒3.17), and children whose mothers had better education (college or above, aOR = 1.63‒2.51) had higher odds of being vaccinated. Most of the deceased children (aOR = 4.28‒21.55) missed vaccination, and PHIV (aOR = 2.46‒3.82) significantly affected immunization. CONCLUSIONS: Chinese HIV-exposed children had low vaccination coverage, which is a serious health challenge that needs to be addressed thoroughly. Interventions should be developed with a focus on minority HIV-exposed children whose mothers do not have formal education. Particularly, more attention should be paid to EID to increase access to immunization.

Glipizide controlled-release tablets, with or without acarbose, improve glycaemic variability in newly diagnosed Type 2 diabetes.

1. The aim of the present study was to compare the effects of glipizide controlled-release (CR) tablets monotherapy with that of glipizide CR tablets plus acarbose on glycaemic variability in newly diagnosed Type 2 diabetes (T2DM) patients using a continuous glucose-monitoring system (CGMS). 2. Forty newly diagnosed T2DM patients whose glycated haemoglobin A1c (HbA1c) levels ranged from 7.0% to 9.8% were randomized to either monotherapy or combination therapy. Overall glycaemic control and blood glucose variability were evaluated by CGMS parameters. 3. After 8 weeks treatment, fasting and postprandial blood glucose, HbA1c, glycated albumin (GA), mean blood glucose (MBG), mean amplitude of glycaemic excursions (MAGE), postprandial incremental area under the curve (AUC(pp)) and homeostasis model assessment of insulin resistance decreased significantly in both groups (P < 0.01). There was also a significant decrease in the mean of daily differences (MODD) in the combination therapy group. Mean changes in MBG, MAGE, MODD and AUC(pp) were significantly greater in the combination therapy group than in the monotherapy group (all P < 0.01), whereas no significant differences were found in the mean changes of HbA1c and GA. Multivariate regression analysis showed that the decrement in AUC(pp) was significantly associated with decreases in MAGE. 4. In conclusion, glipizide CR tablets alone or in combination with acarbose can improve overall blood glucose levels and glycaemic variability. Combination therapy using glipizide CR tablets and acarbose was more effective in reducing intraday and day-to-day glycaemic variability than glipizide CR tablet monotherapy.

Prognostic models for acute liver failure.

BACKGROUND: Acute liver failure (ALF) remains a dramatic and unpredictable disease with high morbidity and mortality. Early and accurate prognostic assessment of patients with ALF is critically important for optimum clinical pathway. DATA SOURCES: Five English-language medical databases, MEDLINE, ScienceDirect, OVID, Springer Link and Wiley Interscience were searched for articles on "acute liver failure", "prognosis", and related topics. RESULTS: Multi-variable prognostic models including the King's College Hospital criteria and the model for end-stage liver disease score have been widely used in determination of the prognosis of ALF, but the results are far from satisfactory. Other prognostic indicators including serum Gc-globulin, arterial blood lactate, serum phosphate, arterial blood ammonia, and serum alpha-fetoprotein are promising but await further assessement. CONCLUSIONS: A reliable prognostic model to be developed in the future should not only have predictive value for poor outcome but also help to predict the survival of patients without a liver transplantation. Further studies are necessary to assess the prognostic accuracy of any new models.

Effects of plasma from patients with acute on chronic liver failure on function of cytochrome P450 in immortalized human hepatocytes.

BACKGROUND: The bioartificial liver is anticipated to be a promising alternative choice for patients with liver failure. Toxic substances which accumulate in the patients' plasma exert deleterious effects on hepatocytes in the bioreactor, and potentially reduce the efficacy of bioartificial liver devices. This study was designed to investigate the effects of plasma from patients with acute on chronic liver failure (AoCLF) on immortalized human hepatocytes in terms of cytochrome P450 gene expression, drug metabolism activity and detoxification capability. METHODS: Immortalized human hepatocytes (HepLi-2 cells) were cultured in medium containing fetal calf serum or human plasma from three patients with AoCLF. The cytochrome P450 (CYP3A5, CYP2E1, CYP3A4) expression, drug metabolism activity and detoxification capability of HepLi-2 cells were assessed by RT-PCR, lidocaine clearance and ammonia elimination assay. RESULTS: After incubation in medium containing AoCLF plasma for 24 hours, the cytochrome P450 mRNA expression of HepLi-2 cells was not significantly decreased compared with control culture. Ammonia elimination and lidocaine clearance assay showed that the ability of ammonia removal and drug metabolism remained stable. CONCLUSIONS: Immortalized human hepatocytes can be exposed to AoCLF plasma for at least 24 hours with no significant reduction in the function of cytochrome P450. HepLi-2 cells appear to be effective in metabolism and detoxification and can be potentially used in the development of bioartificial liver.

Calculating the Number of Pregnant Women Receiving Standardized Services for PMTCT of HIV in Liangshan Prefecture, Based on the Equivalent Method.

OBJECTIVE: To calculate the number of pregnant women who receive standardized prevention of mother-to-child transmission (PMTCT) services for HIV annually. METHODS: HIV-positive pregnant women in six counties of Liangshan Prefecture in 2017 were selected as study subjects. The entire process, from when the subjects first received the PMTCT of HIV services to the end, was divided into four stages, which were further divided into 25 phases. The equivalent coefficient was used to indicate the weight of workload in each phase. Seven experts were invited to score the equivalent coefficient; the number of pregnant women who received standardized services to prevent the transmission of HIV was calculated. RESULTS: A total of 663 HIV-positive pregnant women were registered in six Liangshan Prefecture counties in 2017. This figure was converted into 7,780 person-months devoted to HIV-positive pregnant women, with 260 person-months (3.34%) spent on the first antenatal care, 1,510 person-months (19.41%) during pregnancy, 378 person-months (4.86%) on delivery, and 5,632 person-months (72.39%) on post-partum period. The equivalent coefficient calculation showed that 314 HIV-positive pregnant women received standardized PMTCT services. CONCLUSION: The number of pregnant women receiving standardized services for the PMTCT of HIV can be calculated accurately using the equivalent method to identify the gap between the level of PMTCT of HIV intervention services needed and the actual workload.

Herpes simplex virus type I-infected disorders alter the balance between Treg and Th17 cells in recurrent herpes labialis patients.

Recurrent herpes labialis (RHL) is a common skin disease that is often caused by herpes simplex virus type I (HSV-1), but its immunology and pathogenesis remain unclear. The balance of Th17/Treg cells is crucial for maintaining immune homeostasis. This study aimed to investigate whether the balance of Th17/Treg cells and related cytokines may be a determinant occurrence in patients with RHL. This is a clinical experimental research based on clinical observation and analysis. We collected RHL patients from the outpatient clinic of the Department of Dermatology of Zhejiang Chinese Medical University (Hangzhou, China) in 2017, conducted questionnaire survey and signed informed consent. Peripheral blood was collected from 30 patients with RHL and 30 healthy volunteers. Flow cytometry was used to detect the percentages of Treg cells and Th17 cells. Protein microarrays coated with 20 cytokines related to T-cell subsets were performed. Enzyme-linked immunosorbent assay (ELISA) assay was conducted to further verify the expression levels of the cytokines that were screened by protein microarrays. Percentages of Th17/Treg cells in peripheral blood of RHL patients were significantly increased compared to those in healthy volunteers. The fold changes of GM-CSF, IL-4, TGF-ß, IL-12, IL-10, IL-17F, and TNF-α were significantly increased compared with healthy volunteers. In addition, the expression of IL-4, IL-10, and TGF-ß in the serum of RHL patients increased significantly. Our results indicated an imbalance of Th17/Treg cells in RHL, and this imbalance is probably an important factor in the occurrence, development, and recovery of RHL.