Transcriptomic analysis reveals the inhibition of reproduction in rice brown planthopper, Nilaparvata lugens, after silencing the gene of MagR (IscA1).

MagR (IscA1) is a member of the iron-sulphur cluster assembly proteins, which plays vital roles in many physiological processes, such as energy metabolism, electron transfer, iron homeostasis, heme biosynthesis and physiologically magnetic response. Its deletion leads to the loss of mitochondrial DNA, inactivation of iron-sulphur proteins and abnormal embryonic development in organisms. However, the physiological roles of MagR in insects are unclear. This study characterized the effects and molecular regulatory mechanism of MagR gene silencing on the reproduction of brachypterous female adults of Nilaparvata lugens. After silencing the MagR gene using RNAi approach, the duration of reproductive period was shortened and the fecundity and hatchability reduced significantly. A total of 479 differentially expressed genes (DEGs) were identified for female adults after 2 days of dsRNA injection through RNA-sequencing technology, including 352 significantly upregulated DEGs and 127 significantly downregulated DEGs, among which 44 DEGs were considered the key genes involved in the effects of NlMagR silencing on the reproduction, revealing the regulatory mechanism of MagR at RNA transcription level and providing a new strategy for the control of N. lugens.

Lamivudine therapy during the second vs the third trimester for preventing transmission of chronic hepatitis B.

There are little data on the timing of initiating lamivudine therapy for preventing transmission of hepatitis B in highly viremic mothers. Between May 2008 and January 2015, we retrospectively enrolled mothers with HBV DNA >6 log10  copies/mL who received lamivudine during pregnancy, and we compared them to untreated mothers. The primary measurement was the vertical transmission rate. The secondary outcomes were the mothers' and infants' safety. Among 249 consecutive mothers enrolled, 66 and 94 received lamivudine during the second and third trimesters, respectively, and 89 were untreated. At delivery, maternal mean HBV DNA levels were significantly lower in mothers who received lamivudine (4.45 log10; vs 7.16 log10  copies/mL; P<.001). Lamivudine treatment was well tolerated. However, early treatment during the second trimester did not significantly increase the percentage of mothers achieving HBV DNA levels of <6 log10  copies/mL compared to those treated during the third trimester (98.5% vs 94.7%; P=.40). At the age of 28 weeks, the vertical transmission rates were significantly lower in the lamivudine-treated mothers vs in the untreated mothers (0% [0/160] vs 5.62% [5/89]; P<.001), but the rates were similar when comparing the two subgroups treated with lamivudine (0% [0/66] vs 0% [0/94], P>.05). The birth defect rates and mothers' and infants' adverse events were similar among the groups. Lamivudine treatment initiated in the second or third trimester for mothers with HBV DNA levels below 9 log10  copies/mL was equally safe and effective in preventing vertical transmission. Thus, lamivudine should be deferred until the third trimester to minimize foetal exposure and drug resistance.

Correlation between Cystatin C and retinopathy of type-two diabetes mellitus patients.

Diabetic retinopathy is one of most common diabetic microvascular complications. In recent years the incidence of the disease has increased, hence early diagnosis and treatment are of great importance. In order to find reliable biological indexes to diagnose and treat type-two diabetes mellitus promptly, this study focused on the correlation between Cystatin C (Cys C) and retinopathy of type-two diabetes mellitus patients. One hundred and eighty type-two diabetes mellitus patients and one hundred healthy controls (the control group) were chosen in this study. Of the patients ninety-eight patients had typetwo diabetes mellitus without retinopathy (non-diabetic retinopathy group) and eighty-two had typetwo diabetes mellitus with retinopathy (diabetic retinopathy group). Correlation of Cys C and typetwo diabetic retinopathy was analyzed by examining the waist-hip ratio, fasting blood glucose (FBG), total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), glycosylated hemoglobin (HbA1c), and Cys C of both groups. The results showed that FBG, TC, TG, LDL-C, HbA1c, Cys C in the type-two diabetes mellitus patients group were higher than those of the control group (P less than 0.05). Age, course of diabetes, FBG, HbA1c, and Cys C levels were statistically significant in both the DR group and NDR group (P less than 0.05). The result of logistic regression analysis indicates that there was a positive correlation between type-two diabetic retinopathy development and age, course of diabetes, and Cys C level (P less than 0.05). Thus, it can be seen that changes of Cys C levels can assist early diagnosis and treatment of diabetic retinopathy to some extent. The patients with high Cys C level, long course of diabetes, and old age are more likely to have diabetic retinopathy.

Predictive value of serum gamma-glutamyltransferase levels in patients with hepatocellular carcinoma.

Here we assessed the predictive value of gamma-glutamyltransferase (γ-GT) for the prognosis of patients with HCC and compared the γ-GT with other prognostic factors. We retrospectively analyzed outcomes for 858 patients first diagnosed with HCC. Cox univariate and multivariate analyses receiver operating characteristic (ROC) curve were used for the study of significance of prognostic factor. A Kaplan-Meier survival analysis was performed to assess the value of γ-GT as an HCC prognostic factor in different classifications of Barcelona Clinic Liver Cancer (BCLC) or Tumor Node Metastasis (TNM) and different levels of serum alpha fetoprotein (AFP). We showed patient survival rates were significantly associated with γ-GT as well as serum biological markers including absolute neutrophil count (ANC), absolute lymphocyte count (ALC), AFP. A γ-GT ≥ 75 U/L strongly indicated poor prognosis for HCC patients. The survival time of patients with γ-GT ≥ 75 U/L was significantly shorter in advanced BCLC and TNM stages and at any serum AFP level. All these results suggested that baseline γ-GT could effectively aid in determining the prognosis of patients with HCC, and the prognostic value of γ-GT ≥ 75 U/L was superior to that of Child-Pugh class, MELD stage, and serum AFP.

Clinical effect of intravitreous injection of triamcinolone acetonide in treating cystoid macular edema.

Cystoid macular edema (CME), a commonly seen sign for multiple fundus diseases, is able to induce visual deterioration. The incidence rate of CME is constantly increasing; however, the existing clinical treatments cannot achieve satisfactory curative effects. To explore the curative effect of intravitreous injection of triamcinolone acetonide (TA) in treating CME, this study carried out a clinical test on 39 patients (42 eyes) from The First Affiliated Hospital of Zhengzhou University who developed CME induced by central retinal vein occlusion (CRVO). All 42 eyes received intravitreous injection of 40 mg/ml TA (0.1 ml) and then were followed up for 11-23.5 months. Eyes were examined by slit-lamp microscope, fundus fluorescein angiography (FFA) and optical coherence tomography (OCT) and best corrected visual acuity (BCVA), and intraocular pressure (IOP) of those eyes were detected before and after treatment. Average vision of eyes was 0.1 before treatment, and the vision improved in one month (vision ≥ 0.2: 100%; vision ≥ 0.5: 42.9%) and three months (vision ≥ 0.2: 64.3%; vision ≥ 0.5: 21.4%) after treatment; but as time went on, the vision of some patients declined; at the last follow-up, patients with vision ≥ 0.2 accounted for 28.6% and those with vision ≥0.5 accounted for 7.1%; compared to before treatment, 71.4% patients had improved vision and the remaining 28.6% had declined vision. Some patients were observed with high IOP during treatment, and 7 eyes were found with secondary cataract in posterior capsule of lens at the last follow-up. Intravitreous injection of triamcinolone acetonide proved to have significant short-term curative effect on CEM which is non-sensitive to conventional therapies, but it is likely to induce high IPO and posterior capsular opacification.

Effect of Xin Mai Jia on atherosclerosis in rats.

We investigated the therapeutic effect of Xin Mai Jia (XMJ) on atherosclerosis (AS) in rats. Rat models of AS were established by peritoneally injecting vitamin D, feeding a high-fat diet, and inducing balloon injuries in rats. The stomachs of the rats were irrigated continuously for 10 weeks with XMJ. Blood lipid- and hemorheology-related indices of blood samples were detected. Pathological changes in the right common carotid arterial tissues were also determined. The protein expression levels of endothelial nitric oxide synthase, angio-tensin-1, and endothelin-1 were determined by western blotting. XMJ reduced cholesterol, trigylecride, and low-density lipoprotein levels as well as blood viscosity, sedimentation, and hematocrit. Furthermore, XMJ alleviated vascular endothelial injury and reduced/eliminated atherosclerotic plaques. In contrast, XMJ significantly increased the endothelium-dependent relaxing response of the AS rat models. The western blotting results showed that XMJ upregulated endothelial nitric oxide synthase but downregulated angiotensin-1 and endothelin-1. XMJ prevented the development of AS by regulating blood lipid levels, hemorheology, and vascular function.

Neutrophil-lymphocyte ratio: a novel predictor for short-term prognosis in acute-on-chronic hepatitis B liver failure.

Acute-on-chronic hepatitis B liver failure (ACHBLF) has a poor prognosis in patients with hepatitis B virus infection. The role of the neutrophil-lymphocyte ratio (NLR), which reflects the inflammatory status of the patient before treatment, has never been studied in this setting. To investigate the predictive value of NLR in patients with ACHBLF, a retrospective cohort with 216 patients and a prospective validation cohort with 73 patients were recruited. Multivariate analyses showed that total bilirubin (TBIL), NLR, age and model for end-stage liver disease (MELD) score had prognostic significance for survival. Both NLR (0.781) and MELD score (0.744) had higher ROC curves, which differed significantly from those for age (0.615) and TBIL (0.691), but not from each other (P = 0.94). NLR ≤ 2.36 predicted lower mortality (with 91.6% sensitivity and 86.0% negative predictive value), and NLR >6.12 was a warning sign for higher mortality risk (with 90.1% specificity and 80.3% positive predictive value). These results demonstrated that pretreatment NLR was associated with the prognosis of patients with ACHBLF, and elevated NLR predicted poor outcome within 8 weeks. We suggest that NLR cut-offs of ≤ 2.36 and >6.12 are powerful markers for predicting mortality in ACHBLF.

Protective effect of ultrafiltered XinMaiJia extract against H2O2-induced injury in human umbilical vein endothelial cells through NHE1 downregulation.

We examined the protective effects of ultrafiltered XinMaiJia (XMJ) extract in a hydrogen peroxide (H2O2)-induced injury model in human umbilical vein endothelial cells (HUVECs) and determined the corresponding changes in the Na(+)-H(+) exchanger (NHE1) protein content and NHE1 gene expression. H2O2-induced HUVECs were treated with different concentrations of XMJ extract and the corresponding changes in morphology, activity, membrane permeability, biochemical indicators, cytokine concentration, NHE1 protein content, and NHE1 gene expression were determined. H2O2 significantly promoted HUVEC injury, whereas ultrafiltered XMJ extract significantly improved the morphological changes in injured HUVECs, increased their activity, and decreased NHE1 gene and protein expression, as well as limited the decrease in membrane permeability and expression of intercellular adhesion molecule-1, vascular cell adhesion molecule-1, interleukin (IL)-1, IL-6, and nuclear factor-kB. Ultrafiltered XMJ extract inhibited H2O2-induced HUVEC injury by inhibiting NHE1 activity.

Wenshen Xiaozheng Tang suppresses the growth of endometriosis with an antiangiogenic effect.

OBJECTIVE: To evaluate the effect of Wenshen Xiaozhng Tang (WXT) on ectopic endometrial growth and on angiogenesis in endometrial implants in a rat model. METHODS: Sprague-Dawley rats with endometriotic implants were randomly treated with low-dose WXT, high-dose WXT, or vehicle (negative control) for 28 days. Cell proliferation and vascular density in the lesions were assessed by immunohistochemistry. The levels of VEGF in peritoneal fluid were determined by ELISA. The mRNA expression of HIF-1α and Flk-1 in the endometriotic lesions was evaluated by real-time PCR. RESULTS: WXT treatment significantly decreased the lesion size and inhibited cell proliferation in the endometriotic lesions. Lowered vascular density and reduced mRNA expression of HIF-1α in the endometriotic lesions, associated with decreased concentration of VEGF in peritoneal fluid, were also observed in WXT-treated rats. CONCLUSIONS: These results suggest that WXT could be effective to suppress the growth of endometriosis, partially through its antiangiogenic activity.

Application of simulated puncture in percutaneous nephrolithotomy.

OBJECTIVE: This study aimed to explore the clinical value of simulated puncture in percutaneous nephrolithotomy in the treatment of complex kidney stones. PATIENTS AND METHODS: A total of 120 patients with complex kidney stones who were treated with percutaneous nephrolithotomy in our hospital between March 2017 and March 2020 were enrolled in this study and randomly divided into two groups: the research group and the control group (n = 60 in each). Each subject underwent a dual-source computed tomography (CT) scan of the pelvis and both kidneys before the operation. The research team imported the CT data into Mimics19 software to create a three-dimensional (3D) reconstruction of the skin, bones, kidneys, collecting system, and stones. Based on the 3D reconstruction model, the target renal calyx to be punctured was determined, the best puncture channel was designed, and puncture was simulated. Data regarding the simulated puncture were imported into 3-Matics11 software; the angle and depth of the puncture were measured, and then these data were used to guide percutaneous nephrolithotomy. 3D reconstruction and simulated puncture were not undertaken for the patients in the control group before the operation. The effects of treatment in the two groups were compared. RESULTS: First-stage percutaneous nephrolithotomy was successfully completed in both groups of patients. The outcome was better in the research group than in the control group in terms of operation time, number of punctures required for successful establishment of a percutaneous renal channel, number of percutaneous kidney puncture channels, and intraoperative blood loss, and the differences were statistically significant (p < 0.05 for all). The stone clearance rate was higher in the research group than in the control group, but the difference was not statistically significant (p = 0.471). The incidence of penetrating kidney injury was lower in the research group than in the control group, but the difference was not statistically significant (p = 0.154). CONCLUSIONS: For patients due to undergo percutaneous nephrolithotomy for the treatment of complex kidney stones, preoperative simulated puncture helps to improve the puncture accuracy and to reduce the number of punctures required for successful establishment of a percutaneous renal channel, the number of puncture channels, the operation time, and the blood loss, and therefore it is worth promoting.

Type 2 diabetes and impaired glucose tolerance in North-China-based rural community adults.

OBJECTIVES: To estimate the prevalence of type 2 diabetes and impaired glucose tolerance (IGT), and to identify potential risk factors among adults in rural areas in North China. STUDY DESIGN: Multistage random cluster sampling in a cross-sectional study. METHODS: Data were collected in 2005 from 1058 adults aged >20 years. Body mass index (BMI), waist and hip circumferences, 2-hour postprandial glucose (2-hPG) and other data were collected. RESULTS: The prevalence of diabetes and IGT was higher for both men and women in the study group compared with the national averages (diabetes: 7.17% and 7.01%, respectively; IGT: 7.55% and 7.95%, respectively). Men aged ≥60 years and women aged 50-59 years had the highest prevalence of diabetes compared with other age groups (9.62% and 9.21% respectively). Both men and women aged ≥60 years had the highest prevalence of IGT. A sudden increase in 2-hPG level was seen in women aged ≥40 years. Those with BMI ≥28 kg/m(2) were at two-fold higher risk than normal. The risk for glucose tolerance abnormalities was almost 1.55-fold higher in subjects with waist/height ratio (WHtR) ≥0.50. The odds ratio for diabetes was 0.32 (95% confidence interval 0.10-0.98) in subjects with an annual family income ≥30,000 yuan compared with those with lower incomes. CONCLUSIONS: These results indicate that rural populations in North China have a higher prevalence of type 2 diabetes and IGT compared with national averages. Women aged ≥40 years warrant more attention to avoid glucose tolerance abnormalities. BMI and WHtR are predictive of the prevalence of glucose tolerance abnormalities. High annual family income appears to be a protective factor for type 2 diabetes.

Long noncoding RNA ZFPM2-AS1 promotes the tumorigenesis of renal cell cancer via targeting miR-137.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "Long noncoding RNA ZFPM2-AS1 promotes the tumorigenesis of renal cell cancer via targeting miR-137, by J.-G. Liu, H.-B. Wang, G. Wan, M.-Z. Yang, X.-J. Jiang, J.-Y. Yang, published in Eur Rev Med Pharmacol Sci 2019; 23 (13): 5675-5681-DOI: 10.26355/eurrev_201907_18304-PMID: 31298319" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/18304.

Long noncoding RNA ZFPM2-AS1 promotes the tumorigenesis of renal cell cancer via targeting miR-137.

OBJECTIVE: Recently, long noncoding RNAs (lncRNAs) have attracted more attention for their roles in tumor progression. The aim of this study was to investigate the role of lncRNA ZFPM2 antisense RNA 1 (ZFPM2-AS1) in the progression of renal cell cancer (RCC), and to explore the possible underlying mechanism. PATIENTS AND METHODS: Expression levels of ZFPM2-AS1 in both RCC cells and 50 paired tissue samples were detected by Real Time-quantitative Polymerase Chain Reaction (RT-qPCR). Moreover, the relationship between lncRNA ZFPM2-AS1 expression level and clinic-pathological characteristics as well as patients' disease-free survival rate was explored, respectively. Furthermore, cell proliferation assay, wound healing assay and transwell assay were performed to investigate the role of lncRNA ZFPM2-AS1 in vitro. In addition, Western blot assay, Luciferase reporter gene assay and RNA immunoprecipitation assay were used to explore the possible underlying mechanism. RESULTS: The expression level of ZFPM2-AS1 in tumor tissues was significantly higher than that of corresponding normal tissues. ZFPM2-AS1 expression was associated with lymph node metastasis, tumor stage and survival time of RCC patients. Moreover, the overexpression of ZFPM2-AS1 significantly promoted the growth, invasion and migration of tumor cells, whereas remarkably inhibited cell apoptosis in vitro. Further experiments revealed that miR-137 was a direct target of ZFPM2-AS1. In addition, miR-137 expression in tumor tissues was negatively correlated with ZFPM2-AS1 expression. CONCLUSIONS: Our findings indicated that ZFPM2-AS1 could promote metastasis and proliferation, whereas inhibiting the apoptosis of RCC via targeting miR-137. This study might provide a new vision for interpreting the mechanism of RCC development.

[In vitro and in vivo effects of 5-aminolevulinic acid-mediated photodynamic therapy against oral squamous cell carcinoma].

Objective: To investigate the in vitro and in vivo effects of 5-aminolevulinic acid (5-ALA)-mediated photodynamic therapy against oral squamous cell carcinoma (OSCC) and preliminarily explore the possible mechanisms. Methods: SCC25 cells were divided into the control group (5-ALA of 0 mg/L) and the experimental group (5-ALA of 10, 25, 50, 100 and 150 mg/L). The production of protoporphyrin Ⅸ (PpⅨ) induced by 5-ALA in SCC25 cells was detected using the flow cytometry. SCC25 cells were divided into the control group (5-ALA of 0 mg/L), lazer alone group, 5-ALA alone group (5-ALA of 100 mg/L) and the 5-ALA combined with laser irradiation group (5-ALA of 5, 10, 25, 50 and 100 mg/L), the cytotoxicity of 5-ALA combined with laser irradiation (wave length 635 nm, power density 87 mW/cm(2) and laser dose 10.4 J/cm(2)) was evaluated in SCC25 cells using the methyl thiazolyltetrazolium assay (incubation times of 4, 8 and 12 h in each group) and the induction effect of combination treatment on the cell apoptosis was assessed by the flow cytometry (incubation time of 12 h in each group). The intracellular production of reactive oxygen species (ROS) triggered by 5-ALA combined with laser irradiation was determined using a fluorescence probe method (incubation time of 12 h in each group). A mouse OSCC xenograft model bearing SCC25 tumor was built, and the mice were divided into control group (saline), 5-ALA group (5-ALA of 50 mg/kg) and 5-ALA combined with laser irradiation group (5-ALA of 10, 25 and 50 mg/kg). Antitumor effect of 5-ALA combined with laser irradiation (wave length 635 nm, power density 158 mW/cm(2) and laser dose 94.8 J/cm(2)) was further measured. Results: 5-ALA induced the production of PpⅨ in SCC25 cells in a drug concentration (0-150 mg/L)-and incubation time (0-24 h)-dependent manner. When the 5-ALA concentration was 100 mg/L, the intracellular PpⅨ production was in a relatively stable state. Cell viability and apoptosis rate of 5, 10, 25, 50, 100 mg/L 5-ALA combined with laser irradiation are, respectively, (82.3±5.2)%, (3.13±0.38)%; (74.6±9.3)%, (5.38±0.55)%; (38.3±9.7)%, (17.97±2.72)%; (9.2±3.8)%, (24.47±3.37)%; (7.2±0.8)%, (43.01±5.96)%, which indicated that 5-ALA combined with laser irradiation notably inhibited the growth of SCC25 cells and also induced significant cell apoptosis compared with the control group [(96.3±6.0)%, (0.35±0.13)%, P<0.05]. After combination treatment (5-ALA of 5, 10, 25, 50 and 100 mg/L combined with laser irradiation, the mean fluorescence intensity of dichlorofluorescein is (1.46±0.12)×10(4), (2.16±0.30)×10(4), (3.57±0.34)×10(4), (81.70±13.05)×10(4), (113.00±7.35)×10(4), respectively, a large amount of ROS was produced in SCC25 cells compared with the control group [(0.96±0.15) ×10(4), P<0.05], which was in positive correlation with the intracellular PpⅨ content. 5-ALA (concentration of 10, 25 and 50 mg/kg) combined with laser irradiation greatly suppressed the tumor growth in SCC25 tumor-bearing mice compared to the control group (P<0.05). Conclusions: 5-ALA-mediated photodynamic therapy can trigger the generation of intracellular ROS that has significant cytotoxicity and apoptosis induction effect, and thus inhibit the tumor growth both in vitro and in vivo.

[Targeted-knockdown of Yes-associated protein inhibits the Warburg effect and the invasion of laryngeal cancer cells].

Objective: To investigate the migration and invasion behaviors of Hep-2 after the targeted knockdown of yes-associated protein (YAP). Methods: Hep-2 cells were knock-downed for YAP by shRNA as YAP-shRNA group, Hep-2 treated with non-specific shRNA as YAP-NC group, and Hep-2 with no treatment as control. Glucose uptake and lactate production in the cells were examined to assess Warburg effect. The migration and invasion behaviors of cells in three groups were observed. The expressions of vimentin and E-cadherin were detected by RT-PCR and Western Blot. The statistical software GraphPad Prism 7.0 was used to analyze significance of data. Two tailed Student' s t-tests was used to determine significance when only two groups were compared. P values of less than 0.05 was considered statistically significant. Results: Downregulation of YAP led to a obvious decrease in glucose uptake [(18.51±1.72)%] and lactate production [103.40±8.32] in Hep-2 cells compared with control [(41.20±1.11)% and 743.69±19.49, t=19.20 and 52.33, respectively, both P<0.01] and YAP-NC group [(39.60±0.78)% and 705.22±17.20, t=19.34 and 54.56, respectively, both P<0.01]. Compared with the control group (78.32±4.04) and YAP-NC group (77.28±3.11), the scratch healing ability of Hep-2 cells was significantly decreased in YAP-shRNA group (44.71±4.68). The P value was less than 0.01 (t=9.42 and 10.04). The number of cells with YAP-shRNA (33.30±4.19) passing through compartments was remarkable fewer than the control group (133.71±6.72) and YAP-NC group (126.32±4.21). The P value was less than 0.01 (t=21.96 and 27.13). The expression of E-cadherin protein in cells of YAP-shRNA group (6.16±0.11) was up-regulated compared with control (0.97±0.10, t=35.70, P<0.01) and YAP-NC group (1.13±0.09, t=36.28, P<0.01), while the expression of vimentin protein in cells of YAP-shRNA group (1.08±0.09) was down-regulated compared with control (5.67±0.12, t=29.91, P<0.01) and YAP-NC group (5.51±0.12, t=29.04, P<0.01). Conclusions: The down-regulation of YAP in Hep-2 inhibits the migration and invasion of cells via suppressing Warburg and EMT program.

Pharmacokinetic parameters of chlorzoxazone and its main metabolite, 6-hydroxychlorzoxazone, after intravenous and oral administration of chlorzoxazone to liver cirrhotic rats with diabetes mellitus.

Protein expression of the hepatic CYP2E1 has been reported to be increased in diabetic rats. This enzyme is the primary metabolizer of chlorzoxazone (CZX) to 6-hydroxychlorzoxazone (OH-CZX). Although patients with liver cirrhosis have a higher prevalence of diabetes mellitus, there have been no reported studies on the protein expression of CYP2E1 in rats induced to have liver cirrhosis and diabetes mellitus by injection of N-dimethylnitrosamine followed by streptozotocin [liver cirrhosis with diabetes mellitus (LCD) rats]. Thus, in the present study, the pharmacokinetics of CZX and OH-CZX were evaluated in LCD rats. Compared with control rats, LCD rats had significantly decreased (by 62%) total liver protein and significantly increased (by 124%) protein expression of CYP2E1, but the intrinsic clearance (Cl(int); formation of OH-CZX per milligram protein) was comparable in both groups of rats. As a result, the relative Cl(int) was also comparable for the two groups. Thus, OH-CZX formation in LCD and control rats was expected to be similar. As expected, after i.v. (20 mg/kg) and p.o. (50 mg/kg) administration of CZX, the area under the curve (AUC) of OH-CZX was comparable in control and LCD rats (i.v., 571 +/- 85.8 and 578 +/- 413 microg x min/ml, respectively; p.o., 1540 +/- 338 and 2170 +/- 1070 microg x min/ml, respectively). In LCD rats, the AUC(OH-CZX)/AUC(CZX) ratio was similar to the value in control rats after i.v. and p.o. administration. These results indicate that OH-CZX can be used as a chemical probe to assess the activity of CYP2E1 in LCD rats.

Human Delta1-pyrroline-5-carboxylate synthase: function and regulation.

Mammalian Delta(1)-pyrroline-5-carboxylate synthase (P5CS) is a bifunctional ATP- and NAD(P)H-dependent mitochondrial enzyme that catalyzes the coupled phosphorylation and reduction-conversion of L: -glutamate to P5C, a pivotal step in the biosynthesis of L: -proline, L: -ornithine and L: -arginine. Previously, we reported cloning and characterization of two P5CS transcript variants generated by exon sliding that encode two protein isoforms differing only by a two amino acid-insert at the N-terminus of the gamma-glutamyl kinase active site. The short form (P5CS.short) is highly expressed in the gut and is inhibited by ornithine. In contrast, the long form (P5CS.long) is expressed ubiquitously and is insensitive to ornithine. Interestingly, we found that all the established human cell lines we have studied expressed P5CS.long but not P5CS.short. In addition, expression of P5CS.long can be modulated by hormones: downregulation by hydrocortisone and dexamethasone and upregulation by estradiol, for example. Using a quantitative proteomic approach, we showed that P5CS.long is upregulated by p53 in p53-induced apoptosis in DLD-1 colorectal cancer cells. Functional genomic analysis confirmed that there are two p53-binding consensus sequences in the promoter region and in the intron 1 of the human P5CS gene. Interestingly, overexpression of P5CS by adenoviruses harboring P5CS.long or P5CS.short in various cell types has no effect on cell growth or survival. It would be of importance to further investigate the role of P5CS as a p53 downstream effector and how P5CS.short expression is regulated by hormones and factors of alternative splicing in cells isolated from model animals.