[Risk factors analysis of protein energy wasting in children with chronic kidney disease].

Objective: To analyze the clinical characteristics and risk factors of protein energy wasting (PEW) in children with chronic kidney disease (CKD). Methods: Clinical data of 231 children with chronic kidney disease hospitalized in Beijing Children's Hospital affiliated to Capital Medical University from January 2018 to January 2023 were retrospectively analyzed to explore the incidence of PEW. According to the diagnostic criteria of CKDPEW, they were divided into a CKDPEW group and a non PEW group. The comparison between the groups was performed by independent-sample t test and Chi-squared test, and the risk factors were analyzed by multivariate Logistic regression. Results: Among the 231 children, there were 138 males and 93 females, with a visiting age of 9.9 (7.9, 16.0) years; 6 cases were in stage 1, 14 cases in stage 2, 51 cases in stage 3, 36 cases in stage 4, and 124 cases in stage 5. A total of 30 children (13.0%) with CKD PEW were diagnosed at the age of 7. 1 (3.8, 13.2) years, including 1 case in stage 1, 1 case in stage 2, 5 cases in stage 3, 5 cases in stage 4, and 18 cases in stage 5. There were a total of 201 cases (87.0%) in the non PEW group, diagnosed at the age of 11.8 (8.5, 12.2) years, including 5 cases in stage 1, 13 cases in stage 2, 46 cases in stage 3, 31 cases in stage 4, and 106 cases in stage 5. The Chi-squared test and t test showed that the systolic blood pressure, diastolic blood pressure, birth weight and carbon dioxide binding capacity of the CKD PEW group were lower than those of the non PEW group ((109±22) vs. (120±20) mmHg (1 mmHg=0.133 kPa), (72±19) vs. (79±16) mmHg, (2.9±0.5) vs. (3.2±0.6) kg, (17±4) vs. (19±4) mmol/L,t=2.85, 2.14, 0.67, 2.63, all P<0.05). Multivariate logistic regression analysis showed that carbon dioxide binding capacity and birth weight were independent protective factors of CKDPEW in children (OR=0.81 and 0.36, 95%CI=0.73-0.90 and 0.17-0.77, respectively; both P<0.01); the risk of PEW in CKD children decreased by 0.187 times for every 1 mmol/L increment in carbon dioxide binding capacity, and 0.638 times for every 1 kg increment in birth weight. Conclusions: The incidence of protein energy expenditure in children with chronic kidney disease is lower than that in the previous researches. PEW can appear in CKD 1-2 stage, and attention should be paid to it in the early stage of CKD in clinical practice. Low birth weight CKD children are susceptible to PEW, and actively correcting metabolic acidosis can reduce the risk of CKDPEW.

Lateral retraction could achieve better early postoperative knee function than patellar eversion in total knee arthroplasty: a systematic review and meta-analysis.

OBJECTIVE: Comparisons between patellar eversion (PE) and lateral retraction (LR) in total knee arthroplasty (TKA) are still inconclusive. To determine the most suitable procedure, we aimed to evaluate the safety and efficacy of PE and LR in TKA in this meta-analysis. MATERIALS AND METHODS: This meta-analysis complied with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Web-based literature databases, including WANFANG, VIP, CNKI, the Cochrane Library, Embase, and PubMed, were utilized to conduct a comprehensive literature search for studies published until June 2022 that compared PE with LR in primary TKA. The quality of the selected randomized controlled trials (RCTs) was evaluated using guidelines of the Cochrane Reviews Handbook 5.0.2. RESULTS: A total of 10 RCTs, including 782 patients and 823 TKAs, were selected in this meta-analysis. Our results showed that using LR improved postoperative knee extensor function and range of motion (ROM). In addition, PE and LR resulted in similar clinical benefits in terms of Knee Society Function score, pain, length of hospital stay, Insall-Salvati ratio, the occurrence of patella baja, and complications related to the operation. CONCLUSIONS: Existing evidence suggested that using LR in TKA improved early postoperative knee function. Similar clinical and radiographic outcomes were obtained 1 year after the procedures were performed. Based on these findings, we recommended the use of LR in TKA. However, studies with large sample sizes are needed to validate these findings.

Aetiology of tinea capitis in China: a multicentre prospective study.

BACKGROUND: Tinea capitis is still common in developing countries, such as China. Its pathogen spectrum varies across regions and changes over time. OBJECTIVES: This study aimed to clarify the current epidemiological characteristics and pathogen spectrum of tinea capitis in China. METHODS: A multicentre, prospective descriptive study involving 29 tertiary hospitals in China was conducted. From August 2019 to July 2020, 611 patients with tinea capitis were enrolled. Data concerning demography, risk factors and fungal tests were collected. When necessary, the pathogens were further identified by morphology or molecular sequencing in the central laboratory. RESULTS: Among all enrolled patients, 74·1% of the cases were in patients aged 2-8 years. The children with tinea capitis were mainly boys (56·2%) and were more likely than adults to have a history of animal contact (57·4% vs. 35·3%, P = 0·012) and zoophilic dermatophyte infection (73·5% vs. 47%). The adults were mainly female (83%) and were more likely than children to have anthropophilic agent infection (53% vs. 23·9%). The most common pathogen was zoophilic Microsporum canis (354, 65·2%), followed by anthropophilic Trichophyton violaceum (74, 13·6%). In contrast to the eastern, western and northeastern regions, where zoophilic M. canis predominated, anthropophilic T. violaceum predominated in central China (69%, P < 0·001), where the patients had the most tinea at other sites (20%) and dermatophytosis contact (26%) but the least animal contact (39%). Microsporum ferrugineum was the most common anthropophilic agent in the western area, especially in Xinjiang province. CONCLUSIONS: Boys aged approximately 5 years were the most commonly affected group. Dermatologists are advised to pay more attention to the different transmission routes and pathogen spectra in different age groups from different regions.

[Efficacy and safety of percutaneous closure of ventricular septal rupture after acute myocardial infarction: a clinical study of 69 cases].

Objective: To investigate the efficacy and safety of percutaneous closure of ventricular septal rupture (VSR) after acute myocardial infarction (AMI) and the risk factors of all-cause mortality at 30 days after operation. Methods: This is a retrospective case series study. A total of 69 patients with post-AMI VSR, underwent percutaneous closure of VSR from October 2013 to May 2020 in Department of Cardiology of Henan Provincial People's Hospital and Department of Cardiology of Central China Fuwai Hospital, were included. Patients were divided into survival group (53 cases) and non-survival group (16 cases) according to the status at 30 days after operation. Clinical data were collected and analyzed during hospitalization. Telephone follow-up was performed 30 days after operation. The primary safety endpoint was occlusion failure and all-cause mortality at 30 days post operation. The secondary safety endpoint was the operation related or non-operation related complications. Efficacy endpoint included NYHA classification of cardiac function, index measured by right heart catheterization and echocardiography. Multivariate logistic regression was performed to analyze the risk factors of all-cause mortality at 30 days after operation. Results: A total of 69 patients, aged 67 (64, 71) years, including 42 women (60.9%), were enrolled in this study. All-cause death occurred in 16 patients (23.2%), including 13 in-hospital death and 3 death during follow-up. There were 4 cases of closure failure (5.8%). Among the 65 patients with successful closure, 12 (18.5%) experienced operation-related complications, among which 8 (12.3%) experienced valve injury. The mortality was significantly higher in patients with operation-related complications than that in patients without operation-related complications (41.7% (5/12) vs. 13.2% (7/53), P = 0.022). One case received percutaneous closure of VSR and PCI, this patient experienced new-onset AMI immediately post procedure and died thereafter (1.5%). One case (1.5%) developed multiple organ failure and 2 cases (3.1%) developed gastrointestinal bleeding post operation. All of the 65 patients with successful occlusion completed postoperative echocardiography, 56 patients completed cardiac function assessment at discharge, and 53 patients who survived up to 30 days post discharge completed clinical follow up by telephone. The NYHA cardiac function at discharge and 30 days after operation were significantly improved as compared to that before operation (P<0.001), the ratio of NYHA Ⅰ and Ⅱ patients was significantly higher post operation at these two time points as compared to baseline level (76.8% (43/56) vs. 23.1% (15/65), P<0.001, 77.4% (41/53) vs. 23.1% (15/65), P<0.001). The pulmonary circulation/systemic circulation blood flow ratio (Qp/Qs), pulmonary artery systolic pressure (PASP) and left ventricular end-diastolic diameter (LVDd) were decreased, aortic systolic pressure (ASP) and left ventricular ejection fraction (LVEF) were increased post operation (P<0.05). Multivariate logistic regression analysis showed that WBC>9.8×109/L (OR=20.94, 95%CI 1.21-362.93, P=0.037) and NT-ProBNP>6 000 ng/L (OR=869.11, 95%CI 2.93-258 058.34, P=0.020) were the independent risk factors of mortality at 30 days. Conclusions: Percutaneous closure in VSR after AMI is safe and effective. The increase of WBC and NT-ProBNP are the independent risk factors of all-cause mortality at 30 days after operation.

Long-term Outcomes of Endoscopic Resection for Gastric Subepithelial Tumors.

OBJECTIVE: The purpose of the current study was to analyze the safety and efficacy of endoscopic resection for gastric subepithelial tumors (SETs) using long-term patient outcome data. PATIENTS AND METHODS: A retrospective analysis of 73 consecutive patients with gastric SETs was performed from June 2014 to December 2016. The treatment methods included submucosal dissection, submucosal excavation or endoscopic full-thickness resection (EFTR). In addition to epidemiological data (sex and age), tumor size, surgical parameters, length of stay, complications, costs, and endoscopic, clinicopathologic, and follow-up data were analyzed to compare treatments. RESULTS: The complete resection rate was 97.3% (71/73). Three patients experienced complications (4.1%), including 2 with delayed perforation and 1 with perioperative infection. The median postoperative feeding time was 3 days, and the median postoperative hospital stay was 5 days. The median follow-up period was 19 months, with no patient death or tumor recurrence. Among the 38 patients with gastrointestinal stromal tumors, the complete resection rate was 97.4% (37/38). The complete resection and complication rates between the endoscopic submucosal excavation (ESE) group and the EFTR group were not statistically significant. There was no recurrence or metastasis detected among either group; however, the ESE group had earlier postoperative feeding, a shorter postoperative hospital stay, and less hospitalization expenses. CONCLUSIONS: Endoscopic resection for gastric SETs (<3 cm) is safe and feasible concerning medium-term and long-term effects. Compared with the EFTR group, the ESE group had earlier postoperative feeding, a shorter postoperative hospital stay, and less hospitalization expenses. Even so, gastric SETs with malignant potential are at risk of recurrence. Larger prospective multicenter studies are warranted.

The nuclear hormone receptor E75A regulates vitellogenin gene (Al-Vg) expression in the mirid bug Apolygus lucorum.

Apolygus lucorum is the predominant pest of Bacillus thuringiensis (Bt) cotton in China. 20-hydroxyecdysone (20E) plays a key role in the reproduction of this insect. To better understand the mechanism underlying 20E-regulated reproduction, the nuclear hormone receptor E75 isoform-A of Ap. lucorum (Al-E75A) was cloned and its expression analysed. A 2241-bp sequence of Al-E75A cDNA encoded an open reading frame of a polypeptide with a predicted molecular mass of 69.04 kDa. Al-E75A mRNA was detected in female adult stages of Ap. lucorum with peak expression in 7-day-old animals. Al-E75A was also expressed in several tissues, particularly in the fat body and ovary. A 3.2 kb Al-E75A mRNA was detected in all tissues by Northern blot. The fecundity and longevity were significantly decreased in female adults treated with Al-E75A small interfering RNA. The rates of egg incubation rates were considerably lower in the RNA interference-treated animals compared to the untreated controls. In order to investigate the molecular mechanism underlying the effects described above, vitellogenin (Al-Vg) was selected for further investigation. The expression pattern of Al-Vg was similar to that of Al-E75A and was up-regulated by 20E. After knockdown of Al-E75A, the expression profile of Al-Vg and the protein levels were down-regulated. These findings suggest that Al-E75A plays a crucial role in the regulation of Al-Vg expression in Ap. lucorum.

Oral immunization with a Lactobacillus casei-based anti-porcine epidemic diarrhoea virus (PEDV) vaccine expressing microfold cell-targeting peptide Co1 fused with the COE antigen of PEDV.

AIMS: The aims of this study were to develop an effective M cell-targeting oral vaccine, involving Lactobacillus casei to deliver the porcine epidemic diarrhoea virus (PEDV) core neutralizing epitope (COE) antigen conjugated with M cell-targeting peptide Co1 as an adjuvant, against PEDV infection. METHODS AND RESULTS: Genetically engineered L. casei 393 (L393) strains expressing PEDV COE antigen only (pPG-COE/L393) or fused-expressing COE and M cell-targeting peptide Co1 (pPG-COE-Co1/L393) were constructed, and the immunogenicity upon administration as an oral vaccine was evaluated. The results showed that higher anti-PEDV serum IgG and mucosal SIgA antibody responses were induced in mice orally immunized with strain pPG-COE-Co1/L393 as compared to the mice immunized with strain L393 expressing COE alone or carrying the empty plasmid. In addition, the use of the Co1 ligand elicited a splenocyte proliferative response more effectively in comparison with the COE antigen alone and supported a skewed T helper 2 type of immune response against PEDV. CONCLUSIONS: pPG-COE-Co1/L393 can effectively induce mucosal, humoural and Th2-type cellular immune responses against PEDV infection via oral administration. Furthermore, M cell-targeting peptide ligand Co1 is a good mucosal adjuvant. SIGNIFICANCE AND IMPACT OF THE STUDY: Lactobacillus casei delivering the COE antigen of PEDV conjugated with a M cell-targeting peptide Co1 as an immune adjuvant is a promising oral vaccine candidate for PEDV.

Virulence and serological studies of recombinant infectious hematopoietic necrosis virus (IHNV) in rainbow trout.

Infectious hematopoietic necrosis virus is a highly contagious disease of juvenile salmonid species. From the IHNV HLJ-09 isolated in China, two recombinant viruses were generated by reverse genetics using the RNA polymerase II transcription system. The recombinant viruses were confirmed by RT-PCR, indirect immunofluorescence assay and electron microscopy. They were referred to as rIHNV HLJ-09 and rIHNV-EGFP. rIHNV HLJ-09 and rIHNV-EGFP could stably replicate in EPC cell lines and had the same cellular tropism as wtIHNV HLJ-09. But the titer of rIHNV-EGFP was significantly lower than rIHNV HLJ-09 and wtIHNV HLJ-09. rIHNV-EGFP strain could express EGFP stably at least in 20 passages, and the fluorescence could be observed clearly. To assess the virulence and pathogenicity of the recombinant viruses in vivo, juvenile rainbow trout were challenged by intraperitoneal injection with 20µl of rIHNV HLJ-09, rIHNV-EGFP or wtIHNV HLJ-09 (1×10(6)pfuml(-1)). Fish challenged with rIHNV HLJ-09 and wtIHNV HLJ-09 exhibited clinical signs typical of IHN disease and both produced 90% cumulative percent mortality, whlie rIHNV-EGFP produced only 5%. Pathological sectioning results showed that the tissues (liver, kidney, heart muscle, back muscle) of the fish infected with rIHNV HLJ-09 exhibited pathological changes, with the exception of cerebral neurons and the cheek. However, no lesions of liver, kidney, heart, muscle, brain in rainbow trout of rIHNV-EGFP or the control group were observed. Indirect ELISA results showed that a high level of serum antibody was detected in the experimental fish challenged with rIHNV HLJ-09, just as the same as wtIHNV HLJ-09, while a lower titer was detecred in the fish infected with rIHNV-EGFP. This indicated that the recombinant viruses could induce humoral immune response in the experimental fish. The recombinant viruses had unique genetic tags and could be used for genetic engineering, laying new ground for further investigation of IHNV pathopoiesis molecular mechanism, host tropism and the development of novel vaccines against IHN.

Maternal obesity during pregnancy is negatively associated with maternal and neonatal iron status.

BACKGROUND/OBJECTIVES: Obesity among pregnant women may adversely affect both maternal iron status throughout pregnancy and placental transfer of iron. The objective of this study was to determine the association of maternal body mass index (BMI) with (1) maternal iron status and inflammation in mid and late pregnancy, (2) the change in maternal iron status throughout pregnancy and (3) neonatal iron status. SUBJECTS/METHODS: We examined longitudinal data from 1613 participants in a pregnancy iron supplementation trial in rural China. Women with uncomplicated singleton pregnancies were enrolled in the early second trimester of pregnancy and followed through parturition. Maternal blood samples obtained at enrollment and in the third trimester and cord blood samples were analyzed for a range of hematological and iron biomarkers. RESULTS: There was a negative association between maternal BMI and iron status at enrollment (transferrin receptor (sTfR): r=0.20, P<0.001; body iron (BI): r=-0.05; P=0.03). This association was markedly stronger among obese women. Maternal BMI was positively associated with maternal inflammation (C-reactive protein: r=0.33, P<0.001). In multiple linear regression models, maternal BMI was negatively associated with neonatal iron status (cord serum ferritin: -0.01, P=0.008; BI: -0.06, P=0.006) and associated with a lower decrease in iron status throughout pregnancy (sTfR: -4.6, P<0.001; BI: 1.1, P=0.004). CONCLUSIONS: Maternal obesity during pregnancy may adversely affect both maternal and neonatal iron status, potentially through inflammatory pathways.

Analysis of the genome sequence of infectious hematopoietic necrosis virus HLJ-09 in China.

Infectious hematopoietic necrosis virus (IHNV) is a highly contagious disease of juvenile salmonid fish. Six genome target fragments of the complete genome sequence of IHNV HLJ-09 were amplified by RT-PCR, and the 3'-terminal and 5'-terminal region of the genomic RNA were amplified using the RACE method. The complete genome sequence of HLJ-09 comprises 11,132 nucleotides (nt) (Accession number JX649101) and is different from that of other IHNV strains published in GenBank. Homology comparison and phylogenetic analysis of six ORF sequences were carried out using HLJ-09 and other IHNV strains published in GenBank. From phylogenetic tree analysis, the N gene, M gene, and P gene had the closest genetic relationship to IHNV-PRT from Korea. Phylogenetic analysis for the full length of the G gene showed that the HLJ-09 strain exhibited very close homology to the ChYa07, RtNag96, RtUi02, and RtGu01 strains from Korea and Japan, indicating that the HLJ-09 strain belonged to the genotype JRt. Ultimately, the Chinese IHNV HLJ-09 strain may have originated in Korea and Japan.

Development of a DNA microarray-based multiplex assay of avian influenza virus subtypes H5, H7, H9, N1, and N2.

Outbreaks of highly pathogenic avian influenza have caused considerable economic losses in the poultry industry and have also resulted in human deaths since 2004. Rapid subtyping of highly pathogenic avian influenza viruses(HPAIVs) in clinical specimens is a prerequisite of prompt control of disease and prevention of its spreading. In this study, we describe development of a DNA microarray-based detection and subtyping of HPAIVs in field samples. DNA copies of matrix (M) protein genes for the H5, H7, and H9 subtypes of hemagglutinin (HA) and the N1 and N2 subtypes of neuraminidase (NA) were prepared by RT-PCR and specific primers and then spotted onto aldehyde slides to form DNA microarrays. The HPAIV samples to be tested were subjected to total RNA isolation, RT-PCR with universal primers and Cy3 labeling, and the obtained double-stranded DNAs (targets) were finally hybridized with DNA microarrays (probes). A fluorescent spot on the microarray, detected by scanning indicated positive hybridization, i.e. the involved subtype. The assay was specific as various heterologous viruses or HPAIVs of other subtypes tested were negative. No cross-hybridization among different subtypes could be detected. The assay was more sensitive than RT-PCR and chicken embryo inoculation and could be also used for field samples. Summing up, the assay has proved useful for simultaneous detection and differentiation of main epidemic HPAIV subtypes.

In vivo analysis of kallikrein-related peptidase 6 (KLK6) function in oligodendrocyte development and the expression of myelin proteins.

Oligodendrocytes are important for not only nerve conduction but also central nervous system (CNS) development and neuronal survival in a variety of conditions. Kallikrein-related peptidase 6 (KLK6) is expressed in oligodendrocytes in the CNS and its expression is changed in several physiological and pathological conditions, especially following spinal cord injury (SCI) and experimental autoimmune encephalomyelitis. In this study, we investigated the functions of KLK6 in oligodendrocyte lineage cell development and the production of myelin proteins using KLK6-deficient (KLK6(-/-)) mice. KLK6(-/-) mice were born without apparent defects and lived as long as wild-type (WT) mice. There was no significant difference in the numbers of oligodendrocyte precursor cells and mature oligodendrocytes in the adult naive spinal cord between WT and KLK6(-/-) mice. However, there were fewer mature oligodendrocytes in the KLK6(-/-) spinal cord than in the WT spinal cord at postnatal day 7 (P7). Expression of myelin basic protein (MBP) and oligodendrocyte-specific protein/claudin-11, major myelin proteins, was also decreased in the KLK6(-/-) spinal cord compared with the WT spinal cord at P7-21. Moreover, after SCI, the amount of MBP in the damaged spinal cords of KLK6(-/-) mice was significantly less than that in the damaged spinal cords of WT mice. These results indicate that KLK6 plays a functional role in oligodendrocyte development and the expression of myelin proteins.

Cerebral magnetic resonance imaging of compressed air divers in diving accidents.

To investigate the characteristics of the cerebral magnetic resonance imaging (MRI) of compressed air divers in diving accidents, we conducted an observational case series study. MRI of brain were examined and analysed on seven cases compressed air divers complicated with cerebral arterial gas embolism CAGE. There were some characteristics of cerebral injury: (1) Multiple lesions; (2) larger size; (3) Susceptible to parietal and frontal lobe; (4) Both cortical grey matter and subcortical white matter can be affected; (5) Cerebellum is also the target of air embolism. The MRI of brain is an sensitive method for detecting cerebral lesions in compressed air divers in diving accidents. The MRI should be finished on divers in diving accidents within 5 days.

99mTc-TRODAT-1 SPECT study in early Parkinson's disease and essential tremor.

OBJECTIVE: The clinical differentiation between early Parkinson's disease (PD) and essential tremor (ET) could be difficult, therefore, the aim of this study was to investigate 99mTc-TRODAT-1 SPECT as an applicable tool in this field. METHODS: 99mTc-TRODAT-1 single photon emission computed tomography (SPECT) was performed in 10 healthy volunteers, 27 patients with idiopathic PD (Hoehn and Yahr 1-1.5) and 12 patients with ET. The ratio of striatal (99m)Tc-TRODAT-1 binding was calculated as the index of (striatum - occipital cortex)/occipital cortex. RESULTS: Compared with the striatal 99mTc-TRODAT-1 uptake in the ET group (0.49 +/- 0.07) or healthy controls (0.54 +/- 0.18), there was a significant decrease in the bilateral striatums of early PD, with a greater reduction in the contralateral striatum (0.27 +/- 0.08) than ipsilateral one (0.36 +/- 0.10, P < 0.01). Its sensitivity and specificity of differentiating early PD from ET was 96.4% and 91.7% respectively. CONCLUSION: 99mTc-TRODAT-1 SPECT can detect the dysfunction of nigrostriatal system in patients with early PD and provided a feasible tool to help differentiate early PD from ET.

Dual regulation of glycogen synthase kinase-3beta by the alpha1A-adrenergic receptor.

Catecholamines, acting through adrenergic receptors, play an important role in modulating the effects of insulin on glucose metabolism. Insulin activation of glycogen synthesis is mediated in part by the inhibitory phosphorylation of glycogen synthase kinase-3 (GSK-3). In this study, catecholamine regulation of GSK-3beta was investigated in Rat-1 fibroblasts stably expressing the alpha1A-adrenergic receptor. Treatment of these cells with either insulin or phenylephrine (PE), an alpha1-adrenergic receptor agonist, induced Ser-9 phosphorylation of GSK-3beta and inhibited GSK-3beta activity. Insulin-induced GSK-3beta phosphorylation is mediated by the phosphatidylinositol 3-kinase/Akt signaling pathway. PE treatment does not activate phosphatidylinositol 3-kinase or Akt (Ballou, L. M., Cross, M. E., Huang, S., McReynolds, E. M., Zhang, B. X., and Lin, R. Z. (2000) J. Biol. Chem. 275, 4803-4809), but instead inhibits insulin-induced Akt activation and GSK-3beta phosphorylation. Experiments using protein kinase C (PKC) inhibitors suggest that phorbol ester-sensitive novel PKC and Gö 6983-sensitive atypical PKC isoforms are involved in the PE-induced phosphorylation of GSK-3beta. Indeed, PE treatment of Rat-1 cells increased the activity of atypical PKCzeta, and expression of PKCzeta in COS-7 cells stimulated GSK-3beta Ser-9 phosphorylation. In addition, PE-induced GSK-3beta phosphorylation was reduced in Rat-1 cells treated with a cell-permeable PKCzeta pseudosubstrate peptide inhibitor. These results suggest that the alpha1A-adrenergic receptor regulates GSK-3beta through two signaling pathways. One pathway inhibits insulin-induced GSK-3beta phosphorylation by blocking insulin activation of Akt. The second pathway stimulates Ser-9 phosphorylation of GSK-3beta, probably via PKC.

Rapamycin-insensitive regulation of 4e-BP1 in regenerating rat liver.

In cultured cells, growth factor-induced phosphorylation of two translation modulators, p70 S6 kinase and eukaryotic initiation factor 4E-binding protein 1 (4E-BP1), is blocked by nanomolar concentrations of the immunosuppressant rapamycin. Rapamycin also attenuates liver regeneration after partial hepatectomy, but it is not known if this growth-suppressive effect is due to dephosphorylation of p70 S6 kinase and/or 4E-BP1. We found that partial hepatectomy induced a transient increase in liver p70 S6 kinase activity and 4E-BP1 phosphorylation as compared with sham-operated rats. The amount of p70 S6 kinase protein in regenerating liver did not increase, but active kinase from partially hepatectomized animals was highly phosphorylated. Phosphorylated 4E-BP1 from regenerating liver was unable to form an inhibitory complex with initiation factor 4E. Rapamycin blocked the activation of p70 S6 kinase in response to partial hepatectomy in a dose-dependent manner, but 4E-BP1 phosphorylation was not inhibited. By contrast, functional phosphorylation of 4E-BP1 induced by injection of cycloheximide or growth factors was partially reversed by the drug. The mammalian target of rapamycin (mTOR) has been proposed to directly phosphorylate 4E-BP1. Western blot analysis using phospho-specific antibodies showed that phosphorylation of Thr-36/45 and Ser-64 increased in response to partial hepatectomy in a rapamycin-resistant manner. Thus, rapamycin inhibits p70 S6 kinase activation and liver regeneration, but not functional phosphorylation of 4E-BP1, in response to partial hepatectomy. These results indicate that the effect of rapamycin on 4E-BP1 function in vivo can be significantly different from its effect in cultured cells.

[Triterpenoid saponins from Eclipta prostrata L].

AIM: To study the triterpenoid saponins in the Chinese traditional medicine Eclipta prostrata L.. METHODS: Column chromatography with silica gel and HPLC were employed for the isolation and purification. The molecular structures were determined on the basis of spectral analysis (IR, MS, 1HNMR, 13CNMR, HMQC and HMBC). RESULTS: Two new triterpenoid saponins, named eclalbasaponins XI (4) and XII (5), were obtained and their structures were elucidated as 3-O-[beta-D-glucopyranosyl(1-->2)-beta-D-glucopyranosyl]-16 alpha-ethoxy-olean-12-ene-28-oic acid-28-O-beta-D-glucopyranoside and 3-O-[(2-O-sulfuryl-beta-D-glucopyranosyl) (1-->2)-beta-D-glucopyranosyl]-echinocystic acid-28-O-beta-D-glucopyranoside, respectively, along with three known saponins, eclalbasaponins II (1), I (2) and III (3). CONCLUSION: Compounds 4 and 5 are new compounds, 1 and 5 induced morphological deformation of Pyricularia oryzae mycelia.

Studying the growth of cervical carcinoma nests and angiogenesis by immunostaining, quantitation and three-dimensional structural analysis.

OBJECTIVE: To analyze the relationship and mutual effect of the growth of cervical carcinoma nests and angiogenesis. STUDY DESIGN: Serial paraffin sections of cervical squamous carcinoma were stained in repeated order with hematoxylin-eosin (HE), immunostain for factor VIII-related antigen, type IV collagen and proliferating cell nuclear antigen (PCNA). Three-dimensional reconstructions were made, and the volumes of carcinoma nests, necrosis and microvessels were measured. RESULTS: Two types of cervical carcinoma nests were distinguished on the basis of their growth characteristics and vascularity (groups I and II). Group I nests: The carcinoma cells were proliferating actively, as determined by their morphology and PCNA staining. There were abundant microvessels. Some endothelial sprouts and cords penetrated the nests and then formed networks and new vessels. The volume ratio of microvessels, including sprouts and cords, to the nests was 0.6282:1. Group II nests: The center of these nests underwent necrosis. The peripheral cells were rather small, with no mitosis. The PCNA index was rather low; these nests grew very slowly. There were only a few surrounding microvessels with no endothelial sprouts or cords. The volume ratio of vessels to nest was 0.0657:1. CONCLUSION: Two types of cervical carcinoma nests show a close relationship and mutual effect of the growth of carcinoma nest and angiogenesis. Group I nests grow and develop well, with abundant microvessels. Thus, many tumor cells may be angiogenic and induce angiogenesis; growth of the nests seemed dependent on adequate neovascularization. Group II nests grow slowly, with a few microvessels; the center of the nests undergoes necrosis. The inadequate blood supply must be one of the important causes of necrosis. Considering that there must have been abundant neovascularization during their growth in the past, most of the microvessels must have been obliterated and then reabsorbed to make the remaining vessels so few.