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To determine the physicians'compliance of hour-1 bundle for sepsis. A management system of hour-1 bundle for sepsis was established. The clinical data of 286 sepsis patients were collected, who were classified into 3 months before the bundle (control group), 9 months during process (observation group) and 3 months after bundle (study group). The compliance of hour-1 bundle implementation was compared in three groups. The results showed that with the application and implementation of the management system, the compliance of hour-1 bundle for sepsis in the control group, observation group and study group was 58.3%(28/48), 69.1%(105/152) and 88.4%(76/86) respectively (χ探讨医护人员执行脓毒症1 h集束化治疗管理体系的依从性。构建脓毒症1 h集束化治疗执行管理体系，收集构建前3个月、构建过程（9个月）及构建完成后3个月的286例脓毒症患者，分析执行管理体系构建前（对照组）、构建过程（观察组）及构建后（实施组）医护人员对1 h集束化治疗的依从性。结果显示，随着执行管理体系的应用及实施，医护人员对脓毒症1 h集束化治疗的依从性对照组为58.3%，观察组为69.1%，实施组为88.4%，差异有统计学意义（χ²=7.053，

为探讨上颌骨囊肿开放术后避免囊肿开窗口闭锁的改进方法，本文回顾性分析了2015—2020年于山东大学齐鲁医院行手术治疗的22例上颌骨囊肿患者的临床资料。其中含牙囊肿10例，上颌骨囊肿12例。采用改良泪前隐窝入路，延长下鼻甲前缘纵切口，切除上颌窦内侧壁及鼻腔底壁，将囊肿开放于鼻腔，拔除囊肿内牙齿，使用黏膜瓣消除裸露骨面。所有患者术后囊肿均保持开放。.

The application of day surgery on thoracic surgery is just started, and the innovation of surgical robots and their application on thoracic surgery bring new opportunities to the development of thoracic day surgery. However, the clinical practice of robot-assisted thoracic day surgery (RTDS) in China still has challenges and disagreements. Based on the experience of domestic experts in the field of RTDS clinical practice, this review discussed several key points of RTDS, including the future direction of RTDS, adjusting the indications according to their own conditions for the institutions carrying out RTDS, the robot-assisted advantage of RTDS being brought into play during the operation, and the perfect post-discharge follow-up mechanism being an important guarantee for the safe development of RTDS, to promote the application progress of RTDS in China.日间手术在胸外科领域的应用目前仍处于起步阶段，而近年来机器人辅助胸腔镜手术在胸外科的应用为胸外科日间手术的推广普及带来了新的契机。但目前国内机器人胸外科日间手术（RTDS）的临床应用仍存在诸多挑战与分歧。本文立足国内相关领域专家在RTDS临床实践中的心得体会，讨论了RTDS目前的焦点问题，从RTDS未来方向、开展RTDS的机构可根据自身条件调整适应证、减少创伤、严格出院标准、完善出院后随访机制5个方面予以探讨，以期促进RTDS在国内应用的普及与进步。.

Prostate cancer (PCa) is one of the most diagnosed cancers in men worldwide. Several studies have identified that circular RNAs (circRNAs) have a crucial impact on the biological processes in PCa. Therefore, it is necessary to study the molecular mechanism of circRNAs in tumor progression and metastasis.RNA interference was used to decrease circHIPK3 and MTDH expression. Overexpression vector was used to increase circHIPK3 and MTDH expression. Luciferase reporter assay were used to detect the relationship between circHIPK3 and miR-448 or between miR-448 and MTDH. MTT assay, colony formation assay and transwell assay were used to measure proliferation and migration of PCa cells.Circular RNA circHIPK3 was significantly increased in PCa tissues and cell lines. And overexpression of circHIPK3 promoted the migration, proliferation, and invasion of PC-3 and 22Rv1 cells, while knockdown of circHIPK3 markedly repressed the above-mentioned series of biological processes. Furthermore, circHIPK3 promoted metadherin (MTDH) expression by sponging miR-448. In vivo experiments, it was also found that overexpression of circHIPK3 significantly promoted tumor growth.Our research shows that circHIPK3 plays a carcinogenic effect in PCa by regulating the miR-448/MTDH axis, indicating that circHIPK3 may be a potential therapeutic target for PCa.

鼻窦胆脂瘤在临床上较为罕见，本文介绍了1例上颌窦胆脂瘤病例，总结其病例特点，并介绍了经泪前隐窝入路切除的治疗效果及经验。发现经泪前隐窝入路切除上颌窦胆脂瘤优势明显，可最大程度地暴露上颌窦窦腔，便于手术操作，彻底清除病变，治疗效果好，创伤小，具有一定的临床应用价值。.

To investigate the protective effect of ticagrelor on the myocardium of patients with ST-elevated acute coronary syndrome accompanied by diabetes mellitus.210 patients with diabetes mellitus receiving emergency percutaneous coronary intervention (PCI) due to ST-elevated acute coronary syndrome from December 2014 to June 2018 in the Hospital were selected and randomly divided into ticagrelor group and clopidogrel group. The myocardial microcirculation perfusion was evaluated via ST-segment elevation resolution (STR) in electrocardiogram (ECG) and myocardial blush grade (MBG). Myocardial necrosis markers, including creatine kinase (CK), CK-MB, and cardiac troponin I (cTnI), were evaluated. Moreover, the cardiac function was assessed using brain natriuretic peptide (BNP) level and left ventricular ejection fraction (LVEF). Finally, patients were followed up for one month on average, and the adverse cardiovascular and bleeding events were recorded.The results showed that CK, CK-MB, cTnI, and BNP levels in ticagrelor group were lower than those in clopidogrel group, and the differences were statistically significant (p0.05). Thrombolysis in myocardial infarction (TIMI) flow grading after the operation had no statistically significant difference between the two groups, and the usage rate of tirofiban in ticagrelor group was lower than that in clopidogrel group (p0.05). Besides, the myocardial microcirculation perfusion level after the operation in ticagrelor group was significantly higher than that in clopidogrel group. The proportions of STR ≥50% in ECG and MBG2 in ticagrelor group were significantly higher than those in clopidogrel group (p0.01). The incidence rate of mild bleeding in ticagrelor group was higher than that in clopidogrel group (p0.05).The application of ticagrelor in the treatment of ST-elevated acute coronary syndrome accompanied by diabetes mellitus can increase the level of myocardial microcirculation perfusion and improve the left heart function.

The presented study aimed to investigate the antitumor efficacy of combination of oxaliplatin with rapamycin, an mTOR inhibitor, in hepatocellular carcinoma (HCC). The activation status of mTOR pathway was first examined in HCC cell lines HepG2, BEL7402, and HuH7 using Western blotting. Effects of rapamycin, oxaliplatin, and their combination on the proliferation of HCC cells were determined in vitro using MTT assay and in vivo using a nude mice model bearing HepG2 xenografts. Drug-induced cell apoptosis was examined by flow cytometry. Expression of apoptosis-related protein was determined by Western blotting. We observed that mTOR pathway was activated in all three cell lines used in the current study. MTT assay demonstrated that oxaliplatin in combination with rapamycin synergistically inhibited the proliferation of HCC cells. The combination regimen reduced terminal tumor burden more efficiently than the corresponding monotherapy. The percentages of apoptotic cells and the expression levels of apoptosis-related proteins including cleaved caspase-9, -3, and PARP were significantly higher in combination-treatment groups than those in mono-drug-treatment groups. The ratios of Bax/Bcl-2 in cells exposed to both oxaliplatin and rapamycin were significantly increased compared to those in cells subjected to oxaliplatin or rapamycin alone treatment. Results obtained in the presented study suggested that combination of oxaliplatin and rapamycin was superior to mono-drug and may have a potential value in treatment of HCC.

This work studies the expression differences of YKL-40 and TLR4 in nasal sinus mucosa of chronic sinusitis patients with and without nasal polyps and its clinical significances. Fifty chronic sinusitis patients with nasal polyps and 50 chronic sinusitis patients without, accepted by our hospital during February 2016-February 2017, were included and taken as group A and group B, respectively. In addition, another 50 patients with nasal deviation were taken as group C (control group). The ostiomeatal complex mucosa of group A and B and the inferior turbinate mucosa of group C were taken and the fluorescence quantitative PCR method was applied to detect the expression of YKL-40, TLR4 and NF- κB of the mucosa and explore and influence of YKL-40 and TLR4 on NF-κB. There was a negative correlation between YKL-40 and TLR4 in group A, and the difference was statistically significant (P less than 0.05) while there was no relationship between YKL-40 and TLR4 expression in group B. The level of YKL-40 protein in group A was higher than that in group B, which was statistically significant (P less than 0.05). YKL-40 and TLR4 were positively correlated in group A while there was no correlation between YKL- 40 and TLR4 expression in group B. The expression of YKL-40, TLR4 and NF-κB in chronic sinusitis patients with nasal polyps was high. In addition, there was a negative correlation between YKL-40 and TLR4 expression in chronic sinusitis patients with nasal polyps. YKL-40 and TLR4 interacted with each other to activate NF-κB and promote disease progression.

Herein, we investigated the clinical value of concurrent radiochemotherapy for patients with advanced cervical cancer and its effects on adverse clinical symptoms. Forty patients with cervical cancer were recruited from January 2011 to January 2014 for this study. Participants were randomly allocated into a test or control group, with 20 patients in each group. Patients in the test group were treated with concurrent radiochemotherapy, whereas patients in the control group received only traditional radiotherapy. At the end of the observation period, clinical efficacy in the two groups was compared. Patients were followed up for 2 years, and the rates of recurrence, survival, and complications were compared; ultrasonographic findings before and after radiotherapy were also correlated. Patients in the test group who received concurrent radiochemotherapy showed significantly higher clinical efficacy than the control group at the end of treatment cycles. After 2 years of follow-up, the rates of recurrence, mortality, and complications were all significantly lower in the test group than in the control group (P < 0.05). Comparison of ultrasonographic findings before and after radiochemotherapy showed that the size of the tumor was significantly smaller in patients after concurrent radiochemotherapy. Compared with traditional radiotherapy, concurrent radiochemotherapy significantly improved clinical outcomes in patients with advanced cervical cancer. Concurrent radiochemotherapy also enhanced the rate of survival and decreased the rate of relapse, with enhanced clinical safety and no significant side effects. Thus, concurrent radiochemotherapy can be more broadly applied in the treatment of advanced cervical cancer.

To determine the cytotoxic effect of lymphocytes activated by melanoma-associated antigen 3 (MAGE-3)-sensitized dendritic cells (DCs) on BIU-87 tumor cells, and to evaluate the possibility of MAGE-3-peptide-pulsed DCs as a vaccine in bladder cancer immunotherapy, the proliferation of T cells and the activity of cytotoxic T lymphocytes (CTLs) were examined by the MTT method. CTLs were induced by MAGE-3-sensitized DCs, or by ovalbumin (OVA) peptide and non-sensitized DCs as controls, respectively. The results indicated that MAGE-3-sensitized DCs have the ability to promote the proliferation of T cells as well as the cytotoxic activity of CTLs on bladder cancer cells in comparison with OVA peptide and non-sensitized DCs. In other words, DCs sensitized by the MAGE-3 antigen peptide could obviously upregulate the proliferation of T cells, which resulted in the growth inhibition of bladder cancer BIU-87 cells. In addition, MAGE-3-sensitized DCs played an important role in inhibiting the growth of human BIU-87 tumor xenografts in nude mice.

Supplementary dietary plant oils have the potential to alter milk fatty acid composition in ruminants as a result of changes in the amount and kind of fatty acid precursors. We hypothesized that linseed oil in combination with malate (a key propionate precursor in the rumen) would increase ∆9 unsaturated fatty acids and specific gene expression in somatic cells and mammary glands of lactating goats. Twelve lactating goats were used in a 3 × 3 Latin square design. Treatments included the basal diet (CON), the CON plus 4% linseed oil (LO), and the CON plus 4% linseed oil and 2% -malate (LOM). Relative to CON, the LO and LOM supplements increased the daily intake of palmitic (16:0), stearic (18:0), oleic (18:1-9), linoleic (18:2-6), α-linolenic (18:3-3), and γ-linolenic acids (18:2-6); α-linolenic acid intake was increased over 9-fold, from 6.77 to over 51 g/d (0.02). The LO and LOM supplements increased daily milk yield, milk fat yield, and milk fat percentage (0.05). The LOM supplement also increased milk lactose percentage and daily yield ( = 0.03). Both the LO and LOM supplements increased plasma glucose and total cholesterol and decreased plasma β-hydroxbutyrate concentrations ( = 0.03). The LO and LOM supplements increased concentrations of stearic acid; -vaccenic acid (TVA; 18:1-11); -9, -11 CLA; -10 -12 CLA; and α-linolenic acid in rumen fluid and increased the concentrations of oleic acid; TVA; -9, -11 CLA; -10, -12 CLA; and α-linolenic acid in plasma lipids and milk fat (0.05). Conversely, the LO and LOM supplements decreased short- and medium-chain SFA, including lauric (12:0), myristic (14:0), and palmitic acids, in plasma and milk fat (0.05). Relative mRNA levels for and () gene expression were increased in somatic cells and mammary gland tissue by LO and LOM (0.05). We conclude that the higher intake and ruminal production of stearic acid promoted SCD gene expression in somatic cells and mammary tissue. Furthermore, milk somatic cells are a suitable substitute for documenting treatment effects of dietary oils on gene expression in goat mammary tissue.

The aim of the present study was to explore the role of selenoprotein P (SePP) in the etiology of the endemic sudden cardiac death in Yunnan, China. The levels of SePP of 124 subjects and glutathione peroxidase (GPx) of 119 subjects were measured. The subjects were from the old and new endemic areas and non-endemic areas. The levels of SePP and GPx of the subjects of the old endemic area were significantly higher than those of the subjects of the new endemic area and the non-endemic areas, respectively. The Pearson's correlation among SePP, GPx, and the number of the incident cases of the disease were statistically significant. These correlations show that there is an inverse relationship among the number of patients and the levels of SePP (r = - 0.9800, P = 0.0200) and GPx (r = - 0.961, P = 0.009). The results show that selenium deficiency might play an important role in the incidence of the disease.

To investigate the expression of autophagy-related gene Beclin1 and P62 in nasal polyps and its relationship with the pathogenesis of this disease.The specimens were divided into two groups: nasal polyp tissue(n=50) and normal inferior turbinate mucosa(n=20). The general morphology was detected with hematoxylin-eosin(HE) staining, the expression of Beclin1 and P62 was examined with immunohistochemistry(IHC) and real-time fluorescent quantitative reverse transcription-polymerase chain reaction ( RT-PCR). SPSS 20.0 software was used to analyze the data.Protein level: The expression of Beclin1 in nasal polyp tissue was lower than inferior turbinate mucosa(U=-13.36, P0.01), in contrast, P62 in experimental group was higher than control group(U=12.99 , P0.01). mRNA level: The relative quantity of Beclin1 and LC3B expressions in nasal polyp were 0.46±0.17 and 0.46±0.11, which was lower than those in turbinate mucosa 1.11±0.47 and 0.96±0.25.The differences were significant(t value was -4.61, -4.61, both P0.01). But the relative quantity of P62 expression in nasal polyp was 2.19±0.44, which was higher than that in turbinate mucosa (1.05±0.33). The difference was all significant(t=6.16, P0.01).Compared with control group, the expression of Beclin1 was deficient and P62 was much more. Autophagy was deficient in nasal polyps, which might be in connection with the pathogenesis of the disease.

Biodetoxification of mycotoxins is a novel strategy to control mycotoxicoses in animals. Bacillus sp. LS100, which transforms deoxynivalenol (DON) to a less toxic chemical de-epoxy DON (DOM-1), was evaluated for its efficacy in reducing the adverse effects of DON on swine growth performance. A feeding trial was conducted in growing pigs with four treatments: (1) corn meal without detectable DON served as control (Non-toxic Corn); (2) Fusarium-infected corn giving a toxic diet containing 5 µg DON g(-1) (Toxic Corn); (3) Toxic Corn detoxified with Bacillus sp. LS100 giving a detoxified diet containing 5 µg DOM-1 g(-1) (LS100-De-toxic Corn); (4) Non-toxic Corn treated with Bacillus sp. LS100 serving as bacterial control (LS100-Non-toxic Corn). During 9 days of exposure to the treatments, pigs on Toxic Corn showed a significant reduction in daily feed consumption, daily weight gain and feed efficiency by 29, 48 and 29%, respectively, compared to pigs on Non-toxic Corn. These parameters of the pigs fed LS100-De-toxic Corn diet were 45, 82 and 32% greater, respectively, than those of pigs fed Toxic Corn diet, and were similar to those pigs fed Non-toxic Corn and LS100-Non-toxic Corn diets. There were no significant differences between the treatments of LS100-Non-toxic Corn and Non-toxic Corn diets, implying that the bacterial isolate might not have significantly affected nutrition and palatability of the feed or had negative effects on the pig's feeding performance. The results have proved that microbial detoxification of DON in contaminated feed can eliminate negative effects of the mycotoxin, and the pre-feeding detoxification approach may be applied in the livestock industry.

International audience; Antimicrobial resistance is a global challenge that impacts both human and veterinary health care. The resilience of microbes is reflected in their ability to adapt and survive in spite of our best efforts to constrain their infectious capabilities. As science advances, many of the mechanisms for microbial survival and resistance element transfer have been identified. During the 2012 meeting of Antimicrobial Agents in Veterinary Medicine (AAVM), experts provided insights on such issues as use vs. resistance, the available tools for supporting appropriate drug use, the importance of meeting the therapeutic needs within the domestic animal health care, and the requirements associated with food safety and food security. This report aims to provide a summary of the presentations and discussions occurring during the 2012 AAVM with the goal of stimulating future discussions and enhancing the opportunity to establish creative and sustainable solutions that will guarantee the availability of an effective therapeutic arsenal for veterinary species.

Local tissue concentrations of glucocorticoids are modulated by the enzyme 11 β -hydroxysteroid dehydrogenase which interconverts cortisol and the inactive glucocorticoid cortisone in man, and corticosterone and 11-dehydrocorticosterone in rodents. The type I isoform (11 β -HSD1) is a bidirectional enzyme but acts predominantly as a oxidoreductase to form the active glucocorticoids cortisol or corticosterone, while the type II enzyme (11 β -HSD2) acts unidirectionally producing inactive 11-keto metabolites. There are no known clinical conditions associated with 11 β -HSD1 deficiency, but gene deletion experiments in the mouse indicate that this enzyme is important both for the maintenance of normal serum glucocorticoid levels, and in the activation of key hepatic gluconeogenic enzymes. Other important sites of action include omental fat, the ovary, brain and vasculature. Congenital defects in the 11 β -HSD2 enzyme have been shown to account for the syndrome of apparent mineralocorticoid excess (AME), a low renin severe form of hypertension resulting from the overstimulation of the non-selective mineralocorticoid receptor by cortisol in the distal tubule of the kidney. Inactivation of the 11 β -HSD2 gene in mice results in a phenotype with similar features to AME. In addition, these mice show high neonatal mortality associated with marked colonic distention, and remarkable hypertrophy and hyperplasia of the distal tubule epithelia. 11 β -HSD2 also plays an important role in decreasing the exposure of the fetus to the high levels of maternal glucocorticoids. Recent work suggests a role for 11 β -HSD2 in non-mineralocorticoid target tissues where it would modulate glucocorticoid access to the glucocorticoid receptor, in invasive breast cancer and as a mechanism providing ligand for the putative 11-dehydrocorticosterone receptor. While previous homologies between members of the SCAD superfamily have been of the order of 20–30% phylogenetic analysis of a new branch of retinol dehydrogenases indicates identities of >60% and overlapping substrate specificities. The availability of crystal structures of family members has allowed the mapping of conserved 11 β -HSD domains A–D to a cleft in the protein structure (cofactor binding domain), two parallel β -sheets, and an α -helix (active site), respectively.

Severe low-renin hypertension has few known causes. Apparent mineralocorticoid excess (AME) is a genetic disorder that results in severe juvenile low-renin hypertension, hyporeninemia, hypoaldosteronemia, hypokalemic alkalosis, low birth weight, failure to thrive, poor growth, and in many cases nephrocalcinosis. In 1995, it was shown that mutations in the gene ( HSD11B2 ) encoding the 11β-hydroxysteroid dehydrogenase type 2 enzyme (11β-HSD2) cause AME. Typical patients with AME have defective 11β-HSD2 activity, as evidenced by an abnormal ratio of cortisol to cortisone metabolites and by an exceedingly diminished ability to convert [11- 3 H]cortisol to cortisone. Recently, we have studied an unusual patient with mild low-renin hypertension and a homozygous mutation in the HSD11B2 gene. The patient came from an inbred Mennonite family, and though the mutation identified her as a patient with AME, she did not demonstrate the typical features of AME. Biochemical analysis in this patient revealed a moderately elevated cortisol to cortisone metabolite ratio. The conversion of cortisol to cortisone was 58% compared with 0–6% in typical patients with AME whereas the normal conversion is 90–95%. Molecular analysis of the HSD11B2 gene of this patient showed a homozygous C→T transition in the second nucleotide of codon 227, resulting in a substitution of proline with leucine (P227L). The parents and sibs were heterozygous for this mutation. In vitro expression studies showed an increase in the K m (300 nM) over normal (54 nM). Because ≈40% of patients with essential hypertension demonstrate low renin, we suggest that such patients should undergo genetic analysis of the HSD11B2 gene.

Background Apparent mineralocorticoid excess (AME) is a cause of low-renin, low-aldosterone hypertension in which cortisol acts as a mineralocorticoid. The condition reflects an inability to inactivate cortisol to cortisone due to defective activity of the type 2 isozyme of 11β-hydroxysteroid dehydrogenase (11β-HSD2). Homozygous mutations in 11β-HSD2 gene in patients with AME have been described. A 7-year-old Brazilian girl had previously been found to have AME. Her father recently presented with mineralocorticoid hypertension at age 38 years. Objective To describe the clinical details, to perform steroid analyses and to assess the molecular basis for the hypertension in this kindred. Methods The 11βHSD2 gene was amplified from genomic DNA by the polymerase chain reaction and sequenced by direct chain-termination sequencing on an automatic DNA sequencer. The sequencing results were validated by restriction-site polymorphism. The mutant 11β-HSD2 protein was expressed in Chinese hamster ovary polyoma cells and enzymatic activity was assessed by metabolizing cortisol in vitro. Results Sequence analysis of genomic DNA revealed a novel C1061T point mutation in exon V of the human 11β-HSD2 gene, resulting in an amino acid substitution of alanine by valine at codon 328 of the enzyme protein (A328V). Expression studies confirmed that the mutant protein was devoid of 11β-HSD2 activity. A Hhal restriction-site polymorphism confirmed that the proband was homozygous for the mutation whereas both parents were heterozygotes. The father of the proband had hypertension, a normal serum potassium level, suppressed plasma renin activity and plasma aldosterone level and a moderately elevated urinary cortisol: cortisone metabolite ratio. Conclusions AME in this kindred is caused by a novel mutation in the 11β-HSD2 gene. Detection of hypokalaemia, at least in this kindred, is an insensitive screening test for mineralocorticoid-based hypertension. In contrast to results from previously investigated kindreds, we have demonstrated that this kindred has an abnormal phenotype in the heterozygote state. Further studies are now required in order to evaluate the role of 11β-HSD2 activity in the pathophysiology of 'essential' hypertension.

The 11beta-hydroxysteroid dehydrogenase type II enzyme (11betaHSD2) endows specificity on the mineralocorticoid receptor by metabolising glucocorticoids. Sequence comparisons with other microsomal proteins showed the strongly preferred topology of a lumenal pentapeptide followed by three transmembrane helices with residues beyond Ala73 on the cytoplasmic side of the membrane, suggesting that 11betaHSD2 is anchored to the endoplasmic reticulum by the N-terminal region. However, deletion of the N-terminus (11betaHSD2 deltaN) and expression of the construct in mammalian cells showed that the enzyme remained bound to the microsomal fraction, indicating that other regions are also involved in membrane anchoring. Crosslinking studies and nonreducing SDS-PAGE demonstrated that 11betaHSD2 is a non-covalently linked dimer. Deletion of the non-conserved C-terminal region (11betaHSD2 deltaC) resulted in an enzyme with a Km of 215 nM for cortisol in whole cell assays, while 11betaHSD2 and 11betaHSD2 deltaN displayed a Km of 62 and 74 nM, respectively. In homogenates 11betaHSD2 and 11betaHSD2 deltaC displayed maximal activity at 140 mM NaCl or KCl, but showed a marked decrease in enzyme activity with increasing salt. 11BetaHSD2 was more stable than 11betaHSD2 deltaC in the presence of NaSCN, suggesting that the C-terminal region plays a role in enzyme stability. There was no detectable activity in homogenates containing 11betaHSD2 deltaN, while 11betaHSD2 deltaC and 11betaHSD2 displayed a Km of 135 and 46 nM, respectively. Although 11betaHSD2 is conventionally considered a unidirectional dehydrogenase all constructs converted 11-dehydrodexamethasone to dexamethasone in whole cell assays, providing an explanation for the potency of the synthetic glucocorticoid in the face of a powerful inactivator of natural glucocorticoids.

We examined the underlying neural-endocrine mechanisms of asthma associated with respiratory syncytial virus infection. Thirty Sprague-Dawley rats were randomly divided into control group, respiratory syncytial virus (RSV) group, and anti-nerve growth factor (NGF) IgG group. An RSV infection model was established by nasal drip once a week. In the anti-NGF antibody intervention group, each rat was given an intraperitoneal injection of anti-NGF IgG 3 h before RSV infection. Optical microscopy and transmission electron microscopy were used to observe the structural changes in adrenal medulla cells. Changes in adrenaline and norepinephrine in serum were detected by ELISA. NGF expression was assayed by immunohistochemistry. Expression differences in synaptophysin mRNA were detected by RT-PCR. Transmission electron microscopy displayed widened adrenal medulla intercellular spaces, reduced chromaffin particle concentration, and increased mitochondria in the RSV infection group. At the same time, NGF expression was increased in the RSV infection group significantly. In addition, the adrenaline concentration was significantly decreased compared with the control and anti-NGF antibody groups. Synaptophysin mRNA expression was significantly increased in the RSV infection and anti-NGF antibody groups. However, compared with the RSV infection group, synaptophysin mRNA expression was significantly decreased in the anti-NGF antibody group. We conclude that RSV infection could induce adrenal medulla cell differentiation to nerve cells by over-expression of NGF, resulting in the decreased endocrine function found in asthma progression.

Kava is known for its recreational, ceremonial and medicinal use in the Pacific. The aqueous non-alcoholic drink of kava rhizome produces intoxicating, relaxing and soothing effects. While kava's medicinal effects receive worldwide recognition, kava-containing products came under scrutiny after over 100 reports of spontaneous adverse hepatic effects. Many mechanisms have been postulated to explain the unexpected toxicity, one being pharmacokinetic interactions between kavalactones and co-administered drugs involving cytochrome P450 enzyme system. Alcohol is often co-injested in kava hepatotoxicity cases. This review evaluates the possible hepatotoxicity mechanisms involving alcohol and kava.

To establish the quality standards for fuyanke(FYK) granule.The phellodendron Chinese schneid, polygonum cuspidatum, sieb. et Zucc and glycyrrihza uralensis fisch in FYK granule were identified by thin-layer chromatography(TLC). The contents of berberine were determined by thin-layer chromatographic scanning(TLCs).The average recovery rate was 97.86%, and the relative standard deviation 1.96%.The method is simple, accurate and reliable.

We describe a new member of the 17beta-hydroxysteroid dehydrogenase group of enzymes. Human Pan1b displays greatest activity with 5alpha-androstan-3alpha,17beta-diol (3alpha-Diol) as substrate, suggesting that it may be important in androgen metabolism. Enzymic activity was non-saturable with 3alpha-Diol but saturable with retinoids, although retinoids were not metabolized. Immunohistochemical studies on 10% formalin fixed and paraffin embedded sections of human tissues showed that Pan1b was present in acini and ciliated epithelia of the lung. In the fetus immuno reactivity was present in ciliated epithelia throughout gestation and staining appeared to be stronger in the second half of pregnancy. Pan1b was also expressed in the nonpigmented epithelium of the ciliary body, and in adrenocortical tumor cells. Although 3alpha-Diol is generally considered a degradation product of androgen metabolism it could have its own biological function. Pan1b may be an important modulator of the endocrine, or intracrine activity of this steroid.

The 11beta-hydroxysteroid dehydrogenase type 2 enzyme (11betaHSD2) metabolizes glucocorticoids into their inactive 11-keto metabolites. Although the type 1 enzyme (11betaHSD1) displays both oxidative and reductive activity, to date 11betaHSD2 has been shown to have dehydrogenase activity only. In this study we compared both dehydrogenase and reductase characteristics of the cloned rat 11betaHSD1 and rat and human 11betaHSD2 for three different 11-hydroxysteroid substrates, cortisol (F), corticosterone (B), and dexamethasone (Dex), and the corresponding 11-keto metabolites, cortisone (E), 11-dehydrocorticosterone (A), and 11-dehydrodexamethasone (DH-Dex), respectively. In cell homogenates expressing either the rat or the human 11betaHSD2, the relative potency for the dehydrogenase reaction was BFDex. Although there was no reduction of A or E, DH-Dex was readily converted to Dex with an equilibrium far on the side of the 11-hydroxy metabolite. DH-Dex reduction in homogenates was inhibited by both glycyrrhetinic acid and carbenoxolone, with a 50% inhibition at 80 and 100 nM, respectively. In intact cells transfected with rat 11betaHSD1, the equilibrium was on the reductase side for all substrates. Dehydrogenation of B or F was more potent with rat 11betaHSD2 than with rat 11betaHSD1. There was no detectable 11betaHSD1 oxidation of Dex. These data indicate that both the cloned human and rat 11betaHSD2 reduce DH-Dex and do this more readily than they oxidize Dex. Thus, 11betaHSD2 seems also to be a bidirectional enzyme, although no reduction of the physiological compounds A and E was observed.

Mineralocorticoid action is facilitated by 11 beta-hydroxysteroid dehydrogenase type II (11 beta HSD2), which metabolizes glucocorticoids and allows aldosterone to bind to the nonselective mineralocorticoid receptor. We have recently demonstrated the presence of the 11 beta HSD2 protein in a wide range of human epithelia, suggesting that it is the sole isoform endowing specificity in man. In the present study we have used an immunopurified polyclonal antibody (RAH23) raised against a C-terminal peptide derived from the cloned rat 11 beta HSD2 protein to perform immunohistochemical and molecular analysis in rat tissues. In frozen sections of rat kidney, strong staining was seen with the RAH23 antibody in the distal tubule; weaker staining was observed in the thick ascending loop of Henle and the medullary and papillary collecting ducts. Punctate cortical staining was observed in the fetus at 20 days gestation and in 8-day-old rats, with a noticeable increase in the staining pattern at 16 days of age. The kidney did not attain the adult pattern of staining until 28 days of age. Epithelia of ileum and colon also stained with RAH23, as did excretory ducts of the submandibular gland. Intrahepatic and excretory bile ducts displayed strong immunoreactivity in the epithelial lining. Rat adrenal glands showed evidence of the 11 beta HSD2 antigen in the zona fasciculata and zona reticularis, but not in the zona glomerulosa or medulla. Western blot analysis with the RAH23 antibody revealed strong bands in the kidney, colon, adrenal gland, and submandibular gland at 40 kDa, colinear with the migration of the cloned 11 beta HSD2 enzyme. A band of medium intensity was also seen at this size in the pancreas, whereas a band of moderate intensity was seen in the bile duct, and weaker bands were noticed in the stomach, small intestine, and liver, with a diffuse band at 36-42 kDa in the prostate. Strong bands were seen in the pancreas and prostate at 78 kDa, with weaker signals in the colon, adrenal, stomach, and bile duct. A number of tissues also displayed multiple bands at about 30 kDa. Enzymatic assays on tissue homogenates showed extensive conversion of corticosterone to its 11-dehydro product in an NAD-dependent manner in the submandibular gland, adrenal gland, and kidney, but not in the pancreas or prostate. This study confirms the ubiquitous presence of 11 beta HSD2 in sodium-transporting epithelia, demonstrates the high level of 11 beta HSD2 protein and enzyme activity in the rat adrenal, and suggests a possible role for the enzyme in the biliary system. Further studies are required to determine the relevance of the various molecular species to the activity, latency, and processing of the enzyme.

The 11 beta-hydroxysteroid dehydrogenase type II enzyme (11 beta HSD2) converts cortisol into mineralocorticoid receptor inactive cortisone, thus preventing occupation of the non-selective mineralocorticoid receptor by glucocorticoids in the kidney. Mutations generating inactive enzymes have been described in the HSD11B2 gene in the congenital syndrome of apparent mineralocorticoid excess (AME), although proof of mutant protein synthesis was not provided. In the present study we have examined the metabolism of cortisol in mammalian cells transfected with plasmids expressing the wild type and mutant enzymes from three additional families of patients with mutations in the HSD11B2 gene. These studies revealed that the mutants were enzymatically inactive in intact mammalian cells expressing significant levels of both full length and truncated proteins. This is the first study to definitively show that point mutations in the HSD11B2 gene abolish 11 beta HSD2 enzymatic activity in the syndrome of AME.

Human Pro-Urokinase (Pro-UK) is expressed in CHO-DHFR- cells at high efficiency by co-transfecting the mouse dhfr gene and Pro-UK cDNA under the control of the SV40 late promoter. After gene co-amplification, the product level could reach 2-3 micrograms/10(6) cells/24 h in the presence of 5 x 10(-6) mol/L MTX, and the levels can be further raised to 3.5-4 micrograms/10(6) cells/24 h by PMA superinduction. The copy number of Pro-UK cDNA in the genomes of host cells is about 200-300 copies/cell. The Western blot analysis shows that the recombinant Pro-UK has similar molecular weight to its natural counterpart, and also the amidolytic activity of the product is determined by S-2444 assay.

Transcranial Doppler (TCD) was performed in 64 patients with subarachnoid hemorrhage (SAH) to detect the blood velocity in middle cerebral artery (MCA). Based on Poiseuille's principle, the increase of blood velocity was used as an indicator of cerebral vasospasm (CVS). It was found that 61.67% of the patients with SAH developed CVS, especially in the second week after SAH. The incidence of CVS in patients below 50 years old was higher than that in patients above 51 years old. The results showed that TCD is valuable in evaluation of CVS after SAH.

Expression vectors containing the pro-urokinase (pro-UK) cDNA (pSV2-proUK) and a dihydrofolate reductase cDNA (pSV2-dhfr or MMTV-dhfr) were cotransfected into CHO-dhfr- cells by the calcium phosphate precipitation technique. The dhfr+ transformants were selected by fibrinolytic agarose plate assay. Two colonies, named CLF-14 and CLF-8, exhibited significantly high expression levels of the biological activity of urokinase-type plasminogen activator (mu-Pa). They reached more than 24 IU/10(6) cells/48 h and 16 IU/10(6) cells/48 h, respectively. Examination of the cell supernatants for mu-Pa antigenicity using ELISA method also showed strong positive results, and the quantities of expression were about 0.14-0.22 micrograms/10(6) cells/48 h and 0.08-0.14 micrograms/10(6) cells/48 h, respectively. The mu-Pa secreted by stable transformed cells could be completely inhibited by UK anti-serum, but not by tissue-type plasminogen activator (t-PA) antiserum nor by normal rabbit serum.

Seven cases of ovarian sex cord tumor with annular tubules are reported in this paper. All lesions occurred in the young (6-25 years of age with a mean of 15 years) without Poutz-Jeghers syndrome. Four cases had estrogenic hormonal imbalance and abdominal mass. Gross examination revealed that the tumors were unilateral with smooth surface. Except one, all showed varying degrees of cystic degeneration on section. The sections appeared as roe-like granular substance in a pale yellowish brown shade in the parenchymal region. Microscopic examination showed single or complex annular tubules with prominent basement membrane-like material around the tubules. Lipid in the cytoplasm and granulosa cell tumor-like differentiation were observed in isolated area of some cases. Ultrastructurally, Charcot-Battcher filaments were found in 1 case, indicating differentiation towards Sertoli's cells. The malignant potential of this tumor had been elucidated in the literature.

Poly(A)+ RNA was isolated from 9 specimens of human primary hepatic carcinoma, 1 non-tumorous liver tissue adjacent to cancer and 1 normal liver tissue samples. The Oligo-dT cellulose-purified poly(A)+ RNAs were subjected to formaldehyde agarose gel electrophoresis, Northern transfer and hybridization with various oncogene probes. Two RNA species, 5.6 kb and 2.2 kb were identified by N-ras gene hybridization in 6 out of 9 mRNA samples from primary hepatic carcinoma specimen. N-ras specific mRNA was not detectable in mRNA samples from normal human liver and tumor surrounding cirrhotic tissue. No detectable hybridization of mRNA from hepatoma and normal liver with Ki-ras or Ha-ras was observed. As human N-ras gene has been identified in DNA of mouse transfectants transformed with PHC DNA, it strongly suggests that N-ras gene might be responsible for the transforming activity of part of cases of human liver cancer.

DNA was extracted from NIH 3T3 cells transformed with DNAs from human primary hepatic cancer (PHC) and Hepatoma 7402 cell line. The transformant DNA was analyzed by Southern transfer and hybridization with 32P-labeled probes of various oncogenes. The EcoRI 7.2 and 9.0 kb bands characteristic of human N-ras gene were identified in transformed NIH 3T3 cells derived both from PHC and 7402 DNA. The BamHI 6.6 kb band characteristic of human c-Ha-ras I was present only in 7402 transformants, but not in PHC transformants. Using 35S-methionine incorporation, immunoprecipitation with anti-p21 monoclonal antibodies, SDS-PAGE and autoradiography, it was demonstrated that p21 synthesis was remarkably enhanced in 7402 cells as well as in transformed cells derived from both 7402 and PHC DNA. Taking the data together, it strongly implies that N-ras is one of the transforming genes for human liver cancer.