[Reference values for carotid artery intima-media thickness among community adult dwellers in Shenzhen City].

Objective: To establish reference values for carotid intima-media thickness (CIMT) of adult dwellers in Shenzhen City. Methods: The study was conducted based on the Shenzhen heart failure epidemiological survey from 2021 to 2022. In this survey, residents aged 18 years and above in Shenzhen were selected by using a multi-stage stratified random sampling method. General information, cardiovascular disease (CVD) related behavior and carotid ultrasound examination and etc. were collected from the participants. People with CVD factors, a history of atherosclerotic cardiovascular disease, carotid plaque or having no carotid ultrasound examination results were excluded. The parameter regression model based on fractional polynomial was used to establish the reference values of CIMT by age and sex. Results: A total of 2 163 healthy individuals were enrolled in the final analysis, including 576 males (26.6%) and 1 587 females (73.4%). The fractional polynomial regression of the CIMT mean and standard deviation was obtained. For men, the regression was meanCIMT=0.324 7+0.006 9×age and SDCIMT=0.076 9+0.001 2×age. For women, the regression was meanCIMT=0.354 9+0.005 4×age and SDCIMT=0.041 6+0.002 0×age. Conclusion: The age and sex reference values for CIMT of adult people in Shenzhen established in this study could provide the latest reference standards for early screening of subclinical CVD.

[A retrospective controlled study of TACE-HAIC-targeted-immune quadruple therapy for intermediate and advanced-stage hepatocellular carcinoma].

Objective: To evaluate the efficacy and safety of transcatheter arterial chemoembolization (TACE)-hepatic arterial infusion chemotherapy (HAIC)-targeted-immune quadruple therapy in patients with intermediate and advanced-stage hepatocellular carcinoma (HCC). Methods: 101 patients with intermediate and advanced stage HCC were enrolled according to the inclusion and exclusion criteria, and then they were divided into a combination group and a control group. Patients in the combination group was treated with TACE-HAIC-targeted-immune quadruple therapy, while the control group was only treated with TACE therapy. The overall survival (OS), progression-free survival (PFS), and treatment-related adverse reactions were statistically analyzed in the two groups of patients. Statistical analysis was carried out by t-test, χ2 test, rank sum test, Kaplan-Meier curve, log-rank test, Cox regression (or proportional hazards model) analysis according to different data. Results: The tumor objective response rate and disease control rate as evaluated by mRECIST 1.1 criteria in the combination group were 80% and 94%, respectively, which were significantly higher than those in the control group, 41.2% (P<0.001) and 74.5% (P=0.007). The OS and PFS of the combination group were 15.6 months [95%CI 11.3-NA ] and 8.8 months [95%CI 6.9-12.0], respectively, which were significantly better than the control group at 6.1 months [95%CI 5.3-6.6] (P<0.001) and 3.2 months [95%CI 3.0-3.6] (P<0.001). Gastric ulcer incidence was significantly higher in the combination group (9/50, 18%) than that in the control group (2/51, 3.9%) (P=0.023). Conclusion TACE-HAIC-targeted-immune quadruple therapy is a more effective treatment mode for intermediate and advanced-stage HCC than TACE alone, and attention should be paid to the monitoring of target immune-related adverse reactions.

[Clinical features of community-acquired bloodstream infection due to Gram-negative bacilli in patients with liver cirrhosis].

Objective: o investigate the features of pathogenic bacteria for community-acquired bloodstream infection due to Gram-negative bacilli in patients with liver cirrhosis and optimal therapeutic strategy. Methods: A retrospective analysis was performed for the clinical data of patients with liver cirrhosis who were admitted to 302 Hospital of PLA due to community-acquired bloodstream infection from January 2010 to December 2015, and a statistical analysis was performed for their clinical features, pathogenic bacteria, and results of drug sensitivity test. The Pearson chi-square test was used for comparison of rates, and the Wilcoxon rank sum test was used for comparison of ranked data. Results: A total of 240 patients (including 178 male patients) with liver cirrhosis caused by various reasons were enrolled, with a mean age of 51.7 ± 11.1 years, an overall clinical remission rate of 80.42%, and an ineffective/mortality rate of 19.58%. The patients who used sensitive antibiotics within 12 hours after the onset of community-acquired bloodstream infection achieved a significantly higher improvement rate than those who used such drugs at more than 12 hours after onset (88.2% vs 58.1%, P < 0.001). The improvement rate achieved by the application of sensitive antibiotics at more than 12 hours after onset decreased with the increase in the Child-Pugh grade (P < 0.05). A total of 245 strains of Gram-negative bacilli were isolated, among which the six most common ones were 135 strains of Escherichia coli (55.1%), 62 strains of Klebsiella pneumoniae (25.3%), 16 strains of Aeromonas (6.5%), 4 strains of non-typhoidal Salmonella (1.6%), 3 strains of Enterobacter cloacae (1.2%), and 2 strains of Acinetobacter baumannii (0.8%). These Gram-negative bacilli had the highest sensitivity to meropenem (98.5%), followed by imipenem (97.9%), amikacin (97.5%), piperacillin/tazobactam (94.7%), cefmetazole (93.7%), and cefoperazone/sulbactam (93%). Different bacteria had different sensitivities to antibiotics. Conclusion: Once community-acquired bloodstream infection occurs in patients with liver cirrhosis, highly sensitive antibiotics should be used as early as possible. Cefoperazone/sulbactam, piperacillin/tazobactam, imipenem, and meropenem can be used as first-line empirical antibiotics, and drug combination should be considered when necessary.

[Simultaneous determination of various volatile organic compounds in urine by headspace GC-MS].

Objective: To establish a method for determination of acetone, dichloromethane, hexane, 1, 1, 1-trichloroethane, 1, 2-dichloroethane, benzene, toluene, ethylbenzene etc organic compounds in urine by headspace gas chromatography-mass spectrometry (GC-MS) . Methods: Headspace gases of urine samples were injected into GC and determined by mass. Results: Determination of urine components were in a good linear range in their concentration range of this method. The correlation coefficients were between 0.996 and 1.000 with the detection limits between 0.1 µg/L and 4.5 µg/L, the precisions were between 1.3% and 4.6%, the recovery rates were between 86.2% and 97.4%. Conclusion: This method has the advantages of low detection limits, high accuracy, high precision and simple pretreatment, which is suitable for the determination of the content of various volatile organic compounds in urine.

Clinical and immunological analysis of measles patients admitted to a Beijing hospital in 2014 during an outbreak in China.

At the end of 2013, China reported a countrywide outbreak of measles. From January to May 2014, we investigated the clinical and immunological features of the cases of the outbreak admitted to our hospital. In this study, all 112 inpatients with clinically diagnosed measles were recruited from the 302 Military Hospital of China. The virus was isolated from throat swabs from these patients, and cytokine profiles were examined. By detecting the measles virus of 30 of the 112 patients, we found that this measles outbreak was of the H1 genotype, which is the major strain in China. The rates of complications, specifically pneumonia and liver injury, differed significantly in patients aged 18 years: pneumonia was more common in children, while liver injury was more common in adults. Pneumonia was a significant independent risk factor affecting measles duration. Compared to healthy subjects, measles patients had fewer CD4+IL-17+, CD4+IFN-γ +, and CD8+IFN-γ + cells in both the acute and recovery phases. In contrast, measles patients in the acute phase had more CD8+IL-22+ cells than those in recovery or healthy subjects. We recommend that future studies focus on the age-related distribution of pneumonia and liver injury as measles-related complications as well as the association between immunological markers and measles prognosis.

[Determination of tert-butyl alcohol in the air of the workplace by solvent desorption gas chromatography].

Objective: To establish solvent desorption gas chromatographic method for determination of tert-butyl alcohol in the air of the workplace. Methods: After tert-butyl alcohol in the air of the workplace collected with activated carbon tube and desorbed with 2% 2-propanol in CS2, and then separated with DB-FFAP capillary column and detected with flame ionization detector. Results: The linearity ranges were 0.6~2 264.0 mg/L. The limit of quantification was 0.6 mg/L. The determination has a good reproducibility. The intraassay and interassay precisions were 2.8%~3.2% and 3.8%~5.7%. The desorption efficiencies were 93.9%~98.1%. Absorption efficiencies were 95.8%~100.0%. The breakthrough volume was above 7.1 mg in 100mg activated carbon. The samples in activated carbon tube could be stored for at least 14 days at ambient temperature. Conclusion: The method is feasible for determination of tert-butyl alcohol in the air of the workplace.

Alarming incidence of genital mycoplasmas among HIV-1-infected MSM in Jiangsu, China.

Males who have sex with men (MSM) are considered at high risk of blood-borne and sexually transmitted infections (STIs), mainly due to the practice of unsafe sex, often combined with drug use and needle-sharing. A cross-sectional study was designed for the detection of genital mycoplasmas during the period from March 2009 to May 2010 in Jiangsu province. This work was approved by the Research ethics Committee of Jiangsu Centers for Diseases Prevention and Control (CDC), and written consent was obtained from all participants. In total, 243 human immunodeficiency virus-1 (HIV-1)-infected MSM were screened in this study. Over half of them reported a history of sexual activity with females (65.0 %), and 26.3 % reported a history of sexually transmitted diseases (STDs) other than HIV. 44.0 % of patients were in the first 2 years of their HIV infection, and 72.4 % were still in HIV progression. Of the 243 analyzed samples, all were positive for at least one kind of mycoplasma. The infection rates of Mycoplasma genitalium, M. fermentans, M. penetrans, and M. pirum were 25.5, 9.9, 2.5, and 18.5 %, respectively. The M. genitalium infection was associated with a history of sexual activity with females, and those who had sex with females showed higher infection rates. Six M. penetrans-positive patients were still in HIV infection progression and did not receive highly active antiretroviral therapy (HAART). Men who perform this particular behavior are at higher risk of Mycoplasma infections. Further molecular and epidemiological cohort studies with larger populations are needed in order to identify the role of Mycoplasma infections in HIV-1-infected MSM.

Comparative analysis of bilirubin in correlation to albumin between nephrotic syndrome patients and postoperative gastroparesis syndrome patients.

This study aimed to disclose the potential causality of low bilirubin in patients with nephrotic syndrome (NS). Correlation analysis was carried out on total bilirubin (TBIL) to serum albumin (ALB), urine protein (Upr), and urinary microalbumin/creatinine (Umalb/cr) for three groups in a case-control study. P < 0.001 was observed for TBIL, ALB, Umalb/cr, and Upr between the NS and chronic nephritis (CN) groups, and P values of 0.0001, 1.000, 0.0001, and 0.0001 were observed for TBIL, ALB, Umalb/cr, and Upr, respectively, between the postoperative gastroparesis (PGS) and CN groups. The values of r and P in correlation to TBIL were 0.549 and 0.000 for ALB, -0.405 and 0.000 for Umalb/cr, and -0.448 and 0.000 for Upr in the NS group; -0.007 and 0.959 for ALB, 0.213 and 0.091 for Umalb/cr, and -0.082 and 0.519 for Upr in the PGS group; and 0.509 and 0.000 for ALB, -0.431 and 0.000 for Umalb/cr, and -0.362 and 0.002 for Upr in the CN group. A probable causality is implied between the low level of blood bilirubin and its loss in urine in NS patients. This conclusion may provide a theoretical basis for the feasibility of therapies against oxidative stress in NS patients.

Clinical efficacy and adverse reactions analysis of PD-1/PD-L1 inhibitors in advanced esophageal squamous cell carcinoma.

OBJECTIVE: The aim of this study was to investigate the clinical efficacy and associated adverse reactions of Programmed Death Receptor-1 (PD-1)/PD-L1 inhibitors in the management of patients with advanced esophageal squamous cell carcinoma (ESCC). PATIENTS AND METHODS: In this retrospective study, 54 patients with advanced ESCC treated with PD-1/PD-L1 inhibitors in our hospital from January 2021 to January 2023 were identified as the research subjects. Using propensity score matching at a 1:1 ratio, patients only receiving chemotherapy were recruited as controls. The clinical effectiveness of PD-1/PD-L1 was evaluated by comparing the objective response rate (ORR) and disease control rate (DCR). Progression-free survival (PFS) and overall survival (OS) were analyzed for treatment outcome assessments. Adverse events (AEs) between the two groups were recorded and compared. RESULTS: Patients treated with PD-1/PD-L1 inhibitors had a higher rate of ORR (33.33%) and disease control (85.19%), compared to controls with an objective response rate of 20.37% and a disease control rate of 59.26%. The two groups showed similar ORR results, while the incorporation of PD-1/PD-L1 inhibitors resulted in significantly increased DCR when compared to the controls. The median OS was 22 months (95% CI: 1,629 months) for the control group and 31 months (95% CI: 28NA) for the study group, suggesting OS benefits offered by PD-1/PD-L1 inhibitor treatment (HR=0.479, 95% CI: 0.284, 0.809). The median PFS was 15 months (95% CI: 1,223 months) for the control group and 23 months (95% CI: 1,926) for the study group, indicating more PFS benefits provided by PD-1/PD-L1 inhibitors (HR=0.662, 95% CI: 0.436, 1.005). Adverse events and their severity were recorded during patient follow-up, and no grade 5 adverse events were reported in either group. The incidence of grade 3 or higher adverse events between the two groups was similar, while PD-1/PD-L1 inhibitors appeared to significantly reduce the incidence of gastrointestinal reactions in patients. CONCLUSIONS: PD-1/PD-L1 inhibitors integrated with chemotherapy provide significant benefits in the management of patients with advanced ESCC without increasing adverse events.

Separation and identification of Musa acuminate Colla (banana) leaf proteins by two-dimensional gel electrophoresis and mass spectrometry.

To establish a proteomic reference map of Musa acuminate Colla (banana) leaf, we separated and identified leaf proteins using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and mass spectrometry (MS). Tryptic digests of 44 spots were subjected to peptide mass fingerprinting (PMF) by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS. Three spots that were not identified by MALDI-TOF MS analysis were identified by searching against the NCBInr, SwissProt, and expressed sequence tag (EST) databases. We identified 41 unique proteins. The majority of the identified leaf proteins were found to be involved in energy metabolism. The results indicate that 2D-PAGE is a sensitive and powerful technique for the separation and identification of Musa leaf proteins. A summary of the identified proteins and their putative functions is discussed.

Relationship between NogoA/NgR1/RhoA signaling pathway and the apoptosis of cerebral neurons after cerebral infarction in rats.

The article "Relationship between NogoA/NgR1/RhoA signaling pathway and the apoptosis of cerebral neurons after cerebral infarction in rats, by Y.-X. Xie, M. Zhang, C.-R. Zhang, F. Chen, published in Eur Rev Med Pharmacol Sci 2020; 24 (1): 295-303-DOI: 10.26355/eurrev_202001_19924-PMID: 31957843" has been retracted by the authors due to some inaccuracies. The authors state that they recently conducted animal experiments again to verify the state of apoptosis in the brain tissue of the CI + NogoA KO group. However, it was found that the phenomenon of apoptosis in the tissue was not significantly inhibited, and the same result was subsequently confirmed by RT-PCR. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/19924.

Isolation and characterization of nucleotide-binding site and C-terminal leucine-rich repeat-resistance gene candidates in bananas.

Commercial banana varieties are highly susceptible to fungal pathogens, as well as bacterial pathogens, nematodes, viruses, and insect pests. The largest known family of plant resistance genes encodes proteins with nucleotide-binding site (NBS) and C-terminal leucine-rich repeat (LRR) domains. Conserved motifs in such genes in diverse plant species offer a means for the isolation of candidate genes in banana that may be involved in plant defense. Six degenerate PCR primers were designed to target NBS and additional domains were tested on commercial banana species Musa acuminata subsp malaccensis and the Musa AAB Group propagated in vitro and plants maintained in a greenhouse. Total DNA was isolated by a modified CTAB extraction technique. Four resistance gene analogs were amplified and deposited in GenBank and assigned numbers HQ199833-HQ199836. The predicted amino acid sequences compared to the amino acid sequences of known resistance genes (MRGL1, MRGL2, MRGL3, and MRGL4) revealed significant sequence similarity. The presence of consensus domains, namely kinase-1a, kinase-2 and hydrophobic domain, provided evidence that the cloned sequences belong to the typical non-Toll/interleukin-1 receptor-like domain NBS-LRR gene family.

Relationship between NogoA/NgR1/RhoA signaling pathway and the apoptosis of cerebral neurons after cerebral infarction in rats.

OBJECTIVE: The aim of this study was to investigate the effect and the mechanism of the NogoA/NgR1/RhoA signaling pathway on the apoptosis of neurons in cerebral infarction (CI) rats. Our findings might provide references for clinical prevention and treatment of CI. MATERIALS AND METHODS: A total of 60 adult male Wistar rats were randomly divided into 3 groups, including: Sham operation group (Sham group), CI group, and CI + NogoA gene knockout group (CI + NogoA KO group) using a random number table. The model of CI was successfully constructed using suture method in rats of CI group and CI + NogoA KO group. Only blood vessels were exposed in Sham group. At 2 days after CI operation, the rats were killed, and brain tissues were collected. Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and Western blotting were used to detect the messenger ribonucleic acid (mRNA) and protein expression levels of NogoA/NgR1/RhoA in brain lesion tissues of rats in the three groups, respectively. Subsequently, the pathological damage of brain tissues was detected via hematoxylin and eosin (H&E) staining. TTC staining was carried out to evaluate the infarction area in each group. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was conducted to measure the apoptosis level of neurons in brain tissues of rats in each group. Additionally, the level of Nissl's body in brain tissues of each group was examined by Nissl staining. Furthermore, the expression level of the platelet-derived growth factor (PDGF) in brain tissues of rats in the three groups was measured via immunohistochemistry. RESULTS: The mRNA and protein expression levels of the NogoA/NgR1/RhoA signaling pathway in brain tissues of rats in CI group increased significantly (p<0.05). NogoA KO could significantly reduce the infarction area of brain tissues in rats (p<0.05). H&E staining and Nissl's body staining revealed that neurons in the brain tissues of rats showed evident edema and disordered arrangement after CI. Meanwhile, the number of Nissl's body was remarkably reduced. However, after KO of NogoA, brain tissue damage was significantly alleviated in rats, and the number of Nissl's body increased remarkably at the same time (p<0.05). According to TUNEL staining results, inhibiting NogoA could notably reverse CI-induced apoptosis of neurons in brain tissues of rats (p<0.05). Immunohistochemical staining results demonstrated that the expression of PDGF in brain tissues of rats in CI group decreased markedly, whereas was significantly elevated in rats of CI + NogoA KO group (p<0.05). CONCLUSIONS: The expression of the NogoA/NgR1/RhoA signaling pathway was significantly elevated in brain tissues of CI rats. Furthermore, suppressing the NogoA/NgR1/RhoA signaling pathway could reduce CI-induced apoptosis of neurons in rats.

Influence of LincRNA-p21 on acute lung injury in sepsis.

OBJECTIVE: Acute lung injury (ALI) is one of the most serious complications of sepsis and remains refractory. It is of great significance to discuss the pathogenesis of acute lung injury in sepsis and look for more effective drugs for treatment. The purpose of this study was to investigate the role of LincRNA-p21 on acute lung injury in sepsis. MATERIALS AND METHODS: Lung histology was detected by HE staining to evaluate sepsis-induced ALI model in rats. The miRNA expression of LincRNA-p21 in septic model in vivo and in vitro was detected by RT-qPCR. Cell apoptosis, inflammatory responses and oxidative stress were detected to uncover the influence of LincRNA-p21 on LPS-induced septic model in vitro. RESULTS: The expression of LincRNA-p21 was significantly increased in septic model in vivo and in vitro. Cell apoptosis, inflammatory responses and oxidative stress were alleviated by LincRNA-p21 interference in LPS-treated BEAS-2B cells. CONCLUSIONS: All the results in the current study proved that LincRNA-p21 interference could alleviate the acute lung injury in septic model. It raised the conclusion that LincRNA-p21 may act as a novel regulator in the pathological process and a potential therapeutic target in sepsis-induced ALI.

[The effects of postauricular injection of methylprednisolone on medium-high frequency sudden hearing loss].

Objective:To study the effects of postauricular injection of methylprednisolone on medium-high frequency sudden hearing loss.Method:The data of 45 patients with invalid treatment who were diagnosed as medium-high frequency sudden hearing loss were retrospectively studied. They were divided into postauricular injection group and non-postauricular injection group. The treatment efficiency and hearing improvement at each frequency were compared between the two groups.Result:The improvements of hearing threshold in the non-postauricular injection group were(6.30±5.00)dB HL,(3.80±5.52)dB HL,(5.35±5.50)dB HL,(15.60±11.84)dB HL,(19.60±13.46)dB HL,(15.40±12.90)dB HL at 250, 500, 1000, 2000, 4000 and 8000Hz, respectively. The hearing improvements of the postauricular injection group were (5.35±6.22)dB HL,(2.50±3.00)dB HL,(4.65±6.60)dB HL,(23.75±10.75)dB HL,(25.75±11.73)dB HL,(30.50±14.50)dB HL at 250, 500,1000,2000,4000 and 8000Hz, respectively. There were significant differences between the two groups in hearing improvements at 2000-8000Hz. The treatment effective rates were 44% and 80% for the non-postauricular injection group and postauricular injection group respectively, which showed a significant difference(χ²=8.385P<0.05).Conclusion:Postauricular injection of methylprednisolone as a remedy treatment is safe and effective for sudden hearing loss in middle and high frequency.

[Clinical analysis of hemorrhagic cystitis in children and adolescents with hematological diseases post haplo-hematopoietic stem cell transplantation].

Objective: To investigate the incidence and clinical features to probe the risk factors of hemorrhagic cystitis (HC) in children and adolescents with hematological diseases post haplo-hematopoietic stem cell transplantation (haplo-HSCT) . Methods: Medical records of 62 children and 27 adolescents with hematological diseases treated with haplo-HSCT between 2015 and 2016 were analyzed. Results: Of 89 cases (56 boys and 33 girls) , 44 patients were diagnosed with ALL, 33 AML, 3 AHL and 9 MDS. HC occurred in 32 of the 89 patients with an incidence of 36%, including 6 with grade Ⅰ, 16 with grade Ⅱ, 8 with grade Ⅲ, 2 with grade Ⅳ HC, respectively. The median time of HC onset was 25 days (range 2-55 days) after haplo-HSCT with the median duration as 19 days (range 3-95 days) , all of them were cured. The incidence of HC was lower in the group of children than that in the group of adolescents (27.4% vs 55.6%, χ(2)=6.466, P<0.05) , and the incidence of HC was higher in the group of patients who were ≥5 years old than that in the group of patients who were <5 years old (0 vs 34%, χ(2)=4.043, P<0.05) . Conclusion: HC is one of common complications in children and adolescents with hematological diseases post haplo-HSCT, older age was associated with increased mortality.

[The expression and clinical significance of RhoA, Ezrin and CD44 in laryngeal squamous cell carcinoma].

Objective:To investigate the expression of RhoA, Ezrin and CD44 proteins in laryngeal squamous cell carcinoma (LSCC), and analyze their roles and mutual relations in invasion and metastasis.Method:The expressions of RhoA, Ezrin and CD44 in 68 specimens of LSCC and 35 specimens of normal laryngeal mucosa tissues were detected by immunohistochemical SABC staining. The results were correlated with clinicopathological parameters.Result:The positive expression rates of RhoA, Ezrin and CD44 in LSCC were 80.9%,82,4% and 70.6%, and the positive expression rates of RhoA, Ezrin and CD44 in normal laryngeal mucosa tissues were 28.5%, 25.7% and 20%, respectively. The expressions of RhoA, Ezrin and CD44 were significantly correlated to depth of invasion (T), lymph node metastasis (N), clinical TNM stage, tumor grade respectively (P< 0.05), but not to age, gender, smoking, tumor location (P> 0.05). The expression of RhoA was positively correlated to that of Ezrin in LSCC, and the expression of Ezrin was positively correlated to that of CD44 in LSCC too (P< 0.05).Conclusion:Our findings suggested that high expressions of RhoA, Ezrin and CD44 might play an important role in the malignant progression of invasion and metastasis of LSCC, which indicated that they might have played synergistic effects in the process.

Statin-induced apoptosis of vascular endothelial cells is blocked by dexamethasone.

Statins block de novo synthesis of cholesterol by inhibiting the enzyme, HMG CoA reductase. The product of this reaction, mevalonic acid, is also a precursor of isoprenoids, molecules required for the activation of signalling G-proteins, such as Ras. Signal transduction pathways involving Ras are important for cell survival and this may be why statins induce apoptotic death of several cell types. Given that statins are used to treat vascular disease, it is surprising that no studies have been conducted on vascular endothelial cells. For this reason, we have tested the effect of fluvastatin (FS) on the endothelial cell line EA.hy 926. Here we show that FS, at concentrations from 1 to 2 microM, blocks growth and induces apoptosis of the endothelial cell line, EA.hy 926. As considerable redundancy exists in cell signalling pathways for cell survival, toxicity of FS under more physiological conditions might be prevented by pathways that do not require Ras, such as those activated by adrenal or sex steroids. To test this hypothesis, first RT-PCR analysis was performed for nuclear receptor mRNA expression. This revealed the presence of mRNA for the androgen receptor (AR) and glucocorticoid receptor (GR). The effect of the AR agonist, dihydrotestosterone (DHT), and the GR agonist, dexamethasone (Dex), was then tested. Whilst DHT (100 nM) had no effect on FS-induced cell death, Dex (1 microM) blocked FS-induced apoptosis. Cell cycle analysis revealed that 24 h exposure to FS prevented cells from leaving G(1) and 24-48 h later a marked sub-G(1) peak was observed. Dex was able to reduce the sub-G(1) peak, but it failed to reduce accumulation of cells in G(1). Further studies revealed that, in addition to blocking FS-induced apoptosis, Dex was able to block apoptosis of EA.hy 926 cells induced by serum deprivation, tumour necrosis factor-alpha, oxidants, DNA damage and mitochondrial disruption. This study strongly suggests that glucocorticoids have a role to play in preventing vascular injury and they may provide a reason why statins are apparently not toxic to vascular endothelial cells in vivo.

Selection of recombinant vaccinia viruses (Tian Tan strain) expressing hepatitis B virus surface antigen by using beta-galactosidase as a marker.

We constructed a plasmid that contains a small piece of DNA with two vaccinia promoters running in opposite directions--a promoter from a late gene encoding an 11 K polypeptide (P11) and a promoter from an early gene encoding 25K (P25). These promoters were isolated from the Tian Tan strain of vaccinia virus and were flanked by the thymidine kinase (TK) sequence of the same virus. Genes encoding the hepatitis B virus surface antigen (HBsAg) and the Escherichia coli beta-galactosidase (LacZ) were inserted downstream of the 11 K and 25 K promoters respectively so that coexpression plasmids were constructed. Recombinant vaccinia viruses were selected directly by picking blue plaques formed under overlaying agarose medium containing X-gal. HBsAg was expressed to high level by these recombinant viruses. These recombinant viruses showed reduced virulence on rabbit skin and induced anti-HBs after intradermal inoculation of rabbits.