The effects of statins in patients with advanced-stage cancers - a systematic review and meta-analysis.

Background: Statin therapy has been shown to reduce mortality in a wide range of cancer types and overall stages. Still, there is uncertainty about its efficacy in increasing survival among advanced cancer patients. Methods: We conducted a meta-analysis with data from all studies that compared the hazard ratio of overall survival, cancer-specific survival, and progression-free survival in patients with advanced-stage cancer who receive statin therapy. Studies were selected from the PubMed, Embase, and Web of Science databases from their inception to December 31, 2022. Cancer types are limited to those rarely screened during the annual examination and more likely to develop into advanced stages, such as lung, pancreatic and ovarian cancers. This resulted in 27 studies eligible for meta-analysis. Results: Statin therapy was associated with a 26% decreased risk of overall survival (HR, 0.74; 95% CI, 0.67, 0.81), 26% decreased risk of cancer-specific survival (HR, 0.74; 95% CI, 0.61-0.88), and 24% decreased risk of progression-free survival (HR, 0.76; 95% CI, 0.65-0.87) for advanced-stage cancer patients. The associations were not attenuated or reinforced by study design, study regions, cancer types, or other medical care. Concomitant use of other anticancer medications did not result in confounding effects. Conclusions: Statin therapy produces significant benefits on overall survival and cancer-specific survival. Although the benefits might be lower than the approved immunotherapy medications, its cost-effectiveness could lead to dramatic health consequences. Concomitant use of statin drugs as cancer treatments is highly recommended in future clinical trials.

Association between temperature variability and global meningitis incidence.

BACKGROUND: Meningitis can cause devastating epidemics and is susceptible to climate change. It is unclear how temperature variability, an indicator of climate change, is associated with meningitis incidence. METHODS: We used global meningitis incidence data along with meteorological and demographic data over 1990-2019 to identify the association between temperature variability and meningitis. We also employed future (2020-2100) climate data to predict meningitis incidence under different emission levels (SSPs: Shared Socioeconomic Pathways). RESULTS: We found that the mean temperature variability increased by almost 3 folds in the past 30 years. The largest changes occurred in Australasia, Tropical Latin America, and Central Sub-Saharan Africa. With a logarithmic unit increase in temperature variability, the overall global meningitis risk increases by 4.8 %. Australasia, Central Sub-Saharan Africa, and High-income North America are the most at-risk regions. Higher statistical differences were identified in males, children, and the elderly population. Compared to high-emission (SSP585) scenario, we predicted a median reduction of 85.8 % in meningitis incidence globally under the low-emission (SSP126) climate change scenario by 2100. CONCLUSION: Our study provides evidence for temperature variability being in association with meningitis incidence, which suggests that global actions are urgently needed to address climate change and to prevent meningitis occurrence.

E-Cigarette Usage and Arthritis in the United States, a Nationwide Cross-Sectional Survey.

Aim: The prevalence of the use of electronic cigarettes (e-cigarettes) has grown rapidly in the past decade in the United States. While numerous studies have demonstrated combustible cigarette is closely associated with an increased risk of arthritis diseases, little is known about the effect of e-cigarette usage on inflammatory arthritis diseases. We aimed to determinate if e-cigarette usage is associated with an increased risk of inflammatory arthritis. Methods: Data were obtained from the Behavioral Risk Factor Surveillance System, which is the largest national telephone-based survey of randomly sampled adults in the United States. A total of 924,882 participants with information on e-cigarette usage and inflammatory arthritis were included. We used multivariable logistic regression to estimate the risk of arthritis associated with e-cigarette usage. Results: Of the 924,882 participants, there were 30,569 (3.3%) current e-cigarette users, and 314,190 (25.9%) reported to have inflammatory arthritis diseases. In the fully adjusted model, we observed that the odds ratio (OR) (95% confidence interval) of inflammatory arthritis diseases was 1.81 (95% CI, 1.70-1.93) for current e-cigarette users compared with never e-cigarette users. The ORs of inflammatory arthritis diseases were 1.31 (95% CI, 1.18-1.47), and 1.55 (95% CI, 1.42-1.69) among sole e-cigarette and dual users compared with never e-cigarette users, respectively. Conclusions: This is the first study to observe a cross-sectional association between e-cigarette usage and inflammatory arthritis diseases, and the findings were consistent in both sole-e-cigarette users and dual users. Our findings provide evidence that e-cigarette usage might be an important risk factor for arthritis diseases, which may have regulatory implications for e-cigarette control.

The Association Between E-Cigarette Use and Prediabetes: Results From the Behavioral Risk Factor Surveillance System, 2016-2018.

INTRODUCTION: Both E-cigarette use and the prevalence of prediabetes have risen dramatically in the past decade. It is crucial to understand whether E-cigarette use is associated with the risk of prediabetes. METHODS: Participants who completed the prediabetes and E-cigarette modules of the Behavioral Risk Factor Surveillance System survey (2016-2018) were included in this study. E-cigarette use information was collected by asking: Have you ever used an e-cigarette or other electronic "vaping" product, even just one time, in your entire life? We defined sole E-cigarette users as current E-cigarette users who are never combustible-cigarette users, and dual users were defined as both current E-cigarette and combustible-cigarette users. Participants with prediabetes were identified by asking: Ever been told by a doctor or other health professional that you have prediabetes or borderline diabetes? Multivariable logistic regression was used to determine the association between E-cigarette use and prediabetes. RESULTS: Among the 600,046 respondents, 28.6% of respondents were aged <35 years. The prevalence of prediabetes among current E-cigarette, sole E-cigarette users, and dual users was 9.0% (95% CI=8.6, 9.4), 5.9% (95% CI=5.3, 6.5), and 10.2% (95% CI=9.8, 10.7), respectively. In the fully adjusted model, the ORs for prediabetes were 1.22 (95% CI=1.10, 1.37) for current E-cigarette users and 1.12 (95% CI=1.05, 1.19) for former E-cigarette users compared with that of never E-cigarette users. The ORs for prediabetes were 1.54 (95% CI=1.17, 2.04) for sole E-cigarette users and 1.14 (95% CI=0.97, 1.34) for dual users. CONCLUSIONS: In this representative sample of U.S. adults, E-cigarette use was associated with greater odds of prediabetes. The results were consistent in sole E-cigarette users.

Phenolic Acids-Rich Fractions from Agaricus bitorguis (Quél.) Sacc. Chaidam ZJU-CDMA-12 Mycelia Modulate Hypoxic Stress on Hypoxia-Damaged PC12 Cells.

Hypoxia is a common pathological process in various clinical diseases. However, there is still a lack of effective anti-hypoxia active substances. Agaricus bitorguis (Quél.) Sacc Chaidam (ABSC) is a rare wild edible macrofungus that grows underground at high altitudes. Herein, intracellular phenolic acids-rich fractions (IPA) were extracted from ABSC ZJU-CDMA-12, and the structural characterization and anti-hypoxia activity of IPA on PC12 cells were elucidated as well. The results of HPLC-Q-TOF-MS illustrated that five kinds of IPA were isolated from ABSC, including (-)-epicatechin gallate, arabelline, yunnaneic acid D, 2'-O-p-hydroxybenzoyl-6'-O-trans-caffeoylgardoside,4'-O-methylgallocatechin-(4->8)-4'-O-methylepigallocatechin. IPA extracted from ABSC proved to show anti-hypoxia activity on hypoxia-damaged PC12 cells. Hypoxia enhanced reactive oxygen species (ROS) generation and reduced the mitochondrial membrane potential (ΔΨm) in PC12 cells, resulting in the inhibition of survival and induction of apoptosis in PC12 cells. Measurements of 100 µg/mL and 250 µg/mL IPA could significantly reduce hypoxia-induced damage in PC12 cells by decreasing overproduced intracellular ROS, improving ΔΨm, and reducing cell apoptosis rate. Our findings indicated that the IPA from ABSC potentially could be used as novel bioactive components applied to anti-hypoxia functional foods or medicines.

Chinese Yellow Rice Wine Processing with Reduced Ethyl Carbamate Formation by Deleting Transcriptional Regulator Dal80p in Saccharomyces cerevisiae.

Ethyl carbamate (EC) is a potential carcinogen that forms spontaneously during Chinese rice wine fermentation. The primary precursor for EC formation is urea, which originates from both external sources and arginine degradation. Urea degradation is suppressed by nitrogen catabolite repression (NCR) in Saccharomyces cerevisiae. The regulation of NCR is mediated by two positive regulators (Gln3p, Gat1p/Nil1p) and two negative regulators (Dal80p/Uga43p, Deh1p/Nil2p/GZF3p). DAL80 revealed higher transcriptional level when yeast cells were cultivated under nitrogen-limited conditions. In this study, when DAL80-deleted yeast cells were compared to wild-type BY4741 cells, less urea was accumulated, and genes involved in urea utilization were up-regulated. Furthermore, Chinese rice wine fermentation was conducted using dal80Δ cells; the concentrations of urea and EC were both reduced when compared to the BY4741 and traditional fermentation starter. The findings of this work indicated Dal80p is involved in EC formation possibly through regulating urea metabolism and may be used as the potential target for EC reduction.

Effects of acupuncture treatment on motor function in patients with subacute hemorrhagic stroke: A randomized controlled study.

OBJECTIVE: The aim of this study was to evaluate the additional effects of acupuncture treatment on motor function in patients with subacute hemorrhagic stroke. DESIGN: One hundred and thirty-four patients with subacute hemorrhagic stroke were randomized to receive acupuncture treatment plus conventional treatment (treatment group) or conventional treatment only (control group). Acupuncture treatments were given in 24 sessions over 4 weeks, with 3-month follow-up period. Blinded evaluation was based on Fugl-Meyer Assessment, Barthel Index with an intention-to-treat analysis. For those patients who were able to walk, a three-dimensional gait analysis system was employed to objectively record spatiotemporal and kinematic parameters. RESULTS: Compared with control group, the treatment group showed a significantly greater over-time improvement in total Fugl-Meyer, lower-limb Fugl-Meyer, but not in upper-limb Fugl-Meyer and Barthel Index. The spatiotemporal parameters of velocity, step length, cadence, step width all showed significant difference between the 2 groups. The velocity in treatment group decreased unexpectedly at day 14, then increased sharply and overcame control group at day 28. The treatment group also showed a significantly greater increase in peak circumduction, peak hip hiking, hip range of motion, knee range of motion and a tendency for the ankle range of motion. CONCLUSIONS: Acupuncture may promote the motor function recovery of hemorrhagic stroke patients in subacute phase mainly by enhancing the lower limb ability. It probably diminishes the compensation strategies earlier to correct the abnormal gait pattern. Although this adjustment may result in a compromise in the improvement of gait velocity temporarily, patients would benefit from it in a long run.

Statin-induced GGPP depletion blocks macropinocytosis and starves cells with oncogenic defects.

Cancer cells display novel characteristics which can be exploited for therapeutic advantage. Isolated studies have shown that 1) the mevalonate pathway and 2) increased macropinocytosis are important in tumorigenesis, but a connection between these two observations has not been envisioned. A library screen for compounds that selectively killed Dictyostelium pten- cells identified pitavastatin. Pitavastatin also killed human breast epithelial MCF10A cells lacking PTEN or expressing K-RasG12V, as well as mouse tumor organoids. The selective killing of cells with oncogenic defects was traced to GGPP (geranylgeranyl diphosphate) depletion. Disruption of GGPP synthase in Dictyostelium revealed that GGPP is needed for pseudopod extension and macropinocytosis. Fluid-phase uptake through macropinocytosis is lower in PTEN-deleted cells and, as reported previously, higher in cells expressing activated Ras. Nevertheless, uptake was more sensitive to pitavastatin in cells with either of these oncogenic mutations than in wild-type cells. Loading the residual macropinosomes after pitavastatin with high concentrations of protein mitigated the cell death, indicating that defective macropinocytosis leads to amino acid starvation. Our studies suggest that the dependence of cancer cells on the mevalonate pathway is due to the role of GGPP in macropinocytosis and the reliance of these cells on macropinocytosis for nutrient uptake. Thus, inhibition of the networks mediating these processes is likely to be effective in cancer intervention.

Lidocaine inhibits melanoma cell proliferation by regulating ERK phosphorylation.

The melanoma is responsible for the majority of all skin cancer-related deaths worldwide. Evidence suggests that local anesthetics provide some benefit in the treatment of cancer via inhibition of cellular proliferation, invasion and migration. However, the potential antiproliferative effects of local anesthetics in the treatment of melanoma remain to be elucidated. In this study, we investigated the antiproliferative effects and underlying mechanism of the commonly used local anesthetic (lidocaine) on melanoma cells. A375 melanoma cells were treated by lidocaine or vemurafenib. Cell Counting Kit-8, histological staining, flow cytometric analysis, immunohistochemical staining, and Western blot analyses were carried out to test the effects of lidocaine and vemurafenib on A375 cells. BALB/C-nu/nu mice intraperitoneally injected with A375 cells were treated by lidocaine, and then tumor volume and weight were calculated. Lidocaine exhibited vemurafenib-like effects totally. Lidocaine inhibited A375 melanoma cell proliferation in a dose- and time-dependent manner and colony formation also showed a dose-dependent inhibition. Lidocaine treatment resulted in the arrest of cell-cycle progression in the G1 phase and inhibited Ki-67 expression in a dose-dependent manner. This effect was associated with inhibited extracellular signal-regulated kinase (ERK) phosphorylation. In vivo experiments revealed that intravenous injections of lidocaine suppressed tumor volume and weight. Lidocaine inhibits melanoma cell proliferation in a dose- and time-dependent manner via a mechanism that may involve inhibition of the ERK signaling pathway. Thus, lidocaine may provide some benefit for the treatment of melanoma.

Immediate effect of scalp acupuncture on the gait of patients with subacute intracerebral haemorrhage analysed by three-dimensional motion: secondary analysis of a randomised controlled trial.

OBJECTIVE: To investigate the immediate effect of scalp acupuncture on walking pattern, using three-dimensional gait analysis (3D-GA), among patients in the subacute stage of intracerebral haemorrhage (ICH). METHODS: A subset of 30 patients with subacute ICH participating in a recently published randomised controlled trial who were able to walk independently were assessed by 3D-GA before and immediately after scalp acupuncture treatment (treatment group) or no intervention (control group) and the results presented here as a secondary analysis. The acupuncture manipulation was repeated three times with an interval of 5 min. Spatiotemporal and kinematic parameters during walking were collected and analysed using a 3D motion analysis system. RESULTS: After treatment, there were significant differences between the treatment and control groups in the spatiotemporal parameters of step length, velocity and cadence (p<0.05) and double-limb support. No significant difference was found in step width. When kinematic parameters were evaluated, the treatment group showed a significantly decreased peak pelvic anterior tilt angle and an increased hip extension angle after scalp acupuncture treatment, whereas the control group demonstrated no temporal changes. There were no significant changes in any other kinematic parameters in either group. CONCLUSIONS: As the first exploratory study to investigate the effect of the scalp acupuncture on gait performance in patients with subacute ICH, this secondary analysis of a recent randomised trial suggested an immediate effect of treatment on spatiotemporal parameters. Improvement in gait pattern may be associated with a decreased anterior tilt of the pelvis and augmented hip joint motion during walking. TRIAL REGISTRATION NUMBER: ChiCTR-TRC-08000225; Post-results.

Anti-apoptosis endothelial cell-secreted microRNA-195-5p promotes pulmonary arterial smooth muscle cell proliferation and migration in pulmonary arterial hypertension.

In the pathological mechanism of pulmonary arterial hypertension, the role of apoptosis-resistant pulmonary microvascular endothelial cells (PVECs/AR) has been emphasized on the pulmonary vascular remodeling. In the present study, we investigated whether PVECs/AR can promote the proliferation and migration of pulmonary arterial smooth muscle cells (PASMCs), and to study the role of miR-195-5p in the crosstalk between these two types of cells. We confirmed that PVECs/AR can promote the proliferation and migration of PASMCs in a co-culture system of AR/PVECs and PASMCs. Additionally, after exposure to hypoxia for 12 or 24 h, AR/PVECs had a higher mature miR-195-5p level than PVECs (P < 0.05, 12 and 24 h). Luciferase reporter assays were used to validate indications of the existence of an HRE in the miR-195-5p promoter. Knocking down Smad7 can reverse the inhibition of Lv-S195 on TGF-ß1-induced PASMCs remodeling. TGF-ß1 promoted cell growth in PASMCs, and the supernatant of PVECs/AP infected with Lv-S195 inhibited TGF-ß1 enhanced proliferation in PASMCs, which was also blocked by Lv-shRNA-Smad7. The result of this experiment confirmed the specificity of the HIF-1a/miR-195/Smad7 pathway. Our data indicate the possible function of PVECs/AR in the process of pulmonary vascular remodeling. MiRNA-195-5p played a role as an interacting paracrine factor between PVECs/AR and PASMC, which seemed to function through the HIF-1a/miRNA-195-5p/Smad7 pathway.

Efficacy and safety of penetration acupuncture on head for acute intracerebral hemorrhage: A randomized controlled study.

BACKGROUND: Acupuncture, especially acupuncture treatment on head for acute intracerebral hemorrhage (ICH), has long been disputable. The aim of this study was to evaluate the efficacy and safety of penetration acupuncture on head in patients with acute ICH. METHODS: Eighty-two patients with acute ICH were randomized to receive penetration acupuncture treatment on head combined with conventional treatment (treatment group [TG]) or conventional treatment only (control group [CG]). Acupuncture treatments were given in 24 sessions over 4 weeks, with 3-month follow-up period. Measures included Clinical Neurological Function Deficit Scale (CNFDS), Barthel Index (BI), vital signs (respiration, heart rate, blood pressure, and oxygen saturation), and hematoma absorption ratio. RESULTS: Both groups showed a progressively improvement in CNFDS and BI scores from day 7 to 90. The TG showed a significantly greater improvement in CNFDS than CG over time (P < 0.05). However, BI failed to show significant difference between the 2 groups (P > 0.05). The vital signs were stable and no expansion of hematoma occurred over the course of acupuncture treatment. CONCLUSION: Penetration acupuncture treatment on head appeared to be safe over the course of treatment on acute ICH and may result in additional functional improvements detected in the CNFDS but not reflected in the BI. A larger-scale clinical trial with longer follow-up assessments is required to confirm these findings.

[Immediate effects of twirling reinforcing-reducing manipulation at head acupoints on muscle force in patients with acute ischemic stroke: a randomized controlled trial].

OBJECTIVE: To observe differences of immediate effect of twirling reinforcing-reducing manipulation at head acupoints on muscle force in patients with acute ischemic stroke. METHODS: A total of 126 patients who met the inclusive criteria were randomly divided into a twirling reinforcing group, a mild reinforcing-reducing group and a twirling reducing group, 42 cases in each one. The lines between Baihui (GV 20) and bilateral Taiyang (EX-HN 5) were selected and treated with successive insertion of three needles. The twirling reinforcing method was used in the twirling reinforcing group, and mild reinforcing-reducing method was used in the mild reinforcing- reducing group, and twirling reducing method was used in the twirling reducing group. Each needle was manipulated for 1 min, which was repeated once every 10 min. The needles were retained for 30 min. Muscle force was evaluated immediately after treatment. RESULTS: (1) Muscle force of upper limb: after acupuncture, the muscle force of proximal and remote ends were all improved significantly in three groups (all P < 0.05), which was more obvious in proximal end (all P < 0.05). The improvement of the muscle force of proximal end in the twirling reinforcing group was superior to those in the mild reinforcing-reducing group and twirling reducing group (both P < 0.05); the differences of the muscle force of remote end were not significant among three groups (all P > 0.05). (2) Muscle force of lower limb: after acupuncture, the muscle force of proximal and remote ends was all improved significantly in the three groups (all P < 0.05). The improvement of the muscle force of proximal end was superior to that of remote end in the twirling reinforcing group and mild reinforcing-reducing group (both P < 0.05). CONCLUSION: The twirling reinforcing manipulation at head acupoints is superior to mild reinforcing-reducing and twirling reducing manipulation for muscle force in patients with acute ischemic stroke, which is more significant in proximal end of limbs.

[Impacts on fine movement of upper extremity with flaccid paralysis in stroke treated with acupuncture combined with motor imagery].

OBJECTIVE: To explore the impacts on the fine movement of upper extremity with flaccid paralysis in stroke treated with acupuncture and motor imagery. METHODS: Sixty-two cases of flaccid paralysis of upper extremity in stroke were randomized into an observation group (30 cases) and a control group (32 cases). In the control group, the conventional western medication and the passive movement of the extremity were applied. Additionally, the penetrating needling technique was used at the head points [penetrating needling from Baihui (GV 20) to Taiyang (EX-HN 5)] and the local affected extremity [penetrating needling from Jianyu (LI 15) to Binao (LI1 14), from Quchi (LI 11) to Shaohai (HT 3), from Waiguan (TE 5) to Neiguan (PC 6), etc]. The needles were retained for 30 min. In the observation group, on the basic treatment as the control group, during the needle retaining, the motor imagery therapy was supplemented. The treatment was given once a day, 5 treatments a week, for 4 weeks totally in the either group. The scores in the action research arm test (ARAT) and the modified Fugl-Meyer assessment (FMA) were recorded at the moment of enrollment, in 2 and 4 weeks of treatment, in the 6th and 8th weeks, as well as after 90 days separately. RESULTS: (1) ARAT score: since the 2nd week, ARAT scores in the observation group were all improved significantly as compared with those in the control group at the each time points (all P<0. 05). In the observation group, the scores were improved significantly in the pair comparison at the time points (all P<0. 05). In the control group, the score in the 2nd week was not different significantly as compared with that before treatment (P>0. 05), but the scores at the rest time points were improved significantly in the pair comparison (all P<0. 05); (2) FMA score: in the 2nd week and on the 90th day, FMA score in the observation group was higher significantly than those in the control group (both P < 0. 05). In the observation group, the scores were all improved significantly in the pair comparison at the time points (all P<0. 05). In the control group, except that in comparison between the 90th day and the 8th week (P>0. 05), the results were all P<0. 05 at the rest time points. CONCLUSION: The early intervention of the combined therapy of acupuncture and motor imagery effectively promotes the recovery of the coordination function and the fine movement of upper extremity, especially the improvements of the fine movement in stroke at flaccid paralysis stage. The efficacy is better than the single acupuncture treatment.

In Vitro Antioxidant Activities and in Vivo Anti-Hypoxic Activity of the Edible Mushroom Agaricus bisporus (Lange) Sing. Chaidam.

With the rising awareness of a healthy lifestyle, natural functional foods have gained much interest as promising alternatives to synthetic functional drugs. Recently, wild Agaricus bisporus (Lange) Sing. Chaidam has been found and artificially cultivated for its thick fresh body and excellent taste, with its antioxidant and anti-hypoxic abilities unknown. In this work, the antioxidant potential of its methanolic, 55% ethanolic, aqueous extracts and crude polysaccharide was evaluated in different systems. The results showed that polysaccharide was the most effective in scavenging ability on 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl radicals, metal chelating activity and reducing power, with EC50 values of 0.02, 2.79, 1.29, and 1.82 mg/mL, respectively. Therefore, we further studied the anti-hypoxic activity of crude polysaccharide. The results turned out that polysaccharide (300 mg/kg) prolonged the survival time, decreased the blood urea nitrogen and lactic acid content as well as increased the liver glycogen significantly, compared with the blank control and the commercialized product Hongjingtian (p < 0.05). With such excellent activities, we purified the polysaccharide and analyzed its molecular weight (120 kDa) as well as monosaccharide components (glucose, fructose and mannose). This study indicated that wild Agaricus bisporus (Lange) Sing. Chaidam had strong potential to be exploited as an effective natural functional food to relieve oxidative and hypoxia stresses.

Optimization of culture medium compositions for gellan gum production by a halobacterium Sphingomonas paucimobilis.

The effect of culture medium compositions on gellan gum production produced by fermentation with a halobacterium Sphingomonas paucimobilis QHZJUJW CGMCC2428 was studied. In this work, a fractional factorial design was applied to investigate the main factors that affected gellan gum production by S. paucimobilis QHZJUJW CGMCC2428. Sucrose was the best carbon source for gellan gum and peptone displayed better inducing effect. Central composite design and response surface methodology were adopted to derive a statistical model for optimizing submerged culture medium composition. These experimental results showed that the optimum culture medium for producing gellan gum was composed of 40.00 (w/v) sucrose, 3.00% peptone (w/v), MgSO4 (w/v), 9.20% KH2PO4 (w/v), 7.50% Na2HPO4 (w/v), 4.30% K2SO4 (w/v), pH 6.8-7.0. The maximal gellan gum was 19.89±0.68 g/L, which was agreed closely with the predicated value (20.12 g/L). After incubated for 72 h under the optimized culture medium in 5-L bioreactor, the gellan gum fermentation reached about 19.90±0.68 g/L, which was higher than that in the initial cultivation medium.

The cross talk between cGMP signal pathway and PKC in pulmonary endothelial cell angiogenesis.

Angiogenic proliferation of vascular endothelial cells is believed to play an important role in pulmonary vascular remodeling in pulmonary arterial hypertension. In the present study, we found that c-GMP (cyclic guanosine monophosphate) inhibited the proliferation and tube formation of pulmonary vascular endothelial cells induced by TGF-ß1, and that this process was reversed by PKG (protein kinase G) inhibitor and PKC (protein kinase C) inhibitor. In addition, small interfering RNA (siRNA) targeting ERK also reduced cellular proliferation. Furthermore, western blotting showed that cGMP down-regulated the phosphorylation level of ERK1/2, which was reversed not only by PKG inhibitor but also by PKC inhibitor. Silencing different PKC isoforms showed that PKCΔ, PKCγ and PKCα were involved in ERK phosphorylation, suggesting that PKC kinases have a permissive action. Three subtypes, PKCΔ, PKCγ and PKCα are likely to be involved the phosphorylation suppression of ERK included cGMP. Taken together, these data suggest that ERK phosphorylation mediates the proliferation of pulmonary vascular endothelial cells, and PKC kinases have a permissive action in this process.

Ethyl Carbamate in Fermented Beverages: Presence, Analytical Chemistry, Formation Mechanism, and Mitigation Proposals.

Ethyl carbamate (EC) commonly found in fermented beverages has been verified to be a multisite carcinogen in experimental animals. EC was upgraded to Group 2A by the Intl. Agency for Research on Cancer (IARC) in 2007, which indicates that EC is a probable carcinogen to humans. Because of its threat to human safety, the presence of EC may be a big challenge in the alcoholic beverage industry. During the past few years, thorough and systematic research has been carried out in terms of the generation of EC in order to meet the allowed limitation levels in fermented beverages. Previous studies have indicated that EC primarily results from the reaction of ethanol and compounds containing carbamyl groups. These main EC precursors are commonly generated from arginine metabolism by Saccharomyces cerevisiae or lactic acid bacteria accompanied by the fermentation process. This review comprehensively summarizes the genotoxicity, analytical methods, formation pathways, and removal strategies of EC in various beverages. The article also presents the metabolic mechanism of EC precursors and pertinent metabolites, such as urea, citrulline, and arginine.

A versatile mini-mazF-cassette for marker-free targeted genetic modification in Bacillus subtilis.

There are some drawbacks for MazF-cassette constructed in previous reports for marker-free genetic manipulation in Bacillus subtilis, including cloning-dependent methodology and non-strictly controlled expression system. In our study, the modifications on mazF-cassette are carried out, such as using mini Zeocin resistance gene as positive-selectable marker and strictly controlled xyl promoter from the B. subtilis to replace non-strictly controlled IPTG-inducible Pspac or xyl promoter from Bacillus megaterium. Then the mini-mazF-cassette was successfully applied to knock-out the amyE gene, to delete a 90-kb gene cluster, and to knock-in a green fluorescent protein expression cassette employing a cloning-independent methodology, without introducing undesirable redundant sequences at the modified locus in the B. subtilis 1A751. Besides, the mini-mazF-cassette could be used repeatedly to delete multiple genes or gene clusters with only a 2- to 2.5-kb PCR-fused fragment, which largely reduced the frequency of nucleic acid mutations generated by PCR compared to previous reports. We further demonstrated that the frequency of spontaneous mazF-resistant mutants was lower, and the frequency of generating desired clones was nearly 100%. The entire procedure for marker-free genetic manipulation using the mini-mazF-cassette can be finished in about 3days. This modified cassette has remarkable improvement compared to existing approaches and is applicable for available manipulating Bacillus species chromosomes.