Optimization of lactic acid bacterial starter culture to improve the quality and flavor characteristics of traditional Hurood.

Hurood is a traditional fermented milk product prepared by traditional Mongolian techniques of fermenting raw milk, partial degreasing, heating, whey drainage, emulsification of curd, and molding. Currently, Hurood available in the market is generally prepared by small-scale enterprises at home or in open air. Therefore, lack of standardization of bacterial starter culture leads to variation in the flavor and sensory properties of Hurood from batch to batch. In this study, we aimed to assess the best starter culture combination obtained from 37 lactic acid bacterial strains isolated from traditional Hurood. The solidification state and sensory quality were used as indexes for determining the fermentation efficiency of the bacterial starter culture combinations. The yield and texture characteristics were used to determine the optimal ratio of bacterial strains in a combination and the processing conditions for traditional Hurood production. The most optimal bacterial culture combination was observed to be NF 9-3:NF 10-4:CH 3-1 in 5:4:1 ratio and in 3% amount. The most optimal whey temperature and heating-stirring temperature were observed to be 55°C to 60°C and 85°C to 90°C, respectively. Hurood prepared with the optimal combination of bacterial strains exhibited significantly enhanced sensory quality, flavor, and contents of AA and fatty acids. Therefore, the use of optimal starter culture of lactic acid bacteria could produce Hurood with significantly superior sensory qualities, making the product more acceptable to consumers.

RNA splicing analysis contributes to reclassifying variants of uncertain significance and improves the diagnosis of monogenic disorders.

BACKGROUND: Numerous variants of uncertain significance (VUSs) have been identified by whole exome sequencing in clinical practice. However, VUSs are not currently considered medically actionable. OBJECTIVE: To assess the splicing patterns of 49 VUSs in 48 families identified clinically to improve genetic counselling and family planning. METHODS: Forty-nine participants with 49 VUSs were recruited from the Reproductive and Genetic Hospital of CITIC-Xiangya. Bioinformatic analysis was performed to preliminarily predict the splicing effects of these VUSs. RT-PCR and minigene analysis were used to assess the splicing patterns of the VUSs. According to the results obtained, couples opted for different methods of reproductive interventions to conceive a child, including prenatal diagnosis and preimplantation genetic testing (PGT). RESULTS: Eleven variants were found to alter pre-mRNA splicing and one variant caused nonsense-mediated mRNA decay, which resulted in the reclassification of these VUSs as likely pathogenic. One couple chose to undergo in vitro fertilisation with PGT treatment; a healthy embryo was transferred and the pregnancy is ongoing. Three couples opted for natural pregnancy with prenatal diagnosis. One couple terminated the pregnancy because the fetus was affected by short-rib thoracic dysplasia and harboured the related variant. The infants of the other two couples were born and were healthy at their last recorded follow-up. CONCLUSION: RNA splicing analysis is an important method to assess the impact of sequence variants on splicing in clinical practice and can contribute to the reclassification of a significant proportion of VUSs. RNA splicing analysis should be considered for genetic disease diagnostics.

Rapid Drug-Screening Platform Using Field-Effect Transistor-Based Biosensors: A Study of Extracellular Drug Effects on Transmembrane Potentials.

Ion channel-modulating drugs play an important role in treating cardiovascular diseases. Facing the demands for continuous monitoring of drug effectiveness, the conventional techniques have become limited when investigating a long-term cellular physiology. To address the challenge, we propose a drug-screening platform using the stretch-out electrical double layer (EDL)-gated field-effect transistor-based biosensors (BioFETs). In this work, BioFETs were utilized to amplify electrophysiological signals from the mammalian cardiomyocytes (H9c2). The stretch-out configuration avoided a chemical corrosion on FETs and prolonged the lifetime of a BioFET system. A physical model is presented to elucidate the signal response to a drug effect on a cell. Fibronectin and gelatin were coated on sensors and served as the adhesive layers where H9c2 cells attached. BioFETs demonstrated an ability to qualitatively distinguish a depolarization and a polarization of the cytomembranes. The signal responses to the changes of transmembrane potentials were monitored in real-time, and they were highly correlated. The effects of nifedipine and calcium ions on cellular electrophysiology were examined and discussed. Due to the capability of a rapid detection, a prolonged lifetime, and an excellent sensitivity to an electrical change, a stretch-out EDL-gated BioFET can be a drug-screening platform for ion channel modulators.

High-Sensitivity and Trace-Amount Specimen Electrochemical Sensors for Exploring the Levels of ß-Amyloid in Human Blood and Tears.

As the occurrence of Alzheimer's disease (AD) has increased, the detection and treatment of AD have become global social issues. Effective early detection and wide-range screening of AD allow patients to gain early control and delay brain degeneration. For these reasons, we choose electrochemical sensors to complete the detection task. Although bio-electrochemical technology for antibody and antigen sensing is not a new technology, considering the scarcity of tear samples for dementia and since the existing AD detection techniques are highly invasive and expensive for subjects, we have to use the traditional detection techniques for the early screening of Alzheimer's disease via trace-amount specimens. An AD-related protein in the eye is thought to be an important biomarker for early detection. To carry out detection using tear samples as a test specimens, a tear collection device was developed in this study that extracted 10 µL of tear fluid from a tear Schirmer strip. In this research, we distinguished healthy people in different age groups and detect Aß in both tear and blood samples. We developed a biosensor, which could detect Aß in tear specimen from 1 to 100 pg/mL. Also, this biosensor is inexpensive, disposable, and easy to use. In our result, the concentration of Aß in tears was approximately 10 times more than that in blood. This study demonstrates the feasibility and prospects of future screening for AD-associated biomarkers by a dynamic comparison between blood and tears.

Protective effects of four new saponins from Gynostemma pentaphyllum against hydrogen peroxide-induced neurotoxicity in SH-SY5Y cells.

Gynostemma pentaphyllum has been used as a medicine-food homologious health product in China for a long time. This research aimed to isolate and identify its active compounds with protective effects against hydrogen peroxide induced SH-SY5Y cell death. Four new dammarane-type saponins were isolated from G. pentaphyllum using various chromatographic methods. They were identified as gypenoside S1 (1), gypenoside S3 (2), gypenoside S2 (3) and gypenoside S4 (4), respectively by HRESIMS and NMR spectra. Their cytotoxic activity was evaluated against three human cancer cell lines, A549 (lung), HepG2 (liver), SH-SY5Y (nerve), by MTT method. They showed low cytotoxicities with the IC50 values of more than 100 µM on three cancer cell lines. However, they appeared protective effects against hydrogen peroxide induced SH-SY5Y cell death in a dose-dependent manner. They recovered cell viability more than 69% at the concentration of 20 µM from 66%, while as vitamin C to 67%. Compound 3 and 4 recovered more than 79% at 100 µM. The present study suggests that G. pentaphyllum has antioxidative potential and the saponins from G. pentaphyllum are considered as the active compounds with safe and neuroprotecitve effect.

[Protective effects of flavonoids from Gynostemma pentaphyllum on oxidative damage in LLC-PK1 cells].

Four flavonoids were isolated from Gynostemma pentaphyllum by chromatography methods and their structures were identified by MS and NMR spectra data as quercetin-3-O-( 2″,6″-di-α-L-rhamnosyl)-ß-D-galactopyranoside( 1),quercetin-3-O-( 2″,6″-di-α-L-rhamnosyl)-ß-D-glucopyranoside( 2),quercetin-3-O-( 2″-α-L-rhamnosyl)-ß-D-galactopyranoside( 3),and quercetin-3-O-( 2″-α-L-rhamnosyl)-ß-D-glucopyranoside( 4). Among them,compounds 1-3 were obtained from the Cucurbitaceae family for the first time.The four flavonoids showed potent antioxidant effects against the DPPH,·OH and ■radicals in vitro,especially for DPPH radical scavenging activity with the IC50 values of 71. 4,29. 5,48. 3 and 79. 2 µmol·L~(-1),respectively. Moreover,the four flavonoids displayed strong cytoprotection against AAPH-induced oxidative damage in LLC-PK1 cells by suppressing the increase of malondialdehyde( MDA) and the decrease of the superoxide dismutase( SOD) and glutathione( GSH). Since further research is needed to prove its efficacy in vivo and clinical trial,the study may provide four potential antioxidants from G. pentaphyllum.

[Effects of flavonoids from Gynostemma pentaphyllum on A549 cells damaged by hydrogen peroxide].

This study focuses on the therapeutical effect of flavonoids from Gynostemma pentaphyllum on human lung carcinoma A549 cells induced by H2O2 oxidative stress and its possible mechanisms. The oxidative damage model was established using different concentrations H2O2 to induce A549 cell for different hours, and then treated with the flavonoids for 10 hours. The effects of flavonoids from G. pentaphyllum on cell viability of A549 cell damaged by H2O2 were detected by MTT assay. The contents of ROS were detected by DCFH-DA fluorescent probe method via flow cytometer. The contents of MDA, SOD and GSH were detected by TBA，NBT and DTNB-linked colorimetry assay, respectively. Expressions levels of Nrf2, NQO1 and HO-1 in A549 cells were evaluated by Western blot. The results showed that the cell activity was decreasing with the rise of H2O2 concentration within the range of 200-700 µmol·L⁻¹. The cell viability was 60.4% after treated with 500 µmol·L⁻¹H2O2 for 10 h, so it was chosen to be as an oxidant stress model. Compared with normal group，the contents of SOD, GSH and HO-1 expressions were lower after damaged with H2O2. On the contrary, the contents of ROS and MDA expressions were increased. Compared with model group, the contents of SOD, GSH and the expressions of Nrf2, NQO1 and HO-1 were increased after treated with flavonoids from G. pentaphyllum. The above results demonstrate that flavonoids from G. pentaphyllum may attenuate the effect of H2O2-induced oxidative stress on A549 cell by resisting oxidation. The finding may provide a biological evidence for the application of the G. pentaphyllum to fight the oxidative stress related diseases.

A new dammarane-type saponin from Gynostemma pentaphyllum induces apoptosis in A549 human lung carcinoma cells.

Gynostemma pentaphyllum has been widely used as a traditional herb for its antioxidant and immunostimulatory activities. We have previously reported several useful dammarane-type saponins with cytotoxicity against A549 human lung cancer cells from heat-processed G. pentaphyllum. In this study, a new dammarane-type saponin, 20(S)-2α,3ß,12ß-tetrahydroxydammar-3-O-ß-d-glucopyranoside (namely gypenoside Jh1), was isolated from the ethanol extract of heat-processed G. pentaphyllum using column chromatography and semi-preparative HPLC. Gypenoside Jh1 exhibited strong cytotoxicity against A549 cells in a concentration-dependent manner, which was associated with apoptotic cell death characterized by morphological changes, Hoechst 33258 nuclear staining, Annexin V and propidium iodide binding and mitochondrial potentials assay. Quantitative analysis using flow cytometry also showed that the proportion of apoptotic cells was increased after gypenoside Jh1 treatment. These findings indicated that gypenoside Jh1 showed antiproliferative effects on A549 cells and mitochondrial-dependent pathway is involved in gypenoside Jh1-induced apoptosis.

Deoxypodophyllotoxin induces G2/M cell cycle arrest and apoptosis in SGC-7901 cells and inhibits tumor growth in vivo.

Deoxypodophyllotoxin (DPT), a natural microtubule destabilizer, was isolated from Anthriscus sylvestris, and a few studies have reported its anti-cancer effect. However, the in vivo antitumor efficacy of DPT is currently indeterminate. In this study, we investigated the anti-gastric cancer effects of DPT both in vitro and in vivo. Our data showed that DPT inhibited cancer cell proliferation and induced G2/M cell cycle arrest accompanied by an increase in apoptotic cell death in SGC-7901 cancer cells. In addition, DPT caused cyclin B1, Cdc2 and Cdc25C to accumulate, decreased the expression of Bcl-2 and activated caspase-3 and PARP, suggesting that caspase-mediated pathways were involved in DPT-induced apoptosis. Animal studies revealed that DPT significantly inhibited tumor growth and decreased microvessel density (MVD) in a xenograft model of gastric cancer. Taken together, our findings provide a framework for further exploration of DPT as a novel chemotherapeutic for human gastric cancer.

Formulations, hemolytic and pharmacokinetic studies on saikosaponin a and saikosaponin d compound liposomes.

The aim of this study was to develop and optimise a saikosaponin a and saikosaponin d compound liposome (SSa-SSd-Lip) formulation with reduced hemolysis and enhanced bioavailability. A screening experiment was done with Plackett-Burman design, and response surface methodology of five factors (EPC/SSa-SSd ratio, EPC/Chol ratio, water temperature, pH of PBS, and ultrasound time) was employed to optimise the mean diameter, entrapment efficiency of SSa and SSd, and the reduction of hemolysis for SSa-SSd-Lip. Under the optimal process conditions (EPC/SSa-SSd ratio, EPC/Chol ratio, water temperature and pH of PBS were 26.71, 4, 50 °C and 7.4, respectively), the mean diameter, the entrapment efficiency of SSa, the entrapment efficiency of SSd and the hemolysis were 203 nm, 79.87%, 86.19%, 25.16% (SSa/SSd 12.5 mg/mL), respectively. The pharmacokinetic studies showed that the SSa-SSd-Lip had increased circulation time, decreased Cl, and increased AUC, MRT and T1/2ß (p < 0.05) for both SSa and SSd after intravenous administration in comparison with solution.

[Pharmacokinetic study on combined application of gastrodin and puerarin in rats].

To establish a HPLC method for simultaneously determining plasma concentrations of gastrodin (Gas) and its metabolites hydroxybenzyl alcohol (HBA), puerarin (Pur) and internal standard (IS) p-hydroxyphenylethanol (Tyr) in rats and studying the pharmacokinetic process and interactions of gastrodin and puerarin after single and combined intravenous injection and oral administration. With Tyr as the internal standard, plasma samples were processed with methanol for protein precipitation, supernatant was dried with N2, and residues were re-dissolved with acetonitrile-0.05% phosphoric acid (20: 80). Chromatography was carried out on an Agilent ZORBAX SB-Aq C18 column (4.6 mm x 250 mm, 5 µm), with acetonitrile-0.05% phosphoric acid as the gradient mobile phase for the gradient elution. The UV detector wavelength was set at 221 nm for Gas HBA and IS and 250 nm for Pur. After the single or combined administration of Gas and Pur, their plasma concentrations in rats were detected. WinNonlin 5.2 pharmacokinetic software and SPSS 17. 0 software were used to respectively calculate pharmacokinetic parameters of each group, make a statistical analysis and compare the pharmacokinetic processes of Gas and Pur after the single or combined administration. According to the results, the absolute recoveries between low, media and high concentrations of Gas, HBA and Pur and IS as well as Tyr were more than 77.20%, with a good linearity (r > 0.999 6, n = 5) for Gas, HBA and Pur within concentration ranges of 0.10-101, 0.03-7.58 and 0.05-5.98 mg xL ('1) respectively. The lower limits of quantification for Gas, HBA and Pur were 0.10, 0.03, 0.05 mg x L(-1), respectively. Their in-ra-day and inter-day precisions were less than 12% with the accuracy between 85. 1% -1 10. %. All of the three substances and IS were stable during the whole analysis process. The findings showed significant differences in the main in vivo pharmacokinetic parame-ers in rats (AUC, C.(max) T,½ T.(max) MRT) after the single and combined administration of Gas and Pur. Either after the oral adminis-ration or after the intravenous injection, parameters showed a lower clearance rate ( L) longer mean residence time ( RT) and higher relative bioavailability, especially after the oral administration. Specifically, the relative bioavailability of the combined oral ad-inistration of Pur was 10. 7 times of that of the single administration, while that of Gas was 1. times of that of the single administra-ion. The combined administration of Gas and Pur can promote the absorption, decrease the elimination rate and prolong the mean resi-ence time. The method is simple and accurate and can be applied in the simultaneous determination of plasma concentrations of Gas, HBA and Pur in rats and the pharmacokinetic studies.

[Investigation of characteristic microstructures of adhesive interface in wood/bamboo composite material by synchrotron radiation X-ray phase contrast microscopy].

Third-generation synchrotron radiation X-ray phase-contrast microscopy(XPCM)can be used for obtaining image with edge enhancement, and achieve the high contrast imaging of low-Z materials with the spatial coherence peculiarity of X-rays. In the present paper, the characteristic microstructures of adhesive at the interface and their penetration in wood/bamboo composite material were investigated systematically by XPCM at Shanghai Synchrotron Radiation Facility (SSRF). And the effect of several processing techniques was analyzed for the adhesive penetration in wood/bamboo materials. The results show that the synchrotron radiation XPCM is expected to be one of the important precision detection methods for wood-based panels.

Enhancing Hot Air Drying Efficiency through Electrostatic Field-Ultrasonic Coupling Pretreatment.

The drying of compact and biologically active materials presents significant challenges. In this study, we propose using electrostatic field-ultrasonic coupling pretreatment to enhance the drying efficiency of ginkgo fruits. We designed and constructed an experimental device to investigate the effects of ultrasonic power, pretreatment time, hot air drying temperature, and electrostatic field voltage on the moisture content of the fruits. We used the response surface methodology to identify optimal process conditions and further explored the kinetic model for the moisture content of the fruits under the pretreatment. The results showed that the optimal process parameters for electrostatic-ultrasound pretreatment and the drying of ginkgo fruits were: an electrostatic field voltage of 11.252 kV, an ultrasound power of 590.074 W, a pretreatment time of 32.799 min, and a hot air drying temperature of 85 °C. Under the optimized process conditions, the correlation between the moisture content of ginkgo fruits and the two-term drying kinetics model was the highest. After electrostatic-ultrasound coupling pretreatment, the drying rate of ginkgo fruits was significantly improved during hot air drying.

Dietary and Sexual Correlates of Gut Microbiota in the Japanese Gecko, Gekko japonicus (Schlegel, 1836).

Numerous studies have demonstrated that multiple intrinsic and extrinsic factors shape the structure and composition of gut microbiota in a host. The disorder of the gut microbiota may trigger various host diseases. Here, we collected fecal samples from wild-caught Japanese geckos (Gekko japonicus) and captive conspecifics fed with mealworms (mealworm-fed geckos) and fruit flies (fly-fed geckos), aiming to examine the dietary and sexual correlates of the gut microbiota. We used 16S rRNA gene sequencing technology to determine the composition of the gut microbiota. The dominant phyla with a mean relative abundance higher than 10% were Verrucomicrobiota, Bacteroidota, and Firmicutes. Gut microbial community richness and diversity were higher in mealworm-fed geckos than in wild geckos. Neither community evenness nor beta diversity of gut microbiota differed among wild, mealworm-fed, and fly-fed geckos. The beta rather than alpha diversity of gut microbiota was sex dependent. Based on the relative abundance of gut bacteria and their gene functions, we concluded that gut microbiota contributed more significantly to the host's metabolic and immune functions. A higher diversity of gut microbiota in mealworm-fed geckos could result from higher chitin content in insects of the order Coleoptera. This study not only provides basic information about the gut microbiota of G. japonicus but also shows that gut microbiota correlates with dietary habits and sex in the species.

Population-based study on the prevalence of and risk factors for human papillomavirus infection in Qujing of Yunnan province, Southwest China.

BACKGROUND: Human papillomavirus (HPV) infection causes cervical cancer and premalignant lesions of the cervix. Prevalence of HPV infection and HPV genotypes vary among different regions. However there is no data on the prevalence of HPV infection and HPV genotypes from southwest China. This study was undertaken to determine the prevalence of and risk factors for HR-HPV infection in Qujing of Yunnan province, southwest China to provide comprehensive baseline data for future screening strategies. METHODS: A sample of 5936 women was chosen by the multi-stage stratified cluster sampling method with selection probabilities proportional to size (PPS). An epidemiological questionnaire was conducted via a face-to-face interview and cervical specimens were taken for HPV DNA testing by Digene Hybrid Capture 2 (HC2) test. HPV Genotyping Reverse Hybridization Test was used for HPV genotyping. Proportions were compared by Chi-squared tests, and logistic regression was utilized to evaluate risk factors. RESULTS: The median age was 38 years and the inter-quartile range was from 31 years to 47 years. 97.3% of the study population was Han nationality. Overall prevalence of HR-HPV infection was 8.3% (494/5936) and bimodal age distribution of HPV infection was observed. The five most prevalent HR-HPV genotypes were HPV-16(3.4%), HPV-56(1.7%), HPV-58(1.4%), HPV-33(1.2%) and HPV-52(0.88%). Multiple HPV infections were identified in 50.5% (208/412) of the positive genotyping specimens. Multivariate logistic regression model indicated that parity (OR = 1.35, 95% CI: 1.18-1.53, p < 0.0001) was a risk factor for HR-HPV infection, and age of 50-65 years (OR = 0.60, 95% CI: 0.45-0.80, p = 0.0005), being married or in stable relationship (OR = 0.55, 95% CI: 0.31-0.96, p = 0.035) were protective factors. CONCLUSIONS: This study provided baseline data on HR-HPV prevalence in the general female population in Qujing of Yunnan province, southwest China. The finding of multiple HPV infections and bimodal age distribution revealed that HPV screening is necessary for perimenopausal women in future.

[Distribution of lignin in Chinese fir branches determined by ultraviolet microspectrometer].

Distribution of lignin in the cell walls of Chinese fir branches emerged in the spring season were first studied by using ultraviolet microscope based on their cell microstructure observation and lignin qualitative measurement by the lightmicroscope and confocal laser scanning microscope. The results showed that the contents of lignin are inhomogeneously distributed in different micro-areas of the cell walls. The order of lignin concentrations is the cell corner>the middle lamellar>the secondary with the absorbance values of ultraviolet wave of 0.489, 0.307 and 0.278, respectively. The result of quantitative analysis consists with that of qualitative analysis. A new measurement method was proposed to study the distribution of lignin content in wood cell walls in CFhina.

Prediction of factors influencing the timing and prognosis of early tracheostomy in patients with multiple rib fractures: A propensity score matching analysis.

Objective: To investigate the factors affecting the timing and prognosis of early tracheostomy in multiple rib fracture patients. Methods: A retrospective case-control study was used to analyze the clinical data of 222 patients with multiple rib fractures who underwent tracheotomy in the Affiliated Hospital of Yangzhou University from February 2015 to October 2021. According to the time from tracheal intubation to tracheostomy after admission, the patients were divided into two groups: the early tracheostomy group (within 7 days after tracheal intubation, ET) and late tracheostomy group (after the 7th day, LT). Propensity score matching (PSM) was used to eliminate the differences in baseline characteristics Logistic regression was used to predict the independent risk factors for early tracheostomy. Kaplan-Meier and Cox survival analyses were used to analyze the influencing factors of the 28-day survival. Results: According to the propensity score matching analysis, a total of 174 patients were finally included in the study. Among them, there were 87 patients in the ET group and 87 patients in the LT group. After propensity score matching, Number of total rib fractures (NTRF) (P < 0.001), Acute respiratory distress syndrome (ARDS) (P < 0.001) and Volume of pulmonary contusion(VPC) (P < 0.000) in the ET group were higher than those in the LT group. Univariate analysis showed that the patients who underwent ET had a higher survival rate than those who underwent LT (P = 0.021). Pearson's analysis showed that there was a significant correlation between NTRF and VPC (r = 0.369, P = 0.001). A receiver operating characteristic(ROC)curve analysis showed that the areas under the curves were 0.832 and 0.804. The best cutoff-value values of the VPC and NTRF were 23.9 and 8.5, respectively. The Cox survival analysis showed that the timing of tracheostomy (HR = 2.51 95% CI, 1.12-5.57, P = 0.004) and age (HR = 1.53 95% CI, 1.00-2.05, P = 0.042) of the patients had a significant impact on the 28-day survival of patients with multiple rib fractures. In addition, The Kaplan-Meier survival analysis showed that the 28-day survival of patients in the ET group was significantly better than that of the LT group, P = 0.01. Conclusions: NTRF, ADRS and VPC are independent risk factors for the timing and prognosis of early tracheotomy. A VPC ≥ 23.9% and/or an NTRF ≥ 8.5 could be used as predictors of ET in patients with multiple rib fractures. Predicting the timing of early tracheostomy also need prediction models in the future.

TNFα-induced IP3R1 expression through TNFR1/PC-PLC/PKCα and TNFR2 signalling pathways in human mesangial cell.

BACKGROUND: Little information is available regarding the mechanisms involved in cytokine-induced type 1 inositol 1,4,5-trisphosphate receptor (IP(3)R1) expression in human mesangial cells (HMCs) in the occurrence of hepatorenal syndrome (HRS). Over-expression of IP(3)R1 would enhance both IP(3)-binding activity and sensitivity. We hypothesize that it is possible that increased IP(3)R1, induced by TNFα, would lead to increased IP(3) sensitivity in response to a variety of vasoconstrictors, and promote HMC contraction and thus lead to reduced GFP, promoting HRS occurrence and development. METHODS: Quantitative real-time polymerase chain reaction and immunoblot assay were used to examine the effects of TNFα on IP(3)R1 mRNA and protein expression. Several inhibitors of kinases, depletion PKC, over-expression of dominant-negative mutant of PKC and non-radioactive PKC assay were used to examine the mechanism of signal transduction of TNFα-regulated IP(3)R1 in HMCs. RESULTS: TNFα increased IP(3)R1 mRNA and protein expression in HMCs, an effect that was blocked by prolonged incubated chronic PMA, D609, safingol and also by transfection with domain-negative PKCα construct. TNFα activated and promoted autophosphorylation of the PKCα. In addition, both anti-TNFR1 and anti-TNFR2 antibodies blocked TNFα-induced IP(3)R1 protein expression, while only anti-TNFR1 antibodies but not anti-TNFR2 antibodies attenuated TNFα-induced PKCα activity. CONCLUSIONS: TNFα increased the expression of IP(3)R1, and this was mediated, at least in part, through the TNFR1/PC-PLC/PKCα and TNFR2 signalling pathways in HMCs.

A simple RP-HPLC method for the simultaneous determination of curcumin and its prodrug, curcumin didecanoate, in rat plasma and the application to pharmacokinetic study.

A high-performance liquid chromatographic (HPLC) method has been developed for the simultaneous determination of curcumin and its prodrug, curcumin didecanoate (CurDD), in rat plasma. The analytes were extracted by ethyl acetate following the addition of sodium dodecyl sulfate, and separated on a reverse-phase C(18) column using a gradient mobile phase system of acetonitrile-tetrahydrofuran-water containing 0.1% formic acid. Detection by UV absorption at 425 nm gave a lower limit of quantitation (LLOQ) of 5 and 10 ng/mL for curcumin and CurDD in 50 µL of plasma, respectively. Intra- and inter-day precisions of quality control samples except those at LLOQ were within 15% for curcumin and CurDD, respectively, and the accuracies for both compounds were between 93.9 and 108%. The method was successfully applied to determine plasma concentration-time curves of curcumin and CurDD in rats following intravenous (i.v.) administration of curcumin or CurDD at doses of 1 mg/kg (calculated as curcumin). The results suggested that i.v. dosed CurDD provided sustained plasma levels of curcumin.

[Effect of sequential method of nourishing yin and tonifying yang on serum sex hormone levels in rats with polycystic ovary syndrome].

OBJECTIVE: To observe the effect of sequential method of nourishing yin and tonifying yang on serum sex hormone levels in rats with polycystic ovary syndrome (PCOS), and to explore its possible mechanisms. METHODS: PCOS rat models were established by injecting dehydroepiandrosteron (DHEA) subcutaneously. A total of 70 21-day female SD rats were randomly divided into seven groups, ten in each group: the normal control, the model control, the yin nourishing, the yang tonifying, the sequential method, the Westem medicine, and the combined medication. Rats in all groups were treated by gastrogavage for twelve successive days. Serum testosterone (T) and estradiol (E2) were detected by chemoluminescence method. Serum androstenedione (A), follicle stimulating hormone (FSH), luteinizing hormone (LH), insulin (INS), and insulin-like growth factor-I (IGF-I) were detected by radioimmunoassay (RIA). RESULTS: Serum levels of A, T, and E2 were significantly higher in the model control group than in the normal control group (P < 0.05, P < 0.01). Serum levels of A and T were obviously lower in the sequential group than in the model control group with significant difference (P < 0.01). The serum E2 level was obviously lower in the sequential group than in the model control group (P < 0.05). Serum levels of INS and IGF-I were significantly higher in the model control group than in the normal control group (P < 0.05). Serum levels of INS and IGF-I were significantly lower in the sequential group than in the model control group (P < 0.01). CONCLUSIONS: The sequential method of nourishing yin and tonifying yang can lower serum levels of A, T, E2, INS, and IGF-I, and reverse its endocrine disturbance in PCOS rats. One of its mechanisms in decreasing serum levels of A, T, E2 might be possibly through lowering serum levels of INS and IGF-I.