[Preparation of Huoluo Xiaoling gel plaster and its transdermal penetration in vitro].

Huoluo Xiaoling Dan is a classical prescription commonly used for blood circulation and pain relief in clinic with obvious effects. To make it directly treat lesion and improve the effect, this research optimized the preparation process of Huoluo Xiaoling gel paste and further evaluated its in vitro transdermal absorption performance, so as to provide a scientific basis for its development and utilization. Using primary viscosity, holding viscosity, and sensory score as evaluation indexes, the matrix amount of gel paste was determined by the single factor test and Box-Behnken response surface method. The ultra-performance liquid chromatography(UPLC) method was established to determine the content of eight active ingredients, including Danshensu, ferulic acid, salvianolic acid B, salvianolic acid A, ligustilide, tanshinone Ⅱ_A, 11-keto-ß-boswellic(KBA), and 3-acetyl-11-keto-ß-boswellic acid(AKBA). A mo-dified Franz diffusion cell method was used to evaluate and compare the absorption properties of the gel paste without volatile oil and with volatile oil microemulsion. The results showed that the optimal prescription for Huoluo Xiaoling gel paste matrix was NP700(1.35 g), glycerol(7.00 g), micropowder silica gel(1.25 g), sodium carboxymethyl cellulose(0.20 g), tartaric acid(0.06 g), and glyceryl aluminum(0.04 g). The mass fractions of eight active ingredients in the paste were successively 0.48, 0.014, 0.95, 0.39, 0.57, 0.055, 0.35, and 0.97 mg·g~(-1). The results of the in vitro transdermal absorption test showed that the addition of the volatile oil or the volatile oil microemulsion promoted the transdermal absorption of the active ingredients, and the law of drug penetration conformed to the zero equation or the Higuchi equation. The gel paste prepared by the optimal prescription has good appearance and adhesion, with no residue, and has the characteristics of skeletal slow-release preparation, which is easy to reduce the number of administration, la-ying a foundation for the development of new external dosage forms of Huoluo Xiaoling Dan.

[Combination characteristics of frankincense and myrrh and progress and prospect of their combination efficacy and mechanism].

Herbal pair is formed based on the experience summary of doctors' deep understanding and perception of the medicinal nature in long-term clinical practice. It gradually becomes the exquisite structural unit for preparing traditional Chinese medicine(TCM) prescriptions, and often plays a core bridge role in the prescription combination. Frankincense and myrrh are raw resin materials of incense abroad, which are subsequently included as Chinese medicinal herbs and endowed with rich medicinal connotation. With the functions of relaxing Zang-fu organs, activating blood and relieving pain, they have definite clinical efficacy. From the perspective of herbal description and clinical application, this study systematically analyzed the combination of frankincense and myrrh as well as their combination proportion, efficacy characterization, diseases and syndromes, effective components and action mechanism. On this basis, the focus of in-depth research of frankincense-myrrh and the application prospects were proposed, in order to further reveal the potential meditation law of this herbal pair, thus contributing to clinical practice and drug innovation of traditional Chinese medicine, and providing reference for understanding of TCM medicinal nature and research of herbal pairs.

A Liquid Level Measurement Technique Outside a Sealed Metal Container Based on Ultrasonic Impedance and Echo Energy.

The proposed method for measuring the liquid level focuses on the ultrasonic impedance and echo energy inside a metal wall, to which the sensor is attached directly, not on ultrasonic waves that penetrate the gas-liquid medium of a container. Firstly, by analyzing the sound field distribution characteristics of the sensor in a metal wall, this paper proposes the concept of an "energy circle" and discusses how to calculate echo energy under three different states in detail. Meanwhile, an ultrasonic transmitting and receiving circuit is designed to convert the echo energy inside the energy circle into its equivalent electric power. Secondly, in order to find the two critical states of the energy circle in the process of liquid level detection, a program is designed to help with calculating two critical positions automatically. Finally, the proposed method is evaluated through a series of experiments, and the experimental results indicate that the proposed method is effective and accurate in calibration of the liquid level outside a sealed metal container.

Unexplained infertility patients have increased serum IL-2, IL-4, IL-6, IL-8, IL-21, TNFα, IFNγ and increased Tfh/CD4 T cell ratio: increased Tfh and IL-21 strongly correlate with presence of autoantibodies.

Infertility is a disease of the reproductive system that affects millions of people globally. Reproductive failure is a major medical issue adversely affecting human health in the 21st century. Many factors contribute to infertility, including immune conditions which may lead to immune infertility (immunologic infertility). It is known that specific T helper cells (Th) and their cytokines are involved in the stages of infertility. The aim of this study is to provide a new diagnostic approach to immunologic infertility by investigating the correlation of follicular helper T cells (Tfh) and their secreted cytokines with the autoantibodies in peripheral blood samples from immunologically infertile patients. Thirty (30) patients suffering from immune infertility and 20 control subjects were selected as the sample base for this study. The levels of Tfh, 20 cytokines and 4 antibodies were evaluated for this investigation and evaluated using flow cytometry, antibody chip and ELISA technologies. It was found that, in immunologically infertile patients, Tfh cell numbers were significantly higher than those in the control group. Likewise, seven (7) serum cytokines were expressed to a greater degree in infertile patients compared to the control group. Finally, four (4) antibodies were found to be higher in immunologically infertile patients. The results show that, among patients with immunologic infertility, the levels of Tfh cells and IL-21 were increased significantly in peripheral blood samples and correlate positively with the autoantibodies. IL-12 was positively correlated with the two antibodies, while TNFα was negatively correlated with two additional antibodies. The detection and quantification of Tfh cells, IL-21, IL-12 and TNFα may provide new diagnostic indicators to screen for immunologic infertility.

miR-206 regulates the growth of the teleost tilapia (Oreochromis niloticus) through the modulation of IGF-1 gene expression.

MicroRNAs (miRNAs) are ~22-nucleotide noncoding RNAs that play a crucial role in regulating muscle development. Our previous study shows that miR-206 is specifically expressed in tilapia skeletal muscle, and exhibits a dynamic expression pattern at different developmental stages. Here, we reveal that miR-206 emerges as a crucial regulator of tilapia growth. miR-206 loss of function leads to the acceleration of tilapia growth. IGF-1 is identified as the target gene of miR-206. miR-206 directly changes IGF-1 expression by targeting its 3' UTR, and inhibition of miR-206 substantially increases the IGF-1 mRNA level in vivo. Thus, miR-206 could be developed as a molecular marker to assist fish breeding.

miR-203b: a novel regulator of MyoD expression in tilapia skeletal muscle.

MyoD is one of the helix-loop-helix proteins regulating muscle-specific gene expression in tilapia. Tight regulation of the MyoD protein level is necessary for the precise regulation of skeletal muscle development. MicroRNAs (miRNAs) are a class of regulatory RNAs that post-transcriptionally regulate gene expression. An increasing amount of evidence has suggested that miRNAs play an important role in regulating skeletal muscle development. We reasoned that MyoD expression may be regulated by miRNAs. Predictions from bioinformatics have identified a putative miR-203b target site in the 3'-UTR of the MyoD gene. Interestingly, miR-203b expression is negatively correlated with MyoD expression, whereas miR-203b suppression leads to a significant increase in MyoD expression, thereby activating MyoD downstream genes. A 3'-UTR luciferase reporter assay further verifies the direct interaction between miR-203b and MyoD. Taken together, our results reveal a novel molecular mechanism in which miRNA participates in transcriptional circuits that regulate gene expression in tilapia skeletal muscle.

[Diagnostic value of thyroid microcarcinoma with a taller-than-wide shape in thyroid nodules].

OBJECTIVE: To explore the diagnostic value of thyroid microcarcinoma with a taller-than-wide shape (A/T ≥ 1) in transverse ultrasonographic planes. METHODS: There were a total of 683 thyroid nodules in 491 patients undergoing consecutive ultrasonography and surgery at our hospital between 2011 and 2012. The ultrasonography examinations of thyroid nodules were routinely performed in the transverse planes. A taller-than-wide shape was defined as a ratio of anteroposterior diameter to transverse diameter in transverse plane ≥ 1. The sensitivity and specificity for thyroid carcinoma were analyzed. The data of thyroid carcinoma were formulated as follows: group 1, nodules > 1 centimeter; group 2, ≤ 1 centimeter. Their ratios of A/T were compared. The data were analyzed with SPSS 17.0 statistical software package. RESULTS: Among 683 nodules, 499 were malignant and 184 benign. The sensitivity and specificity for malignancy were 61.5% and 84.2% respectively. There was significant difference (P = 0.000). A taller-than-wide shape (A/T ≥ 1) was more frequent in criterion 2 than criterion 1 (χ(2) = 4.380, P = 0.036). CONCLUSIONS: A taller-than-wide shape (A/T ≥ 1) is associated with thyroid malignancy especially thyroid microcarcinoma. And it is a useful ultrasonic feature for predicting thyroid microcarcinoma.

miR-429 regulation of osmotic stress transcription factor 1 (OSTF1) in tilapia during osmotic stress.

The Nile tilapia represents an excellent model for osmoregulation study. Osmotic stress transcription factor 1 (OSTF1) identified in tilapia gill epithelium is a critical element of osmosensory signal transduction by means of transcriptional regulation. Thus, tight regulation of OSTF1 level is necessary for tilapia osmotic adaptation. microRNAs (miRNAs), have emerged as a crucial regulator of gene expression at post-transcriptional level. We reasoned that OSTF1 expression could be regulated by miRNAs. By bioinformatics analysis, we identified a putative miR-429 binding site in the OSTF1 mRNA. Interestingly, miR-429 is down-regulated in tilapia upon osmotic stress, consistent with OSTF1 protein up-regulation. miR-429 directly regulates OSTF1 expression by targeting its 3'-UTR, and inhibition of miR-429 substantially increases OSTF1 level in vivo. Moreover, miR-429 loss of function could influence the regulation of plasma osmolality and ion concentration responding to osmotic stress. Taken together, miR-429 is an endogenous regulator of OSTF1 expression, which participates in a regulatory circuit that allows rapid gene program transitions in response to osmotic stress.

MiR-30c: a novel regulator of salt tolerance in tilapia.

miRNAs comprise a class of ~22 nt noncoding RNAs that modulate the stability and/or translational potential of their mRNA targets. Emerging data suggest that stress conditions can alter the biogenesis of miRNAs, thereby changing the expression of mRNA targets. Here, we reveal that miR-30c, a kidney-enriched miRNA, emerges as a crucial osmoregulator in Nile tilapia. miR-30c loss of function leads to an inability to respond to osmotic stress. We identify HSP70 as one of the direct regulatory targets of miR-30c. miR-30c directly regulates HSP70 by targeting its 3'-UTR, and inhibition of miR-30c substantially increases HSP70 mRNA level in vivo. Taken together, our experiments suggest that miRNAs participate in a regulatory circuit that allows rapid gene program transitions in response to osmotic stress. miR-30c may be developed as a molecular marker to assist to breed or genetically engineer salt tolerant species.

Potential roles of gut microbiota and microbial metabolites in chronic inflammatory pain and the mechanisms of therapy drugs.

Observational findings achieved that gut microbes mediate human metabolic health and disease risk. The types of intestinal microorganisms depend on the intake of food and drugs and are also related to their metabolic level and genetic factors. Recent studies have shown that chronic inflammatory pain is closely related to intestinal microbial homeostasis. Compared with the normal intestinal flora, the composition of intestinal flora in patients with chronic inflammatory pain had significant changes in Actinomycetes, Firmicutes, Bacteroidetes, etc. At the same time, short-chain fatty acids and amino acids, the metabolites of intestinal microorganisms, can regulate neural signal molecules and signaling pathways, thus affecting the development trend of chronic inflammatory pain. Glucocorticoids and non-steroidal anti-inflammatory drugs in the treatment of chronic inflammatory pain, the main mechanism is to affect the secretion of inflammatory factors and the abundance of intestinal bacteria. This article reviews the relationship between intestinal microorganisms and their metabolites on chronic inflammatory pain and the possible mechanism.

Bear bile powder ameliorates type 2 diabetes via modulation of metabolic profiles, gut microbiota, and metabolites.

Introduction: Bear bile powder (BBP) is widely used in the clinic and has a hypoglycemic effect, but its mechanism is not clear. Methods: In this study, type 2 diabetes mellitus (T2DM) rats induced by a high-sugar and high-fat diet combined with streptozotocin were given BBP, and biochemical indexes, pathological sections, metabonomics, intestinal microbiota (IM) and short-chain fatty acids (SCFAs) were determined. Results: The results showed that BBP could reduce blood glucose, relieve inflammation, insulin resistance, and lipid metabolism disorder, and alleviate tissue damage of the liver, spleen, kidney, and pancreas in T2DM rats. It is worth noting that BBP can reverse the changes in blood and urine metabolites in T2DM rats, which are mainly related to tryptophan metabolism, pentose and glucuronate interconversions, starch and sucrose metabolism, and glycerophospholipid metabolism. In addition, BBP restored IM disorder in T2DM rats, decreased the abundance of Allobaculum, Blautia, Dubosiella, and Anaerostipes, enriched the abundance of Lactobacillus, Romboutsia, UCG-005, and norank_f__Eggerthellaceae, and increased the concentration of SCFAs in intestinal contents. Discussion: These findings suggest that BBP may improve T2DM by regulating multiple metabolic pathways, IM composition, and SCFAs levels.

The time interval between hCG priming and oocyte retrieval in ART program: a meta-analysis.

OBJECTIVE: To evaluate the relationship between different hCG priming-to-oocyte retrieval intervals and assisted reproductive technology (ART) outcome. METHODS: We systematically searched PubMed, EMBASE, the Cochrane Library, Science Citation Index, Chinese biomedicine (CBM) literature database, and Chinese Journal Full-text Database for randomized controlled trials (RCTs) published up to November 2010. Data was extracted from the studies by two independent reviewers. Statistical analysis was performed with Cochrane Collaboration's Review Manager (RevMan) 5.0.2. From extracted data, Risk Ratio (RR) with 95% confidence interval (CI) was calculated. RESULTS: 5 RCTs totaling 895 participants were included. Oocyte maturation rate was higher in the long interval group compared with short interval group (RR, 0.67; 95% CI, 0.62-0.73). There were no significant difference between the two groups with regard to fertilization rate (RR, 0.99; 95% CI, 0.94-1.04), implantation rate (RR, 0.91; 95% CI, 0.40-2.04), and pregnancy rate (RR, 0.79; 95% CI, 0.58-1.08). CONCLUSION: The percentage of mature (MII) oocytes can be increased by prolonging the interval between hCG priming and oocyte retrieval. The prolonged interval could not increase the fertilization rate, implantation rate, and pregnancy rate. Although there was evidence to confirm the results, they still need to be confirmed by large-sample, multicenter, randomized controlled trials. The time interval dependent mechanisms responsible for ART performance need to be elucidated.

Systemic evaluation of hypoxic-ischemic brain injury in neonatal rats.

The objective of the study was to evaluate the systematically rat model of neonatal hypoxic-ischemic brain damage. The right carotid arteries of 7-day-old healthy Wistar rats were ligated, and then, the rats were subjected to an environment with 8 % of oxygen. Four weeks after the birth, neurobehavioral test, water maze test, and motor-evoked potential and neuropathologic examinations were performed. The footprint analysis showed significantly larger and instable paces in the hypoxic-ischemic group (P < 0.05); the time that rats crossed the balance beam in the hypoxic-ischemic group was longer than the control group (P < 0.05). The water maze test showed that the escape latency of hypoxic-ischemic group was significantly longer than that of control group (P < 0.05). The hindlimb quadriceps compound muscle-evoked potential CMEP of rats in hypoxic-ischemic group showed that the wave amplitude was lower than that of control group (P < 0.05). HE staining showed visible periventricular leukomalacia in hypoxic-ischemic groups; disrupted nuclear membrane was detected in the IH group with transelectronmicroscopy; Immunohistochemistry: compared with control group, MBP-positive neurocytes decreased, glial fibrillary acidic protein positive neurocytes increased in the periventricular zone (P < 0.05). Carotid artery ligation combining the hypoxic chamber created a reliable and stable rat model of neonatal hypoxic-ischemic brain damage and can be used for experimental research related to management of cerebral palsy.

Application of Real-time Ultrasound Elastography in Diagnosing Benign and Malignant Thyroid Solid Nodules.

OBJECTIVE: Real-time ultrasound elastography (US-E) is a helpful tool in diagnosing thyroid nodules. This study aims to evaluate thyroid solid nodules, to establish the accuracy of US-E in providing information on the nature of these nodules, and to assess the clinical value of elasticity scores (ES) and strain ratio (SR) in differentiating thyroid solid nodules and to explore its distribution characteristics using pathological analysis as reference. METHODS: Traditional ultrasonography and US-E were performed on 131 thyroid solid nodules (99 benign ones and 32 malignant ones) in 120 patients (78 females and 41 males). Three radiologists evaluated the nodules based on a four-degree elasticity scoring system. The nodules were classified according to the ES as soft (ES 1-2) or hard (ES 3-4). The SR was calculated online. RESULTS: The sensitivity and specificity of the ES for thyroid cancer diagnosis were 78% and 80%, respectively. SR values ≥ 2.9 used as a standard to distinguish benign from malignant nodules had a sensitivity of 87% and a specificity of 92%. The SR of the benign lesions was 1.64±1.37, which was significantly different from that of malignant lesions, which was 4.96±2.13 (P<0.01). CONCLUSIONS: Both the ES and SR were higher in malignant nodules than those in benign ones. Real-time US-E was a useful index in the differential diagnosis of thyroid solid nodules. It can provide quantitative information on thyroid nodule characterization and improve diagnostic confidence.

[Studies on antigencity of human immunodeficiency virus type 1 (HIV-1) external glycoprotein as well as its expression in Pichia pastoris].

Based on the computer simulation, we analyzed hydrophobicity, potential epitope of recombined subtypes HIV-1 Env protein (851 amino acids) from Guangxi in China. Compared with conservative peptides of other subtypes in env protein, three sequences (469-511aa, 538-674aa, 700-734aa) were selected to recombine into a chimeric gene that codes three conservative epitope peptides with stronger antigencity, and was constructed in the yeast expression plasmid pPICZB. Chimeric proteins were expressed in Pichia pastoris under the induction of methanol, and were analyzed by SDS-PAGE and Westernblot. The results showed that fusion proteins of three-segment antigen were expressed in Pichia pastoris and that specific protein band at the site of 40kD was target protein, which is interacted with HIV-1 serum. The target proteins were purified by metal Ni-sepharose 4B, and were demonstrated to possess good antigenic specificity from the data of ELISA. This chimeric antigen may be used as research and developed into HIV diagnostic reagents.

BAF complex is closely related to and interacts with NF1/CTF and RNA polymerase II in gene transcriptional activation.

Brg- or hBrm-associated factor (BAF) complexes, a chromatin-remodeling complex family of mammalian cells, facilitate transcriptional activity by remodeling nucleosome structure. Brg1 is the core subunit of Brg-associated factor complexes. In the present study, we investigated the spatial relationship between Brg1 and nuclear factor 1 (NF1/CTF) and RNA polymerase II (RNAP II) upon gene transcriptional activation in vivo by employing immuno-gold labeling. The data showed that Brg1 was closely co-localized with NF1/CTF and RNAP II in HeLa cells. Moreover, the co-immunoprecipitation assay further revealed that Brg1 can be isolated together with NF1/CTF and RNAP II in the ConA-stimulated, but not the resting, T lymphocyte. The combined results suggested that BAF complexes can interact with NF1/CTF and RNAP II, and this interaction is closely dependent on the activation of gene transcription.