Purpose To investigate the expression of muscarinic cholinergic receptor 3 (M3R) in small cell lung cancer (SCLC) and the correlation between it and clinical characteristics of SCLC. Methods The M3R expression was examined in 60 SCLC patients ad-mitted using immunohistochemistry. The patients were evaluated by clinical history, physical examination and chest CT. Survival time was got through telephone follow-up. Results The M3R expression in primary SCLC tissues was higher than that in normal lung tis-sues. The M3R expression had no difference between limited-disease and extensive-disease of SCLC. There was no correlation between M3R expression and age, gender, lymph node metastasis. The M3R expression was significantly correlated with smoking history. The survival time of M3R negative patients was longer than that of M3R positive patients in Kaplan-Meier survival curve. Conclusion The M3R expression in primary SCLC tissues was higher than that in normal lung tissues. M3R expression had some correlation with prog-nosis of SCLC patients.

OBJECTIVE:To provide reference for improving the registration success rate of new drug research and develop-ment(R&D)enterprises after adjusting drug registration system in China. METHODS:The national drug policies,regulations and related documents were comprehensively studied and combed to explore the main contents of drug registration system adjustment and its effects on the R&D forms,drug patent and on-site verification. The suggestions were put forward. RESULTS & CONCLU-SIONS:The adjusted drug management system showed new definitions for new drugs and generic drugs in China,as well as de-tailed requirements and regulations for drug R&D conditions,process and on-site inspection. It enhanced the protection of patents and patentees. The new drugs should be assessed its market value and clinical advantages. Data contents and requirements for drug registration declaration were made the relevant adjustments. New drug R&D enterprises should develop drug listed license manage-ment system as early as possible based on the implementation of listed license management system pilot,improve its efficiency and benefit. The new drug R&D should notice the clinical efficacy and demand of market,focus on the drugs with good clinical effica-cy in common diseases,frequently-occurring diseases and major diseases;and those for treating rare diseases,the elderly and chil-dren disease can use easy access or fast track to accelerate the speed of drug registration. New drug R&D enterprises should im-prove the R&D conditions,and standardize the management of development process to ensure the effective and smooth develop-ment.

Objective To investigate the expression of Toll-like receptor 4 (TLR4) and CD80 in unexplained recurrent spontaneous abortion and their relationship,discuss its possible roles in guiding eugenics,and provide a new idea for clinical diagnosis and prevention.Methods The study group were 60 cases with unexplained recurrent spontaneous abortion patients.The control group included 30 cases of normal early pregnancy without previous history of adverse pregnancy.The expressions of TLR4 and CD80 were detected with immunohistochemistry in the peripheral blood,chorion,and decidua from each case.Results TLR4 had higher expression in the peripheral blood,the chorion and decidua from the recurrent spontaneous abortion women than that from normal pregnancy women (P ＜ 0.05).Levels of CD80 were significantly higher in recurrent spontaneous abortion group than in normal pregnancy group (P ＜ 0.05).There was a positive correlation between TLR4 and CD80 (r =0.982,0.986,and 0.776,P ＜0.01).Conclusions The study shows that TLR4 and CD80 were expressed in peripheral blood lymphocytes and villi and decidua of early pregnancy (normal pregnancy or abnormal pregnancy).Higher expressions of TLR4 and CD80 might play an important role in the occurrence of recurrent spontaneous abortion.

OBJECTIVETo detect potential mutations in genes related with non-syndromic oculocutaneous albinism I-IV and ocular albinism type I in two couples who had given births to children with albinism.

METHODSAll exons of the non-syndromic albinism related genes TYR, OCA2, TYRP-1, MITF, SLC45A2 and GPR143 were subjected to deep sequencing. The results were verified with Sanger sequencing.

RESULTSFor the two female carriers, the coding region of the TYR gene was found to harbor a frameshift mutation c.925_926insC, which was also suspected to have been pathogenic. In one of the male partners, a nonsense mutations c.832C>T was found, which was also known to be pathogenic. Another male partner was found to harbor a TYR gene mutation c.346C>T, which was also known to be a pathogenic nonsense mutation.

CONCLUSIONThe coding region of the TYR gene c.925_926insC (p.Thr309ThrfsX9) probably underlies the OCA1 disease phenotype.

Adult ; Albinism, Oculocutaneous ; enzymology ; genetics ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Exons ; Female ; Frameshift Mutation ; Humans ; Male ; Membrane Glycoproteins ; genetics ; Molecular Sequence Data ; Mutation, Missense ; Oxidoreductases ; genetics ; Pedigree

Objective ToexploreamethodofusingCaspase-3smallinterferenceribonucleicacid (siRNA)in vitro to decrease the apoptosis in stable cell lines of bone marrow mesenchymal stem cell (MSC)in ordertoestablishMSCcelllineswithstableexpression.Methods VirusparticlescontainingCaspase-3siRNA were generated in 293FT cells by retroviral packaging system,then were transfected into MSC of rats. And the transfected cells were screened and cultured to get the stable MSC lines. According to the characteristics of retrovirus,the stable expression of the cell lines was identified by Western blot and real-time fluorescent quantitative PCR (RT-QPCR). The apoptosis of stable cell lines and normal MSC were detected by terminal dexynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL ) and their differences were compared.Results Comparedwiththenormalcells,thecaspase-3proteinexpressionandmRNAcontentof stable MSC lines were significantly reduced. In the same condition in vitro,the apoptosis quantity of stable MSC linessignificantlydecreasedcomparedwiththenormalcells.Conclusions StablecelllinesofMSCwithstable expression of Caspase-3 siRNA can be obtained by retroviral packaging system. The apoptosis quantity of stable MSC Lines significantly decreased compared with the normal cells.

The fungus Trichophyton schoenleinii (T. schoenleinii) is the causative agent of Trichophytosis and Tinea favosa of the scalp in certain regions of Eurasia and Africa. Human innate immune system plays an important role in combating with various pathogens including fungi. The inflammasome is one of the most critical arms of host innate immunity, which is a protein complex controlling maturation of IL-1β. To clarify whether T. schoenleinii is able to activate the inflammasome, we analyzed human monocytic cell line THP-1 for IL-1β production upon infection with T. schoenleinii strain isolated from Tinea favosa patients, and rapid IL-1β secretion from THP-1 cells was observed. Moreover, applying competitive inhibitors and gene specific silencing with shRNA, we found that T. schoenleinii induced IL-1β secretion, ASC pyroptosome formation as well as caspase-1 activation were all dependent on NLRP3. Cathepsin B activity, ROS production and K⁺ efflux were required for the inflammasome activation by T. schoenleinii. Our data thus reveal that the NLRP3 inflammasome plays an important role in host defense against T. schoenleinii, and suggest that manipulating NLRP3 signaling can be a novel approach for control of diseases caused by T. schoenleinii infection.
Animals ; Bone Marrow Cells ; cytology ; Carrier Proteins ; metabolism ; Caspase 1 ; metabolism ; Cell Line ; Dendritic Cells ; cytology ; metabolism ; microbiology ; Enzyme Activation ; Hot Temperature ; Humans ; Inflammasomes ; metabolism ; Interleukin-1beta ; biosynthesis ; metabolism ; Lysosomes ; metabolism ; Mice ; Monocytes ; cytology ; metabolism ; microbiology ; NLR Family, Pyrin Domain-Containing 3 Protein ; Potassium ; metabolism ; Reactive Oxygen Species ; metabolism ; Signal Transduction ; Trichophyton ; physiology

Objective: To explore the lipid-lowering effect and potential mechanism of the couplet medicines of Flastem Milkvetch Seed and Tribulus terrestris on hyperlipidemia rats. Methods: Forty-eight Sprague-Dawley rats were randomly divided into 6 groups, normal control group, high-fat model group, simvastatin group, Tribulus terrestris group, Flastem Milkvetch Seed group, and couplet medicines of Flastem Milkvetch Seed and Tribulus terrestris group, with 8 rats in each group. The normal control group was fed with basal diet, and the other groups were fed with high-fat diet to establish a hyperlipidemia rat model. At the same time, different group rats were treated with different drug respectively. 6 weeks later, the body weight and visceral index of the rats were measured. Levels of serum cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), and low density lipoprotein cholesterol (LDL-C) were detected by automatic biochemical analyzer. The mRNA and protein expression of HMG coenzyme A reductase (HMGCR), cholesterol 7-alpha hydroxy-lase (CYP7A1) and low density lipoprotein receptor (LDL-R) were detected by RT-PCR and western blot. Results: Compared with model group, the serum levels of TC, TG, and LDL-C were significantly decreased in all Chinese medicine groups, and the serum level of HDL-C was significantly increased in Flastem Milkvetch Seed group and couplet medicines of Flastem Milkvetch Seed and Tribulus terrestris group. Moreover, the mRNA and protein expression of HMGCR, CYP7A1, and LDL-R in rat liver also significantly increased in all Chinese medicine treatment groups. And couplet medicines of Flastem Milkvetch Seed and Tribulus terrestris group shows more significantly effect. Conclusion: Couplet medicines of Flastem Milkvetch Seed and Tribulus terrestris significantly regulate the serum lipid levels on hyperlipidemia rat model, and its mechanism may be related to the regulating expressions of HMGCR, CYP7A1, LDL-R gene.