As a member of orphan G protein-coupled receptors (oGPCRs), hGPCRc was cloned from human colon tissue and analyzed by bioinformatic softwares. It was showed that the corresponding amino acids of hGPCRc formed seven-transmembrane domains as the key characteristic of GPCRs. Then, the recombinant GFP-hGPCRc was constructed by fussing hGPCRc into pEGFP-N1 carrying green fluorescent protein (GFP) gene, and CHO-K1 cells were subsequently transfected with the GFP-hGPCRc or pEGFP-N1. The green fluorescence protein expression in the two different transfected cells was observed under the laser scanning confocal microscopy (LSCM). It was showed that green fluorescence protein was distributed in the whole bodies of the cells transfected with pEGFP-N1, but mainly distributed on the plasma membrane and cytoplasm membrane transfected with GFP-hGPCRc. Thus, the localization on the membrane of hGPCRc was accorded with the predication by bioinformatic analysis. The expression analysis of hGPCRc by RT-PCR indicated that hGPCRc was abundantly expressed in heart, kidney, cerebel and colon etc., but absent in liver, cerebra, small intestine and muscle etc. The expressing profile of hGPCRc could provide some useful clues to understanding its effects on embryonic development and physiological functions.
Amino Acid Sequence ; Animals ; CHO Cells ; Cell Membrane ; metabolism ; Cricetinae ; Cricetulus ; Gene Expression Profiling ; Green Fluorescent Proteins ; genetics ; Humans ; Molecular Sequence Data ; Receptors, G-Protein-Coupled ; genetics ; metabolism ; Tissue Distribution ; Transfection

OBJECTIVETo observe the images of early lesions of condylar cartilage of osteoarthritic rats in synchrotron radiation diffraction enhanced imaging (DEI).

METHODSThe animal model of temporomandibular joint osteoarthrosis was established in rat following the method of partial resection of the joint disc. The changes of osteoarthritic condylar cartilage in different pathological stages were observed by DEI and compared with those in situ histopathological sections.

RESULTSWith DEI, straight and orbicular lines were detected in condylar cartilage 45 to 60 days after discs resection. The lines were confirmed by histopathology to be collagen degradation and tiny fissure formation inside the cartilage.

CONCLUSIONSDEI is capable of imaging the early stages of pathological changes of excised condylar cartilage such as collagen degradation and tiny fissure formation, and this technique is of potential value to clinical application.

Animals ; Cartilage, Articular ; diagnostic imaging ; pathology ; Male ; Mandibular Condyle ; diagnostic imaging ; pathology ; Radiography ; Rats ; Rats, Wistar ; Temporomandibular Joint Disorders ; diagnostic imaging ; pathology ; X-Ray Diffraction ; methods

OBJECTIVEThis study sought to isolate and identify the proteins that interact with ataxin-3, to confirm the interacted domain, and to provide new clues for exploring the function of ataxin-3 and the pathogenesis of spinocerebellar ataxia type 3 and Machado-Joseph disease (SCA3/MJD).

METHODSYeast two-hybrid screen (MATCHMAKER GAL4 Two-Hybrid System 3) and regular molecular biologic techniques were undertaken to screen human brain cDNA library with mutant ataxin-3 bait. Two baits from both normal and mutant C-terminus of ataxin-3 were created by subcloned methods to determine which domain of ataxin-3 interacts with the putative associated proteins and to find out optimal candidate proteins that interact with C-terminus of ataxin-3. Confocal microscope was used to observe whether ataxin-3 co-localized with the obtained interacting proteins in mammalian cells.

RESULTSFive novel ataxin-3 interacting proteins were obtained, among which were three known proteins, namely human rhodopsin guanosine diphosphate dissociation inhibitor alpha, small ubiquitin-like modifier 1, and human neuronal amiloride-sensitive cation channel 2; the other two were unknown. Interacting domain analysis revealed that an unknown protein interacted with the C-terminus near the polyglutamine tract of ataxin-3, the other four all interacted with the N-terminus. In the nucleus of SH-SY5Y cell, small ubiquitin-like modifier 1 co-localized with the wild-type ataxin-3 and with the intranuclear aggregates formed by the mutant ataxin-3.

CONCLUSIONAn unknown protein probably interacting with C-terminus of ataxin-3 is firstly discovered, and the initiative findings suggest first that the interaction of small ubiquitin-like modifier 1 with N-terminus of ataxin-3 and the relevant sumoylation probably participate in the post-translation modifying of ataxin-3 and in the pathogenesis of SCA3/MJD.

Acid Sensing Ion Channels ; Ataxin-3 ; Cell Line, Tumor ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Confocal ; Mutation ; Nerve Tissue Proteins ; genetics ; metabolism ; Nuclear Proteins ; genetics ; metabolism ; Plasmids ; genetics ; Protein Binding ; Recombinant Fusion Proteins ; genetics ; metabolism ; Repressor Proteins ; genetics ; metabolism ; SUMO-1 Protein ; genetics ; metabolism ; Sodium Channels ; genetics ; metabolism ; Transfection ; Two-Hybrid System Techniques

OBJECTIVETo investigate the analgesic effects of Nourishing yin and Unblocking meridians Receipe (NUR) combined with opioid analgesics in managing cancer pain.

METHODSAll the patients enrolled were differentiated as of yin deficiency and meridian blocked syndrome type of TCM. Forty-one of them in the treated group were treated with NUR combined with opioid analgesics, while 43 of them in the control group were given opioid analgesics alone with successive 14 days as one treatment course for both groups.

RESULTSThe indexes of the treated group were superior to those in the control group as to the degree of pain-relieving, the therapeutic effect of analgesia, the occurrence frequency of cancer pain every day and its duration each time, the analgesic initial time, and the quality of life.

CONCLUSIONNUR combined with opioid analgesics in cancer pain management was more effective than opioid analgesics alone.

Adult ; Analgesics, Opioid ; administration & dosage ; adverse effects ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Female ; Humans ; Male ; Meridians ; Middle Aged ; Morphine ; administration & dosage ; adverse effects ; Neoplasms ; complications ; Pain ; drug therapy ; etiology ; Pain Measurement ; Treatment Outcome ; Yin Deficiency ; drug therapy ; Yin-Yang

OBJECTIVETo investigate the effect of solanine on the growth of human prostate cancer cell xenograft in nude mice.

METHODSHuman prostate cancer Du145 cells were injected into the subcutaneous layers on the back of nude mice. After a week, the mice bearing subcutaneous tumor graft were randomly divided into solanine treatment group and saline control group for treatment for 3 weeks. The tumor grafts were then harvested to evaluate the inhibition rate. The mRNA and protein expressions of cell cycle-related genes in the tumors were detected by qRT-PCR and Western blotting, respectively, and tumor cell apoptosis was detected using TUNEL method.

RESULTSThe tumor growth rate in solanine-treated group was significantly slower than that in the control group (P<0.01). The mRNA and protein expressions of C-myc, cyclin D1, cyclin E1, CDK2, CDK4 and CDK6 were significantly inhibited by solanine. Solanine significantly up-regulated p21 mRNA and protein expression in the tumors and induced a higher apoptosis rate of the tumor cells than saline (P<0.01).

CONCLUSIONThe tumor-inhibition effect of solanine is probably mediated by regulating the expressions of genes related with G1/S cell cycle arrest and cell apoptosis.

Animals ; Apoptosis ; Cyclin-Dependent Kinases ; metabolism ; Cyclins ; metabolism ; G1 Phase Cell Cycle Checkpoints ; Humans ; Male ; Mice ; Mice, Nude ; Neoplasm Transplantation ; pathology ; Prostatic Neoplasms ; drug therapy ; pathology ; S Phase ; Solanine ; pharmacology

Objective To investigate the clinical manifestations, diagnosis and treatment of primary intracranial tumors of multiple cellular origins. Methods The clinical data including the clinical manifestations, imaging findings, and treatment of 8 patients with primary intracranial tumors of multiple cellular origins were retrospectively analyzed, and the possible mechanism of these tumors were explored. Results A total of 16 primary intraeranial tumors were found in the 8 patients, including intraeranial meningioma (8 tumors), pituitary adenoma (3 tumors), schwannoma (2 tumors), and glioma (2 tumors). The tumors were resected in a single operation in 5 cases, and in two surgical sessions in 3 cases. Twelve tumors were radically resected, 3 were subtotally resected, and 1 was partially resected. Conclusion MR is an important diagnostic modality for primary intraeranial tumors of multiple cellular origins. Tumor resection in one or staged operations combined with radiotherapy or chemotherapy is the primary treatment option for these tumors.

OBJECTIVETo investigate the risk factors for the development of congenital anal atresia in neonates.

METHODSA total of 70 neonates who were admitted to 17 hospitals in Foshan, China from January 2011 to December 2014 were enrolled as case group, and another 70 neonates who were hospitalized during the same period and had no anal atresia or other severe deformities were enrolled as control group. Univariate and multivariate logistic regression analyses were used to investigate the risk factors for the development of congenital anal atresia.

RESULTSThe univariate analysis revealed that the age of mothers, presence of oral administration of folic acid, infection during early pregnancy, and polyhydramnios, and sex of neonates showed significant differences between the case and control groups (P<0.05). The multivariate logistic regression analysis revealed that infection during early pregnancy (OR=18.776) and male neonates (OR=9.304) were risk factors for congenital anal atresia, and oral administration of folic acid during early pregnancy was the protective factor (OR=0.086).

CONCLUSIONSInfection during early pregnancy is the risk factor for congenital anal atresia, and male neonates are more likely to develop congenital anal atresia than female neonates. Supplementation of folic acid during early pregnancy can reduce the risk of congenital anal atresia.

Anus, Imperforate ; etiology ; Female ; Humans ; Infant, Newborn ; Logistic Models ; Male ; Pregnancy ; Risk Factors

OBJECTIVETo investigate the levels of blood pressure and serum lipids, and examine the relationship between hypertension and hyperlipidemia in Hei Yi Zhuang Chinese living in Guangxi.

METHODSA total of 1056 people of Hei Yi Zhuang ethnicity were studied. Blood pressure, body height, body weight, and serum levels of lipids and apolipoprotein were measured. The data were compared with those in 925 people of Han ethnicity, who live in the same region.

RESULTSSystolic blood pressure and pulse pressure were significantly higher in Hei Yi Zhuang than Han Chinese (P < 0.001). The prevalence of isolated systolic hypertension and hypertension was also significantly higher in Hei Yi Zhuang than Han Chinese (P < 0.001). Serum concentrations of total cholesterol, triglycerides, low-density lipoprotein cholesterol, and apolipoprotein (Apo) B, and the prevalence of hypercholesterolemia and hyperlipidemia were significantly lower in Hei Yi Zhuang than Han Chinese (P < 0.05). Serum concentrations of high-density lipoprotein cholesterol and the Apo A1 to Apo B ratio were significantly higher in Hei Yi Zhuang than Han Chinese (P < 0.001). The prevalence of hypertension in Hei Yi Zhuang Chinese was positively associated with triglycerides (r = 0.425, P < 0.05), whereas the prevalence of hypertension in Han Chinese was positively correlated with total cholesterol (r = 0.623, P < 0.001).

CONCLUSIONThe present study revealed a significant difference in blood pressure and serum lipids between Hei Yi Zhuang and Han ethnic groups, and an association between hypertension and hyperlipidemia.

Adult ; Aged ; Asian Continental Ancestry Group ; ethnology ; Blood Pressure ; China ; epidemiology ; Female ; Humans ; Hyperlipidemias ; epidemiology ; ethnology ; Hypertension ; epidemiology ; ethnology ; Lipids ; blood ; Male ; Middle Aged ; Prevalence ; Sampling Studies ; Young Adult

Objective:To study the microbial community composition and diversity of brown-rot Gastrodia elata and its surface soil，in order to explain the relationship between brown-rot G. elata and soil microflora in G. elata planting process and provide theoretical basis for revealing the reasons of G. elata brown-rot disease. Method:Used internal transcribed spacer region（ITS） and 16S rDNA high-throughput sequencing technologies to detect the microbial diversity，community structure composition and community structure similarity of fungi and bacteria in healthy tuber，Brown-rot tuber，healthy soil and Brown-rot soil. Result:Compared with health groups，the number of fungi and bacteria operational taxonomic units（OTUs） was increased in brown-rot G. elata and its soil， and the abundance and diversity of fungi and bacteria in brown-rot G. elata soil were significantly decreased. The diversity of fungi in the tubers of brown-rot G. elata was significantly reduced，while the diversity of bacteria was significantly decreased. At the genus level， Mortierella was dominant fungi genus in healthy tuber and healthy soil，which was reduced 7.62% and 15.75% respectively in brown-rot tuber and brown-rot soil. And the dominant bacteria genus was Bradyrhizobium and Burkholderia-Paraburkholderia respectively. Ilyonectria was dominant fungi genus in brown-rot tuber and brown-rot soil，the dominant bacteria genus was Serratia and Bradyrhizobium respectively. Conclusion:The fungal flora in the tuber of brown-rot G. elata had a very high degree of similarity to that in the surrounding soil. These results indicated that the change of soil microbial fungal community caused the occurrence of G. elata brown-rot disease to a certain extent. And the pathogenic fungal Ilyonectria was dominant genus in fungi community of brown-rot tuber and brown-rot soil. Ilyonectria may have the main G. elata brown-rot disease pathogen.