OBJECTIVETo investigate the possible relationship between the analgesic effect of acupuncture and connexin 43.

METHODSConnexin 43 gene knock-out mice were divided into 4 groups: a wide type (WT) control group, a WT acupuncture group, a heterozygous (HT) control group and HT acupuncture group. Hot-plate test and writhing response induced by acetic acid were used for investigating the analgesic effect of acupuncture.

RESULTSThere was no significant difference in the basic pain threshold value between HT and WT mice (P > 0.05). Acupuncture could significantly increase the pain threshold value, prolong the latency period of writhing body and decrease the number of writhing body as compared with pre-acupuncture in WT and HT mice (P < 0.01 or P < 0.05). The pain threshold, latency period of writhing and number of writhing body in HT mice were less than WT mice post-acupuncture (P < 0.05).

CONCLUSIONConnexin 43 gene knock-out might partially inhibit the analgesic effect of acupuncture, suggesting that connexin 43 is possibly related with meridians and the effect of acupuncture.

Acupuncture Analgesia ; Animals ; Connexin 43 ; genetics ; physiology ; Female ; Gap Junctions ; physiology ; Male ; Mice ; Pain Threshold

Adult ; Connexin 43 ; genetics ; Erythrokeratodermia Variabilis ; genetics ; Female ; Heterozygote ; Humans ; Male ; Mutation ; genetics ; Mutation, Missense ; genetics ; Pedigree

OBJECTIVETo study the mechanism involved in the control of glioma cell proliferation with transfection of connexin (Cx) 43 gene.

METHODSC6 rat glioma and TJ905 human glioblastoma cell lines without Cx43 gene expression were transfected with Cx43cDNA mediated by lipofectamine. Northern blot, in situ hybridization and immunohistochemical technology were used to detect the expression of Cx43mRNA and its protein with MTT assay and silver colloid stain for the detection of cell proliferation, TUNEL method for determination of cell apoptosis, scrape loading dye transfer (SLDT) for GJIC, Western blot and immunohistochemical technology for bFGF, PDGF, EGFR, IGF-I and IGFBP3 expression.

RESULTSCx 43 gene transfected glioma cells showed decreased proliferation, restored GJIC and decreased bFGF, PDGF, IGFBP3, except EGFR expression and cell apoptosis which showed no change.

CONCLUSIONThe mechanism of Cx 43 gene inhibiting gliomas cell proliferation is the restoration of GJIC and decreased autocrine growth factors.

Animals ; Apoptosis ; Cell Division ; physiology ; Connexin 43 ; genetics ; physiology ; DNA, Complementary ; genetics ; Glioma ; pathology ; Rats ; Transfection ; Tumor Cells, Cultured

OBJECTIVETo observe the differences of the expression of Cx43 in rats' acupoints and sham-acupoint when needled blocks, and explore the relatively specificity of acupoints.

METHODTwenty Wistar rats were randomly divided into two groups: an acupuncture group and a control group, ten in each group. The rats in the control group were not treated and sacrificed directly to by air embolism; the rats in the acupuncture group were needled at "Neiguan (PC 6)", "Zusanli (ST 36)", "Dazhui (GV 14)" and "Guanyuan (CV 4)", Sham-acupoints were on 1 cm next to each of the above acupoints and "Yongquan (KI 1)" were needled by hand manipulation, retented needles for 30 min, then all the rats were sacrificed by air embolism. With the rats fixed, the tissue blocks at the acupoints and sham-acupoints were collected, then quick-frozen and cut into sections. The sections were stained by immunohistochemical method (Cx43).

RESULTSThe expression of the rats' Cx43 at the acupoints was significantly higher than that at the non-points in both groups (all P < 0.01). The expression of Cx43 at the acupoints in the acupuncture group was significantly higher than that in the control group (all P < 0.01).

CONCLUSIONThe expression of Cx43 at acupoints and non-points is significantly different, which indicates that the acupoints are actually existed and have relatively specificity.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Connexin 43 ; genetics ; metabolism ; Gene Expression Regulation ; Humans ; Male ; Rats ; Rats, Wistar

To explore the role of connexin43 (Cx43) in gap junctional intercellular communication (GJIC) and propagated sensation along meridians, the expression of Cx43 in the rat epithelial cells and fibroblasts was studied both in vitro and in vivo. With the in vitro study, the rat epithelial cells and fibroblasts were cultured together, and the localization of Cx43 was detected by immunohistochemistry and indirect immunofluorescent cytochemistry and under confocal microscopy. And the expression of Cx43 on the surface of the cells was examined by flow cytometry. With the in vivo examination, 20 SD rats were randomized into control group (n = 10) and electrical acupuncture group (EA group, n = 10). EA ( 0.5-1. 5 V, 4-16 Hz , 30 min) was applied to "Zusanli" acupoint for 30 min at rat's hind paw, the localization of Cx43 was immunohistochemically detected. The immunohistochemical staining and indirect immunfluorescent cytochemistry showed that Cx43 was localized on the surface of the cells and in the cytoplasm. The relative expression level of Cx43 on the cellular membrane surfaces of the rat epithelial cells and fibroblasts, as determined by FACS, were 13.91% and 29.53% respectively. Our studied suggested that Cx43 might be involved in GJIC and propagated sensation along meridians.
Cells, Cultured ; Connexin 43/*biosynthesis ; Connexin 43/genetics ; Epithelial Cells/cytology ; Epithelial Cells/*metabolism ; Fibroblasts/cytology ; Fibroblasts/*metabolism ; Flow Cytometry ; Gap Junctions ; Meridians ; Microscopy, Confocal ; Random Allocation ; Rats, Sprague-Dawley

OBJECTIVETo explore the genetic basis for a patient with oculodentodigital dysplasia.

METHODSGenomic DNA was extracted from peripheral blood samples from the patient and his parents. Whole-exome sequencing was carried out for the trio family. Suspected mutation was verified by Sanger sequencing.

RESULTSA de novo c.412G>A mutation of the GJA1 gene was identified in the patient, which was validated by Sanger sequencing.

CONCLUSIONThe c.412G>A mutation of the GJA1 gene probably underlies the disease in the patient.

Adult ; Connexin 43 ; genetics ; Craniofacial Abnormalities ; genetics ; Exome ; Eye Abnormalities ; genetics ; Foot Deformities, Congenital ; genetics ; Humans ; Male ; Mutation ; Sequence Analysis, DNA ; Syndactyly ; genetics ; Tooth Abnormalities ; genetics

Connexin 43 ; genetics ; Connexins ; genetics ; Heart Defects, Congenital ; genetics ; Homeobox Protein Nkx-2.5 ; Homeodomain Proteins ; genetics ; Humans ; Infant ; Infant, Newborn ; Mutation ; genetics ; Myocardium ; metabolism ; Transcription Factors ; genetics

OBJECTIVETo explore action mechanism of electroacupuncture (EA) on treatment of Parkinson's disease (PD).

METHODSA total of 40 healthy male SD rats were randomly divided into a normal group, a sham operation group, a model group and an EA group, 10 rats in each group. PD rat model was duplicated by micro injection of 6-hydroxyl dopamine into right striatum of rats, and the rats in the sham operation group were treated with micro injection of 0. 9% NaCl. Rats in the normal group, model group and the sham operation group received no treatment; rats in the EA group were treated by EA at "Fengfu" (GV 16) and "Taichong" (LR 3) with continuous wave, 2 Hz in frequency, 1 mA in intensity for 30 min. The treatment was given once a day for total 2 weeks. Behavioral test was used to evaluate rotational behavior changes of PD rats. RT-PCR method was applied to detect the expression of GFAP (glial fiber acidic protein) mRNA and Cx43 (connexin 43) mRNA in the striatum.

RESULTSThe difference of rotational behavior was not significant before and after treatment in the model group (P>0. 05), while that in the EA group was significant (P<0. 01). The expression of GFAP mRNA and Cx43 mRNA in the model group was significantly higher than that in the normal group and sham operation group (all P<0. 01); after EA treatment, the expression of GFAP mRNA and Cx43 mRNA in the EA group was lower significantly than that in the model group (both P<0. 01).

CONCLUSIONThe action mechanism of EA for prevention and treatment of Parkinson' s disease may be associated with inhibiting the activation of astrocytes.

Acupuncture Points ; Animals ; Astrocytes ; Connexin 43 ; genetics ; metabolism ; Corpus Striatum ; metabolism ; Electroacupuncture ; Humans ; Male ; Nerve Tissue Proteins ; genetics ; metabolism ; Parkinson Disease ; genetics ; metabolism ; therapy ; Rats ; Rats, Sprague-Dawley

Animals ; Apoptosis ; Cell Cycle ; Cell Movement ; Cell Proliferation ; Connexin 43 ; genetics ; metabolism ; Down-Regulation ; Gap Junctions ; metabolism ; physiology ; Humans ; Neoplasms ; metabolism ; pathology ; Tumor Suppressor Proteins ; genetics ; metabolism

BACKGROUNDGap junction channels formed by connexin43 (Cx43) protein are important in cardiac morphogenesis, and Cx43 gene is thought to be associated with congenital heart malformation (CHM). This study was undertaken to detect the mutations of Cx43 in fetuses with CHM.

METHODSCx43 extron DNA was amplified by PCR from 16 fetuses with a variety of CHM. The PCR products were analyzed by SSCP and DNA sequencing. Thirty children who had no CHM were selected as controls.

RESULTSEight homozygous mutations of Cx43 were observed in a fetus with double outlet right ventricule (DORV), five of the 8 mutations were missense mutations including Arg239Trp, Ser251Thr, Ala253Pro, Pro283Leu and Thr290Asn, and the remaining 3 were silent polymorphisms including Gly252Gly, Pro256Pro and Thr275Thr. No mutations were found in other fetuses and the control group.

CONCLUSIONSMutations of Cx43 may be associated with congenital conotruncal anomalies. PCR-SSCP is an effective method for screening the mutations of Cx43.

Connexin 43 ; genetics ; Fetus ; metabolism ; Heart Defects, Congenital ; genetics ; Humans ; Mutation ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Sequence Analysis, DNA