Objective To investigate the prevalence of subclinical hypothyroidism and its relation to serum total cholesterol (TC),triglyceride (TG) levels and non-alcoholic fatty liver.Methods Serum levels of TC,TG, TSH,TT_3 and TT_4 were determined in 1 602 subjects which were divided into groups by sex and age (20-39,40- 59 and≥60 years).Fatty liver was diagnosed by type B ultrasonography.Restults ( 1 ) Prevalence of subclinical hypothyroidism was 6.6% in this population.(2) The mean levels of serum TC and TG in subclinical hypothyroidism group and normal group showed no significant difference ( both P＞0.05 ).(3) On the whole,the prevalences of subclinical hypothyroidism in hypercholesterolemia group and normal cholesterolemia group showed significant difference (P＜0.01 ).(4) In the three age groups of male and few.ale examinees,the prevalences of subclinical hypothyroidism in hypercholesterulemla group and normal cholesterolemia group showed no significant difference ( all P＞0.05 ).In the multiple regression analysis,TSH level was not related with raised TC level.(5) In the three age groups of male,there was no significant difference between the prevalences of subclinical hypothyroidism in hypertriglyceridemla group and normal triglyceridemia group ( all P＞0.05 ).In female,expect for the elderly group (＞60 years old ),the prevalence of subclinical hypothyroidism was higher in hypertriglyceridemia group than that in normal triglyceridemia.However,in the multiple regression analysis, increased TSH level was positively associated with increased serum TG level ( OR = 1.072,P=0.013 and OR = 1.102,P = 0.03 ).(6) The prevalence of subclinical hypothyroidism in non-alcobolic fatty liver group and normal group showed no significant difference ( P＞0.05 ).In the multiple regression analysis,TSH level was not the independent risk factor of non-alcoholic fatty liver (P＞0.05 ).Conclusion Subclinical hypothyroidism is not asseciated with serum TC level but positively associated with serum TG level.Subclinical hypothyroidism does not apparently increase the prevalence of non-alcoholic fatty liver.

Lonicerae Japonicae Flos was one of the most widely used traditional Chinese medicine for its special biological activities. The content of luteoloside, one of its major compounds, was an important standard for the quantity control of Lonicerae Japonicae Flos. The major method used for the detection of luteoloside was instrumental analysis. Compared with the ELISA method, instrumental analysis was time-consuming, complex pretreatment and low-throughout. Thus, it was significantly important to develop an enzyme-linked immunosorbent assay (ELISA) for luteoloside analysis. Here, the conjugates of luteoloside-bovine (LG-BSA) and luteoloside-ovalbumin (LG-OVA) were produced as the immunogen and coating antigen by the carbodiimide ( CDI) method, respectively. The conjugation ratio of carrier protein and the hapten in the conjugate were determined by UV-Vis spectrophotometry (UV). LG-BSA conjugate was used to immunize Bal b/c mice to produce antiserum. The titer and specificity of antiserum were detected by ELISA. The conjugation ratio of hapten and carries protein were 3. 7: 1 (LG-BSA) and 1. 0: 1 (LG-OVA). The antiserum titer was higher than 2 000 with the linear range of 18.4-4 852.4 μg x L(-1), R2 = 0.988 4 and IC50 = 298.7 μg x L(-1). The result showed that the conjugate antigen LG-BSA was synthesized successfully and the mice can produce specific antiserum injected with artificial antigen.
Animals ; Antibodies ; analysis ; immunology ; Antigens ; chemistry ; immunology ; Drugs, Chinese Herbal ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Immunization ; Lonicera ; chemistry ; immunology ; Mice ; Mice, Inbred BALB C

Objective To detect the mutations of ATP2C1 gene in patients with Hailey-Hailey dis- ease (HHD).Methods PCR and direct sequencing were performed in 17 patients and 120 healthy controls to screen the mutations in the exons of ATP2C1 gene.Results Eight mutations were identified in nine probands, including three deletion mutations (nt1464-1487 del/nt1462-1485del,1523delAT,2375delTTGT),three splice site mutations (360—2A→G,1415—2A→T,2243+2T→C) and two missence mutations (C920T and G1942T).None of the above mutations was found in the controls.Conclusion Eight specific novel mutations were identified in nine probands of HHD,which could be causative factors of the disease.

Objective To investigate the effects of dominant-negative truncation mutant?NTCF4, lacking the N-terminal form of TCF4 gene,on biological characteristics of renal cancer cell line GRC-I and explore the molecular mechanisms.Methods GRC-I cell was transfected with pCDNA3-?NTCF4 eukary- otie expression plasmid,pCDNA3 empty vector to construct the stable cell line GRC-I/?NTCF4 and GRC-I/ Mock respectively.The morphological changes of stable cells were observed and the cells growth curve was detected through light microscope.The cellular proliferation activities were determined using the MTT assay. The protein expression of Wnt pathway downstream target gene C-Myc and Cox-2 was evaluated by immuno- cytoehemieal method and Western Blot analysis.Results After the dominant-negative?NTCF4 gene was permanently expressed,the GRC-I/?NTCF4 stable cells morphologically showed that appearance changed from circular to long-spindle shape,growth rate decreased with less karyosehisis found,malignant pheno- types reversed to normal renal tubular cells.MTT assay revealed that the proliferation activities of GRC-1/?NTCF4 cells were inhibited by 11.2%-35.5% compared with GRC-I cells (P＜0.05),while the GRC- I/Mock cells have no difference with the control cells.Immunocytochemical analysis and Western Blot showed that the C-Myc and Cox-2 protein expression level of GRC-I/?ANTCF4 cells were significantly sup- pressed in comparison with that of GRC-I/Mock and GRC-I cells.Conclusions The dominant-negative truncation mutant?NTCF4 could partially inhibit the growth of renal cancer cells and down-regulate the pro- tein expression of Wnt pathway target gene C-Myc and Cox-2.These findings provide a experimental founda- tion for applying cell signal therapy to renal cell cancer by blocking the Wnt signaling pathway.

Objective To establish inductively coupled plasma mass spectrometry ( ICP -MS ) method for determining the concentration of ethaselen in human plasma , and to apply it to the pharmacokinetic study of ethaselen dispersible tablets.Methods Plasma samples were digested with nitric acid , then detected by ICP -MS method.The main pharmaco-kinetic parameters were calculated with non -compartmental analysis by Wi-nNonlin 5.2 software.Results The parameters pharmacokinetic results of ethaselen were as follows: tmax was ( 9.20 ± 0.98 ) h, Cmax was (597.58 ±221.73) ng· mL-1, AUC0-t was (2.57 ±0.92) ng· h· mL-1, the mean residence time (MRT) was (24.60 ±0.63)h.Conclusion The ICP-MS method is simple, rapid and sensitive, which is suitable for clini-cal determination of the concentration of ethaselen dispersible tablets in hu-man plasma.

OBJECTIVETo construct the cloning vector of glycoprotein G2 gene of hantavirus (HV), to analyze the sequence of G2 gene by the phylogenetic tree, and to study the differences among glycoprotein G2 genes from the world around.

METHODSEnvelope glycoprotein G2 gene was amplified from four specimens of Shandong province by RT-PCR, and the product recombined into the PMD-18T vector. The clones that carry the G2 gene were identified. After sequencing, the gene sequence was handled with the software DNASTAR, compared with 24 strains worldwide and the phylogenetic tree was drawn.

RESULTSHV G2 gene was amplified by RT-PCR from 4 specimens, named GM04-38.G2, ZB8.G2, JUN5-14.G2, RCH5.G2, respectively. The map of the phylogenetic tree showed that all the 4 strains belonged to SEO-type hantavirus. The analysis of the sequence showed that all the four HV strains had the highest rates of homology with Z37 strain. The sequence homology of SEO-type HV strains was from 82.3% to 99.8%.

CONCLUSIONThe four cloning vectors containing the glycoprotein G2 genes were successfully constructed. Envelope glycoprotein G2 gene of four specimens from Shandong province had high homology rates.

Animals ; China ; Cloning, Molecular ; Hantavirus ; genetics ; Mice ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; Viral Envelope Proteins ; genetics

OBJECTIVETo study the surgical reconstruction of secondary fracture deformities of the nasal-orbital ethmoid area.

METHODSTypical bicoronal and subciliary incisions or the adjacent scar incision were employed to expose the fractured area. The flattened nasal bone was narrowed by curved osteotomy along the medial orbital rims and trimed with a bur. 2-3 pieces of cranial outer table were used to reconstruct the nasal framework, which were fixed to the frontal bone with mini-plates. After the medial orbital wall and orbital floor were exposed, the herniated orbital contents were released and reduced to the orbital cavity. The fractured orbital walls were repaired precisely with autogenous cranial outer table or Medpor. The medial canthal tendons were anchored superior-posteriorly to the lacrimal fossa with transnasal wires.

RESULTSFrom December 1996 to December 2001, 34 cases of severe nasal-orbital ethmoid fracture deformities were repaired with this technique. Of them, 12 cases had combined orbital-zygomatic fracture, 4 cases had fontal sinus fracture, 1 case had Le Fort II and 1 case had Le Fort III fracture. All the patients recovered well and their facial appearance improved greatly.

CONCLUSIONSThe key points for surgical reconstruction of the periorbital post-fracture deformities are narrowing the flattened nasal root by curved osteotomy, the nasal framework reconstruction with autogenous bone graft, the orbital wall repair to correct enophthalmos, and most importantly, the medial canthal tendon reduction and canthal plasty.

Adolescent ; Bone Transplantation ; methods ; Cicatrix ; surgery ; Enophthalmos ; surgery ; Ethmoid Bone ; injuries ; Humans ; Nasal Bone ; injuries ; Nose Deformities, Acquired ; surgery ; Orbit ; surgery ; Orbital Fractures ; surgery ; Osteotomy ; methods ; Paranasal Sinuses ; injuries ; surgery ; Polyethylenes ; Reconstructive Surgical Procedures ; Skull Fractures ; etiology ; surgery ; Tendons ; surgery

OBJECTIVETo investigate ligand-gated cation channels P2X3 receptors changes in rat pulp during experimental tooth movement (ETM), and preliminarily find their possible effect during ETM.

METHODS54 male SD rats (200-250 g) were selected and randomly divided into blank group (5 rats), control group (14 rats) and experimental group (35 rats). The left maxillary first molar was selected as observation object, the pulp tissue biopsies was taken at different time points to carry out immunohistochemical study.

RESULTSPredominant up regulation of P2X3 receptors immunoactivity was found in pulp from 1/6 d to 7 d after experimental tooth movement. It started to significantly increase at 1/6 d, peaked at 3 d, and then decreased continuously until 7 d as compared with the beginning.

CONCLUSIONP2X3 receptors have a rhythm change in rat pulp as a result of ETM, speculated that P2X3 receptors is closely related to the tooth movement injury, but the mechanism of action need further researches.

Animals ; Cations ; Dental Pulp ; Male ; Molar ; Rats ; Rats, Sprague-Dawley ; Receptors, Purinergic P2X3 ; Tooth Movement Techniques

AIMTo detect the expression of VASA in human ejaculated spermatozoa, and to compare the expression of VASA between normozoospermic men and patients with oligozoospermia.

METHODSEjaculated spermatozoa were collected from normozoospermic men and patients with oligozoospermia by masturbation, and subsequently segregated through a discontinuous gradient of Percoll to obtain the spermatozoa. Reverse transcription polymerase chain reaction (RT-PCR), quantitative RT-PCR (QRT-PCR), immunoflurescence and Western blotting were used to detect the expression of VASA in mRNA and protein levels.

RESULTSVASA mRNA was expressed in the ejaculated spermatozoa. QRT-PCR analysis showed that VASA mRNA level was approximately 5-fold higher in normozoospermic men than that in oligozoospermic men. Immunofluorescence and Western blotting analysis showed that VASA protein was located on the cytoplasmic membrane of heads and tails of spermatozoa, and its expression was significantly decreased in oligozoospermic men, which is similar to the result of QRT-PCR.

CONCLUSIONThe expression of VASA mRNA and protein was significantly decreased in the sperm of oligozoospermic men, which suggested the lower expression of the VASA gene might be associated with pathogenesis in some subtypes of male infertility and VASA could be used as a molecular marker for the diagnosis of male infertility.

Biomarkers ; metabolism ; Blotting, Western ; DEAD-box RNA Helicases ; genetics ; metabolism ; Fluorescent Antibody Technique, Indirect ; Gene Expression ; Humans ; Male ; Oligospermia ; genetics ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Spermatozoa ; cytology ; metabolism

OBJECTIVETo explore the effect of three-dimensional structure of dermal matrix on biological behavior of fibroblasts (Fb) in the microcosmic perspective.

METHODSThe three-dimensional structure of dermal tissue was analyzed by plane geometric and trigonometric function. Microdots structure array with cell adhesion effect was designed by computer-assisted design software according to the adhesive and non-adhesive components of dermal tissue. Four sizes (8 microm x 3 microm, space 6 microm; 16 microm x 3 microm, space 6 microm; 16 microm x 5 microm, space 8 microm; 20 microm x 3 microm, space 2 microm) of micropier grid used for cell culture (MPGCC) with cell-adhesive microdots, built up with micro-pattern printing and molecule self-assembly method were used to culture dermal Fb. Fb cultured with cell culture matrix without micropier grid was set up as control. The expression of skeleton protein (alpha-SMA) of Fb, cell viability and cell secretion were detected with immunohistochemistry, fluorescent immunohistochemistry, MTT test and the hydroxyproline content assay.

RESULTSThe three-dimensional structure of dermal tissue could be simulated by MPGCC as shown in arithmetic analysis. Compared with those of control group [(12 +/- 3)% and (0.53 +/- 0.03) microg/mg, (0.35 +/- 0.04)], the expression of alpha-SMA [(49 +/- 3)%, (61 +/- 3)%, (47 +/- 4)%, (51 +/- 3)%] and the content of hydroxyproline [(0.95 +/- 0.04), (0.87 +/- 0.03), (0.81 +/- 0.03), (0.77 +/- 0.03) microg/mg] were increased significantly (P < 0.05), the cell viability of Fb (0.12 +/- 0.03, 0.13 +/- 0.04, 0.14 +/- 0.03, 0.19 +/- 0.03) cultured in MPGCC was decreased significantly (P < 0.05). When the parameters of micropier grid were changed, the expression of alpha-SMA, the cell viability and the content of hydroxyproline of Fb cultured in four sizes of MPGCC were also significantly changed as compared with one another (P < 0.05).

CONCLUSIONSMPGCC may be the basic functional unit of dermal template, or unit of dermal template to call. Different three-dimensional circumstances for dermal tissue can result in different template effect and wound healing condition.

Biocompatible Materials ; Cell Adhesion ; Cell Culture Techniques ; Cells, Cultured ; Computer-Aided Design ; Dermis ; Fibroblasts ; cytology ; Tissue Engineering ; methods