Objective: To clone the full-length cDNA encoding squalene epoxidase 1 (SQE1), a key enzyme of triterpenes biosynthesis, from Pseudostellaria heterophylla and to perform functional analysis. Methods: With the total RNA as template, the full-length cDNA of SQE1 in P. heterophylla was cloned via RT-PCR and rapid amplification of cDNA ends (RACE) techniques. The bioinformatics of the cloned SQE1 gene was performed. The target gene was transfered into tobacco by Agrobacterium-mediated transformation. Results: The full-length cDNA (2 038 bp) of SQE1 gene was obtained with an open reading frame of 1 554 bp, encoding 517 amino acid polypeptides, which had higher homology with the known SQEs in other medicinal species. The calculated relative molecular mass was 5.67 × 104, the isoelectric point was 8.8. The deduced protein sequence exhibited FAD-binding domains and four transmembrane regions. The content of total triterpenes was increased in transgenic tobacco plants. Conclusion: This is the first report that the full-length cDNA encoding SQE1 from P. heterophylla is cloned. The ectopic expression of SQE1 could promote to increase the content of total triterpenes in transgenic plant. This work provides a foundation for exploring the biosynthetic pathway of triterpenes in P.heterophylla and their applications in bioengineering.

To study the effect of sphingosine-1-phosphate (S1P) on the cardiomyogenic differentiation of human umbilical cord mesenchymal stem cells (UC-MSCs) and human adipose-derived mesenchymal stem cells (AD-MSCs), we seeded the cells in the culture plates and used cardiomyocyte culture medium (CMCM) combining with different concentration of S1P to induce UC-MSCs and AD-MSCs in vitro for 7, 14 and 28 days. Cardiomyogenic differentiations were identified through immunofluorescence staining, and the results were observed with fluorescence microscopy and confocal microscopy. The effects of S1P and CMCM on cell activity were evaluated by the methyl thiazolyl tetrazolium assay. The functional characteristic similar to cardiomyocytes was evaluated through detecting calcium transient. Our results showed that cardiomyogenic differentiation of UC-MSCs or AD-MSCs were enhanced with S1P concentration increasing, but cell activities declined. Results showed that the suitable differentiation time was 14 days, and the optimal concentration of S1P was 0.5 micromol/L. When working together with CMCM, S1P could promote the differentiation of UC-MSCs or AD-MSCs into functional cardiomyocytes, giving rise to specific electrophysiological properties (the calcium transient). Taken together, our results suggested that S1P could promote the differentiation of UC-MSCs or AD-MSCs into functional cardiomyocytes when being cultured in CMCM.
Adipose Tissue ; cytology ; metabolism ; Cell Differentiation ; drug effects ; Cells, Cultured ; Culture Media ; Humans ; Lysophospholipids ; pharmacology ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Myocytes, Cardiac ; cytology ; Sphingosine ; analogs & derivatives ; pharmacology ; Umbilical Cord ; cytology

Objective Although the correlation between high risk human papilloma virus (hrHPV) infection and cervical cancer ( CC ) or cervical intraepithelial neoplasia ( CIN ) is well known , vaginal cancer ( VC ) or vaginal intraepithelial neoplasia ( VAIN) also caused by hrHPV has not received enough attention .This article aims to explore the clinical characteristics of VC or VAIN after operations of CC or CIN in order to provide evidence for the treatment of these diseases . Methods The clinical charac-teristics and treatment of 15 cases with VC or VAIN after operations of CC or CIN were reviewed from Jan 2010 to May 2013 in our hos-pital. Results The mean age was (53.6 ±10.82) years, ranged from 39 to 73 years.The duration from the first operation to devel-oped VAIN or VC was (25.07 ±18.31) months, ranged from 1 to 60 months.There are 4 cases developed VC, 4 cases VAINⅢand 2 cases VINⅡfrom 10 CC patients;and 3 cases developed VC , 2 cases VAINⅢfrom 5 CINⅢpatients.hrHPV test were positive in all 15 patients.Treatment in these series were performed including total vaginectomy in 8 patients (3 VC, 4 VAINⅢ and 1 VAINⅡpatients), pelvic lymphonectomy in 1;upper vaginectomy in 2 patients (1 VC, 1 VAINⅢ), radiation or chemo-radiation therapy in 3 (3 VC), interferon muscle injection combined with topical application of estrogen and acyclovir gel in 2 (1 VC, 1 VAINⅡ). Conclusion Careful follow-up after CC or CIN operations are very important because continued hrHPV infection may result VC and VAIN lesions.Vaginectomy may be the best therapy .Interferon muscle injection combined with topical application of estrogen and acyclovir gel are also alternatively therapy , especially for hard to operate patients . Radiation therapy seems to be not very adaptable for VAIN patients .

OBJECTIVE: To study the percentages of polymorphonuclear leukocytes (PMN), mononuclear cells (MNC) and fibroblastic cells (FBC) in different post-traumatic intervals after skeletal muscle mechanical injury in rats. METHODS: The rat model of skeletal muscle mechanical injury was established. The rats were divided into injured groups (6 h, 12 h, 1 d, 3 d, 7 d, 10 d and 14 d after injury) and control group. The percentages of PMN, MNC and FBC in different post-traumatic intervals after skeletal muscle mechanical injury were assessed with HE staining and image analysis. RESULTS: At post-injury 6-12h, the percentages of PMN and MNC infiltration appeared in injured sites and that of PMN reached peak. At 1 d, the percentage of MNC infiltration appeared and reached peak, while that of PMN decreased. At 3-7 d, the percentage of FBC gradually increased, while that of PMN and MNC decreased. At 10-14d, the percentage of FBC reached peak. CONCLUSION The percentages of PMN, MNC and FBC in injured zones showed time-dependent changes, which might be used as reference index for determination of age of skeletal muscle injury.
Animals ; Fibroblasts ; Muscle, Skeletal/injuries* ; Neutrophils ; Rats ; Time Factors

Purpose To investigate the clinicopathlogical characteristics, diagnosis and differential diagnosis of ovarian small cell car-cinoma hypercalcemic type ( OSCCHT) associated with acute renal dysfunction. Methods A case of OSCCHT associated with acute renal dysfunction was reported. The clinical and pathologic data, treatment and pathological examinations were analyzed and the related literatures were reviewed. Results A 29-year-olds women was presented to hospital with inappetence and significant weight loss for 2 months. The laboratory examination showed abnormal renal function, and pelvic cavity mass, possibly coming from adnexa of the uterus was seen by radiography. Serum levels showed significantly increased serum calcium with acute renal dysfunction. An emergency opera-tion was performed just after renal function partially recovered by 2 times hemodialysis. During surgery, right ovary tumor with a size of 12 cm × 10 cm × 10 cm was inspected. Microscopically, the tumor cells were arranged in a diffuse solid pattern, mutiple nodules were separated by fibrous tissue. some small folliculars and pseudoglandular cavities with acidophilia secretion within nodules could been ob-served. Tumor cells were medium to large with eosinophilic cytoplasm, round or oval vesicular nucleus, increased karyoplasmic ratio and pathologic mitosis. Immunohistochemistry revealed that the tumor cells expressed EMA, CKpan, C-erbB-2 ( +) , CA125 ( focal+) and Ki-67 proliferation index was about 60%, while ER, PR, Syn, CgA, PTH, Inhibin, CD99, AFP, PLAP, CD30 and CD20 were not expressed. Conclusion Ovarian small cell carcinoma hypercalcemic type with acute renal dysfunction is a very rare with a rapidly progressive and highly malignant tumor. The final diagnosis mainly lies on clinical information ( hypercalcaemia) , morphology and immunohistochemistry, combined with electron microscopy and molecular biological detection when necessary.

OBJECTIVETo inquire into the relationship between lipoprotein lipase (LPL) gene D9N, N291S and S447X polymorphisms and the development of cardiovascular diseases in children with obesity.

METHODSThe polymerase chain reaction (PCR) and restriction fragment length polymorphism (RLFP) techniques were used to detect three common mutations of LPL gene exon D9N, N291S and S447X in 157 obese children and 175 normal controls. Plasma lipid and lipoprotein levels between children with different genotypes were compared.

RESULTSThe D9N and N291S gene mutations were not detected in either the obese or the control groups. There were no significant differences in the frequency of S447X gene mutation between the two groups. There were no significant differences in the levels of plasma lipid and lipoprotein between children with S447 and X447 genotypes.

CONCLUSIONSD9N and N291S gene mutations may not be risk factors associated with cardiovascular diseases in children with obesity. S447X gene mutation might not play an important role in the development of cardiovascular diseases in childhood.

Adolescent ; Cardiovascular Diseases ; etiology ; genetics ; Child ; Female ; Humans ; Lipoprotein Lipase ; genetics ; Male ; Mutation ; Obesity ; genetics ; Risk Factors

OBJECTIVE: To research the relation between the time-dependent appearances of myotibroblasts during the repair of contused skeletal muscle in rat and wound age determination. METHODS: A total of 35 SD male rats were divided into the control and six injured groups according to wound age as follows: 12 h, 1 d, 5 d, 7 d, 10 d and 14 d after injury. The appearances of myofibroblasts were detected by HE staining, immunohistochemistry and confocal laser scanning microscopy. Masson's trichrome staining was utilized to examine collagen accumulation in the contused areas. RESULTS: Immunohistochemical staining showed that α-SMA+ myofibroblasts were initially observed at 5 d post-injury. The average ratio of myofibroblasts was highest at 14 d post-injury, with all samples, ratios more than 50%. In the other five groups, the average of α-SMA positive ratios were less than 50%. The collagen stained areas in the contused zones, concomitant with myofibroblast appearance, were increasingly augmented along with advances of posttraumatic interval. CONCLUSION The immunohistochemical detection of myofibroblasts can be applied to wound age determination. The myofibroblasts might be involved in collagen deposition during the repair of contused skeletal muscle in rat.
Animals ; Collagen/metabolism* ; Contusions/metabolism* ; Immunohistochemistry ; Male ; Microscopy, Confocal ; Muscle, Skeletal/metabolism* ; Myofibroblasts/metabolism* ; Rats ; Time Factors ; Wound Healing

OBJECTIVETo establish a three-dimension finite element model of mandible with two kinds of dental implant and to study the stress of implant-bone interface.

METHODSMeasuring the data of the components of the dental implant and using spiral CT image reconstruction technique to scan the cross section of the mandible. Three-dimension finite element analysis software Unigraphics and MSC. Marc/Mentat were used to build the conjunction model and bone model of two implant systems. Loading 200 N axially and 100 N 30 degrees obliquely on the models respectively, the stress distribution patterns of the bone interface of two implant systems were analyzed.

RESULTSThe stress distribution on the bone interface of two implant systems was similar. The peak stress of oblique loading was higher than that of axial loading. The peak stress district of the bone was concentrated on the stricture of the implant cervix, which was more obviously displayed on the Replace Select implant. The peak stresses on the bone interface of Replace Select implant were higher than that of Replace implant in all loadings.

CONCLUSIONTo Replace Select especially, oblique force should be avoided in clinical practice in case of the bone absorption.

Computer Simulation ; Dental Implants ; Finite Element Analysis ; Humans ; Mandible ; Stress, Mechanical

UNLABELLED: OBJECTIVE To investigate the time-dependent expression of c-jun during the healing of incised wound in mice skin. METHODS: The expression of c-jun in different stages after the incised wound were detected by immunohistochemistry and Western blot. RESULTS: There was a low level expression of c-jun in normal mice skin. Expression of c-jun was mainly detected in neutrophils from 3 h to 12h after injury. The c-jun positive cells were almost mononuclear cells (MNCs) and fibroblasts between 1 d and 5 d after injury. The c-jun positive cells were mostly fibroblasts between 7 d and 14 d after injury. The ratio of the c-jun positive cells increased in the wound specimens from 3 h to 12 h, peaked at 12 h, declined partially from 1 d to 5 d, and reached the peak secondly at 7 d, then decreased from 10 d to 14 d. The expression of c-jun was observed throughout the wound healing stages by Western blot with two peaks occurring at 12 h and 7 d after injury. CONCLUSION The c-jun may play a potential role in inducing apoptosis of neutrophils, MNCs and fibroblasts during skin wound healing, and it may be used as the marker for wound age determination.
Animals ; Apoptosis ; Blotting, Western ; Disease Models, Animal ; Female ; Fibroblasts/metabolism* ; Immunohistochemistry ; Male ; Mice ; Neutrophils/metabolism* ; Proto-Oncogene Proteins c-jun/metabolism* ; Random Allocation ; Skin/metabolism* ; Time Factors ; Wound Healing ; Wounds and Injuries/metabolism*

The myofibroblasts have dual characteristics of smooth muscle cells and fibroblasts. In repairing tissular wound, myofibroblasts are involved in fibrogenesis and remodeling the extracellular matrix of the fibrotic cascades reaction. The review describes the morphological characteristics and biological behaviors of myofibroblasts and the application of skin wound age determination, which may provide reference for research in forensic medicine.
Actins/metabolism* ; Animals ; Cell Differentiation ; Cells, Cultured ; Extracellular Matrix/metabolism* ; Fibroblasts/physiology* ; Forensic Pathology/methods* ; Humans ; Muscle, Smooth/physiology* ; Myofibroblasts/physiology* ; Skin/injuries* ; Time Factors ; Wound Healing ; Wounds and Injuries/pathology*