Objective To explore the role of mouse dermis-derived mesenchymal stem cells (mdMSC) on skin repair. Methods mdMSC and human dermal fibroblasts were isolated and identified. Human dermal fibroblasts were cultured alone or eoeultured with mdMSC in Transwell chambers with the density ratio of human dermal fibroblasts to mdMSC being 2/5, 1/1, and 2/1. On day 4 and 8 of culture, the expression levels of hydroxyproline and transforming growth factor-beta (TGF-beta) 1 were measured in the supematant of monoculture and coculture by alkaline hydrolysis and ELISA respectively. Results The level of hydro-xyproline was significantly higher in the supematants of coculture system with a density ratio of 2/5 and 1/1 than that in monoculture supematants of human dermal fibroblasts on day 8 (both P < 0.05). Elevated level of TGF-betal was observed in all coculture supematants on day 8 (all P < 0.01) and in the supernatants of coculture system with a density ratio of 1/1 on day 4 (P < 0.05). There was no significant correlation between the expression level of TGF-betal and hydroxyproline in the coculture supernatants (r = 0.108, P ＞ 0.05). Conclusion In vitro coculture with mdMSC can increase the production of hydroxyproline and TGF-betal by fibroblasts, which may be a mechanism underlying the facilitation of skin repair by mdMSC.

OBJECTIVETo explore the feasibility of exact logistic regression, used as a complemental method for the maximum liklihood estimation, and to analyse with data small sample, unbalanced structure and highly stratal nature under the situations of questionable results or inexistence of the maximum likelihood estimation.

METHODSData from 37 postoperative breast cancer cases were analyzed in 1997 by exact logistic regression under SAS system.

RESULTSData calculated by SAS software showed that Quasi-complete separation of data points was detected but the results of maximum likelihood estimation did not exist, SAS outputs conflicted the results of the last maximum likelihood iteration (likelihood Chi-square and score Chi-square have similar P, less than 0.05, but the Wald chi-square had a larger P, more than 0.05). Under conditional exact parameter estimation, it appeared that: (1) the joint effect of conditional score statistics was 21.12 with P = 0.000 3; (2) for individual parameters, the effect conditional score statistics of histological classification (grades) was 5.80 with P = 0.020 8; axillary node metastasis (diversion) was 5.74 with P = 0.019 5; tumor size (size) was 0.79, with P = 0.647 6. The effects of tumor histological classification and axillary node metastasis were statistically significant on breast cancer tumour.

CONCLUSIONExact logistic regression seemed to be a very useful method in analyzing data from small sample when the maximum likelihood estimation was either with no effect or did not exist.

Adult ; Breast Neoplasms ; epidemiology ; pathology ; China ; epidemiology ; Female ; Humans ; Logistic Models ; Middle Aged ; Monte Carlo Method ; Prognosis ; Software

Objective To observe the effects of 75 gram glucose oral tolerance test (75 g OGTT) and standard mixed meal test (SMMT) on insulin secretion function of the islets of Langerhans and plasma free fatty acid (FFA) in patients with type 2 diabetes mellitus.Methods Seventy-six patients with type 2 diabetes without using insulin and with no obvious complications were recruited for 75 g OGTT following overnight fasting on the first day and SMMT (bread 50 g,egg 50 g and milk 250 ml) on the 7th day.Blood specimens were collected from each patients before the tests and 30 min,60 min,120 min and 180 min after glucose or meal load to measure their levels of plasma glucose,serum insulin,C peptide,FFA and lipids (total cholesterol,triglyceride,high-density and low-density lipoprotein cholesterol).Results No difference in fasting plasma glucose,serum insulin,C peptide,FFA and lipids between 75 g OGTT and SMMT was found.Postprandial plasma glucose 30 min,60 min,120 min and 180 min after 75 g OGTT was significantly higher than that after SMMT,with (15.3?3.5) vs (9.9?3.4) mmol/L,(18.2?4.8) vs (12.8?4.0) mmol/L,(16.3?5.8) vs (12.2?4.9) mmol/L and (10.6?5.4) vs (9.5?4.5) mmol/L (F=28.1,P

OBJECTIVETo investigate the clinical effectiveness of sertraline hydrochloride combined with four-spot caress in the treatment of primary premature ejaculation (PE).

METHODSWe randomly assigned 90 primary PE patients to three groups of equal number. The patients in group A (aged [28.1 ± 5.2] yr and with a disease course of [3.1 ± 1.9] yr) were treated with oral sertraline hydrochloride at 50 mg qd, those in B (aged [27.8 ± 4.1] yr and with a disease course of [3.2 ± 2.0] yr) by four-spot caressing (caressing the tongue, breasts, and vulva prior to intercourse), and those in C (aged [27.1 ± 4.7] yr and with a disease course of [3.1 ± 2.0] yr) by the combination of oral sertraline hydrochloride and four-spot caressing, all for 12 weeks. Before and after 4, 8, and 12 weeks of treatment, we obtained the intravaginal ejaculatory latency time (IELT) and Chinese Index of Sexual Function for Premature Ejaculation-5 (CIPE-5) scores and compared them among the three groups of patients.

RESULTSThe IELT was dramatically prolonged in groups A, B, and C after 4 weeks ([1.08 ± 0.29], [0.93 ± 0.28] and [1.21 ± 0.27] min), 8 weeks ([1.43 ± 0.30], [1.20 ± 0.33] and [1.72 ± 0.42] min) and 12 weeks of treatment ([2.12 ± 0.63], [1.90 ± 0.65] and [2.67 ± 0.82] min) as compared with the baseline ([0.63 ?0.14] , [0.60 ?0.14] and [0.62 ?0.11] min) (P < 0.05), even longer in group C than in A and B (P < 0.05). The CIPE-5 scores were markedly improved in groups A, B and C after 4 weeks ([15.17 ± 1.74], [14.57 ± 1.94] and [15.60 ± 1.63] min), 8 weeks ([17.13 ± 1.63], [16.37 ± 1.97] and [18.00 ± 1.05] min) and 12 weeks of intervention ([18.93 ± 1.57], [18.53 ± 1.67] and [20.00 ± 1.46] min ) as compared with the baseline ([12.57 ± 2.05], [13.20 ± 2.51] and [13.07 ± 2.01] min) (P < 0.05), even higher in group C than in A and B (P < 0.05).

CONCLUSIONSertraline hydrochloride combined with four-spot caressing, with its definite efficacy and rare adverse reactions, deserves wide clinical application in the treatment of primary PE.

Adult ; Coitus ; Ejaculation ; Female ; Humans ; Male ; Premature Ejaculation ; drug therapy ; Sertraline ; therapeutic use ; Young Adult

Objective To dig the correlation between genes and four TCM constitutions (phlegm-dampness, blood stasis, allergy, and dampness-heat); To analyze microscopic mechanism of constitution formed and constitution-disease correlation; To provide biological basis for TCM constitution classification and constitution-disease correlation theory. Methods Literature about TCM constitutions and gene expressions in CNKI and Wanfang Database were retrieved by computers from the establishment of database to 31, December 2016. Differentially expressed genes of four TCM constitutions were collected, and DAVID database was used for enrichment analysis, including gene tissue enrichment analysis, disease analysis, GO analysis, and pathway analysis. The results of DAVID database analysis were downloaded and analyzed. Results Four TCM constitutions had differences in gene level, and each constitution had the specificity in significantly enriched gene function, activity, pathways, related diseases, and expression locations. Different patterns of gene expression led to the constitution differences in disease susceptibility and tendency. Conclusion The essence of TCM constitution formation is the change of gene expression patterns, and the pathological changes are also caused by the change.

In order to obtain the gene P12X3C of Foot-and-Mouth Disease Virus (FMDV) that includes full length P1, 2A, 3C and a part of 2B, the site mutation strategy was used. After being digested by Kpn I and Xba I respectively, the gene P12X3C was cloned into the pcDNA3.1 (+) expression vector. The recombinant plasmid was checked by restriction enzyme analysis and nucleic acid sequencing, and then named pcDNA3.1/P12X3C. Further, BHK-21 cells was transfected with pcDNA3.1/P12X3C by using lipoid. The proteins of Foot-and-Mouth Disease Virus, which were expressed in BHK-21 cells, were confirmed by sandwich-ELISA and fluoroscopy. The result shows the gene P12X3C is cloned into eukaryotic expression plasmid, and the recombinant eukaryotic expression plasmid pcDNA3.1/P12X3C could express proteins of Foot-and-Mouth Disease Virus in BHK-21 cells, which have immunocompetence. This study demonstrates that delivery of a recombinant eukaryotic expression plasmid containing P12X3C coding regions results in the assembly of FMDV capsid structures, which will offer experimental base to DNA vaccine of FMDV.
Animals ; Cell Line ; Cricetinae ; Enzyme-Linked Immunosorbent Assay ; Fluoroscopy ; Foot-and-Mouth Disease Virus ; genetics ; Genetic Vectors ; genetics ; Models, Genetic ; Plasmids ; genetics ; Viral Proteins ; genetics ; metabolism

OBJECTIVETo explore a way of the gene therapy for acute spinal cord injury (ASCI) by vivo transfection of exogenous gene into spinal cord tissue.

METHODSTwenty-four rats of SD were divided into experiment group and control group (each group had 12 rats). After anaesthesia by abdominal cavity, lamina of thoracic vertebra of all rats were cut-open in prone position. Complex of plasmid and report gene-Lac Z, and plasmid without report gene-Lac Z were respectively injected into cavum subdural of SD rats of experiment group and control group by cation liposome (DOTAP) encapsulation. The rats were killed at the 2nd week after operation, spinal cord tissue of injected segments were detected by reverse transcription-polymerase chain raction (RT-PCR) and immunohistochemistry.

RESULTSIn experiment group, positive staining of beta-galactosidase can be clearly observed in neuron and glia cell of rat's spinal cord by immunohistochemistry detection. Lac Z mRNA in same area was also detected by RT-PCR. But, in control group, no above-mentioned positive results were found.

CONCLUSIONEffective transfection of exogenous gene in vivo into spinal cord is a new hot spot for treatment of SCI. Thus certain nerve growth factor imput partly area of spinal cord injury can promote central nerve regrowth and avoid early secondary injury.

Acute Disease ; Animals ; Genetic Therapy ; Lac Operon ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Spinal Cord ; metabolism ; Spinal Cord Injuries ; therapy ; Transfection ; beta-Galactosidase ; analysis

OBJECTIVETo analyze the differential proteomics in human umbilical cord mesenchymal stem cells (MSC) induced by chemical hypoxia-mimetic agent cobalt chloride (CoCl(2)) by two-dimensional gel electrophoresis (2-DE) and mass-spectrometry.

METHODS2-DE was performed to separate proteins from treated and untreated human umbilical cord MSC with CoCl(2). 2-DE images were analyzed by ImageMaster 2D Platinum software 6.0. The differential expressed proteins was identified by MALDI-TOF-MS. The differential proteins were classified based on their functions.

RESULTS2-DE reference patterns of CoCl(2) treated human umbilical cord MSC were established. A total of twenty-six differential proteins were identified, of them eleven proteins were up-regulated and fifteen down-regulated. Their biological functions involved in carbohydrate metabolism, protein metabolism and modification, lipid metabolism, coenzyme and prosthetic group metabolism, cell cycle, immunity and defense, cell structure and motility, signal transduction, protein targeting and localization, neuronal activities, muscle contraction, etc. Peroxiredoxin1 (Prdx) was down-regulated, whereas alpha-enolase (ENO1) and vesicle amine transport protein 1 homolog (VAT1) up-regulated.

CONCLUSIONThe effects of hypoxia on human umbilical cord MSC were participated by multiple proteins and involved in multiple functional pathways.

Cobalt ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; drug effects ; metabolism ; Proteome ; analysis ; Proteomics ; Umbilical Cord ; cytology ; drug effects

OBJECTIVETo observe the results of posterior osteotomy and correction in the surgical treatment of old tuberculous kyphosis.

METHODSFrom June 2004 to December 2008, 31 cases of old tuberculous kyphosis with posterior osteotomy and correction technique were treated. There were 12 cases of male and 19 cases of female. The average age was 33.4 years. Pedicle subtraction osteotomy or vertebral column resection were applied in surgery. The kyphosis angle, lumbar lordosis angle and sagittal balance condition of the spine were measured before and after surgery, as well as follow-up. The Frankel grading system for neurological function of lower extremities, the Oswestry disability index (ODI) for life quality, and patient satisfactory index (PSI) for satisfaction of surgery were applied before surgery and at follow-up.

RESULTSThe average kyphosis angle was 94° ± 27°, the average lumbar lordosis angle was 71° ± 20°, and the average sagittal C(7) plumb line was (-15 ± 44) mm away from the balance region before surgery. The average kyphosis angle decreased to 26° ± 11° in one week after surgery, with an improvement rate of 71.4%. The average follow-up time was 22.5 months. The average kyphosis angle was 28° ± 12° at the final follow-up, with an improvement rate of 70.0%. The average lumbar lordosis angle was 46° ± 11°, with an improvement rate of 35.1%. The postoperative kyphosis angle and lumbar lordosis angle were significantly different with that of pre-operation (for kyphosis angle: t = 16.3, P < 0.05; for lumbar lordosis angle: t = 8.1, P < 0.05). The average sagittal C(7) plumb line was (-4 ± 22) mm away from the balance region at the final follow-up, with an improvement rate of 73.4%. The Frankel grading were E in 13 cases, D in 13 cases, and C in 5 cases before surgery, and were E in 20 cases, D in 8 cases, and C in 3 cases at the final follow-up. The average ODI was 13 ± 12 before surgery, and was 7 ± 8 at the final follow-up, with an improvement rate of 45.2%. The PSI results showed a satisfied rate of 90.3%.

CONCLUSIONGood results can be achieved by applying proper posterior osteotomy and correction technique according to the severity of old tuberculous kyphosis.

Adolescent ; Adult ; Female ; Follow-Up Studies ; Humans ; Kyphosis ; etiology ; surgery ; Male ; Middle Aged ; Osteotomy ; methods ; Spinal Fusion ; methods ; Treatment Outcome ; Tuberculosis, Spinal ; complications ; Young Adult

OBJECTIVETo design a new surgical correction and fixation technique for the treatment of severe angular kyphosis, observe the feasibility, safety and effectiveness of the technique.

METHODSFrom May 2004 to February 2007, 16 cases with severe kyphosis (average 90.8 degrees, range 50 degrees-130 degrees) were treated with segmental resection osteotomy, section distraction, dual axial rotation correction and instrumentation fusion technique. The patients were inspected by local and total spine anteroposterior and lateral radiography pre-and postoperatively. The kyphotic Cobb angle was measured and 7 cases combined scoliosis Cobb angle was also measured. The Frankel Grading for neurological function, Oswestry Disability Index (ODI) and Patients Satisfactory Index (PSI) were evaluated preoperatively, postoperatively and at follow-up. The back pain relief was also observed.

RESULTSThe average surgical duration was 6.9 hours. The average blood loss was 4000 ml. The complications include 1 shifting of artificial vertebrae, 3 nerve root injury, 3 dural tear and 1 transitory dysfunction of lower extremity. All of these complications were relieved greatly after feasible treatment. The average follow-up time was 25 months. The average kyphotic angle was 90.8 degrees preoperatively, which was improved to 26.9 degrees immediately after surgery, and got an average correction rate of 72.5%. At follow-up, the average kyphotic angle was 28.9 degrees, and correction rate was 70.1%. The 7 cases who combined with scoliosis had an average Cobb angle of 35.9 degrees preoperatively, which decreased to 4.4 degrees immediately after surgery, and the correction rate was 87.2%. The correction rate was kept until follow-up (78.6%). Some patients got an improved neurological function. The Frankel Grading were E in 5 cases, D in 5 cases, C in 5 cases, and B in 1 case preoperatively. There were 10 cases of E grade, 3 cases of D grade, and 3 cases of C grade at follow-up. Except 3 cases who had no symptoms before surgery, the average ODI was 18.9 preoperatively, and 10.8 postoperatively. The average improvement of ODI was 52.7%. The PSI result showed a satisfied rate of 93.8%. The back pain of 3 cases were totally relieved after surgery.

CONCLUSIONSSegmental resection osteotomy with dual axial rotation correction and fusion technique is an effective way to treat severe angular kyphosis. It is a safe technique and has high correction rate. The long-term results is acceptable.

Adolescent ; Adult ; Aged ; Bone Transplantation ; methods ; Child ; Feasibility Studies ; Female ; Follow-Up Studies ; Humans ; Kyphosis ; surgery ; Male ; Middle Aged ; Osteotomy ; methods ; Spinal Fusion ; methods ; Treatment Outcome