OBJECTIVETo explore the role of the basic fibroblast growth factor (bFGF) in the directional differentiation of bone marrow mesenchymal stem cells (BMSCs) into Leydig cells.

METHODSAfter purification and identification, we inoculated the third-generation BMSCs of SD rats onto a six-orifice board and then randomly divided them into groups A (normal saline control), B (human chorionic gonadotropin [hCG] + platelet-derived growth factor [PDGF] induction), C (hCG + PDGF + 5.0 ng/ml bFGF induction), D (hCG + PDGF + 10.0 ng/ml bFGF induction), and E (hCG + PDGF + 20.0 ng/ml bFGF induction). On the 7th, 14th and 21st day of induction, we observed the morphological changes of the cells and measured the level of testosterone (T) and expression of 3 beta hydroxy steroid dehydrogenase (3β-HSD) in the supernatant by immunofluorescence staining.

RESULTSAfter induction, the BMSCs of groups B, C, D, and E exhibited microscopic features of enlarged size, inter-connection, long-shuttle or irregular shape, adherent growth, and large round nuclei, all characteristic of Leydig cells. With the prolonging of time and enhanced concentration of bFGF, gradual increases were observed in the T level and the count of 3β-HSD-positive BMSCs in the four induction groups, with statistically significant differences between group B and groups C, D, and E (P < 0.05), as well as between group C and groups D and E (P < 0.05), but not between D and E (P > 0.05).

CONCLUSIONThe bFGF has an obvious promoting effect in the in vitro induced differentiation of rat BMSCs into Leydig cells.

Animals ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Cell Differentiation ; Cells, Cultured ; Chorionic Gonadotropin ; metabolism ; Fibroblast Growth Factor 2 ; pharmacology ; Humans ; Leydig Cells ; cytology ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Testosterone ; metabolism

Objective To study the efficacy of transcutaneous prostaglandin E 1( PGE 1) cream in relaxing the vascular spasm in rabbits, to find an ideal vascular dilation drug. Methods Sixty rabbits were randomly divided into six groups:experiment group 1(using 0.1% transcutaneous PGE 1 cream),experiment group 2(using 0.2% transcutaneous PGE 1 cream),experiment group 3(using 0.4% transcutaneous PGE 1 cream), experiment group 4( using 0.8% transcutaneous PGE 1 cream),PGE 1 control group (without using transcutaneous PGE 1 cream, but using PGE 1 cream only),control group(withous using PGE 1). 0.1% adrenlin was injected to the root of the rabbite ear to induce a model of vascular spasm. After a typical vascular spasm appeared,transcutaneou PGE 1 cream or PGE 1 cream was used on the skin of the ear. The vascular diameter and blood folw rate of the ear were determined before adrenalin injection, 10min after vascular spasm and 10,15,30,60,90 and 120 min after the drugs were used. Results The vascular diameters and blood flow rate were recovered to the levels of before adrenalin injection in experimental group 3, 4 after topical used of transcutaneous PGE 1 cream, and those parameters were significamly increased compared with other 4 groups(P0.05). Conclusions Tramscutameou PGE 1 cream can effectively relax the vascular spasm caused be adrenalin injection.The effect of vascular dilation is related to the dosage of the drug. Transcutaneous PGE 1 cream as a topical vascular dilation drug has advantages snch as wide indications,taking effect quick and enduring, and less side effect. So it might be a new simple, safe and effective drug for the treatment of vascular spasm.

This study was to appraise safety and feasibility of laparoscopic approach and investigate the clinical effects of laparoscopic tension-free repair of esophageal hiatal hernia using mesh. From August 2006 to July 2009, 24 patients with esophageal hiatal hernia underwent laparoscopic repair. Twenty-three patients received laparoscopic tension-free repair using mesh, at the same time, Toupet or Dor partial fundoplication was performed. One patient was converted to open surgery. The average operating time was 90 min (70-210 min) and the blood loss was between 10-110 mL. There was no death. The mean postoperative hospital stay was 5 days (3-30 days). During a follow-up period of 12-20 months (mean 15 months), there was no recurrence of the hernia, and no complication with use of mesh. The present study suggested that laparoscopic approach was secure and minimally invasive operation for esophageal hiatal hernia and the use of mesh could reduce recurrence rate.

Objective To investigate the expression and significance of ABCG2 and ALDH1 in malignant transformation of ovarian endometriosis.Methods The protein expressions of ABCG2 and ALDH1 were detected with immunohistochemistry in 38 cases of malignant transformation of ovarian endometriosis(EMs malignant transformation group),35 cases of endometriosis(EMs group)and 30 cases of normal endometrium(control group).Results The positive rates of ABCG2 and ALDH1 expressions in EMs malignant transformation group were significantly higher than those in EMs group and control group(P<0.05).The expressions of ABCG2 and ALDH1 were significantly associated with the serum CA125 level,differentiation and clinical stage of malignant transformation of ovarian endometriosis (P<0.05),but were not related to the patient′s age,dysmenorrhea history and pathology type(P>0.05).There was no correlation between the expression of ABCG2 and ALDH1 in malignant transformation of ovarian endometriosis(P>0.05).Conclusion The expressions of ABCG2 and ALDH1 might be involved in malignant transformation of ovarian endometriosis.

Objective To clone and express surface antigen SAG4 gene of Toxoplasma gondii, and analyze its immunoreactivity. Methods Specific primers were designed based on the reported SAG4 gene of T. gondii RH strain (GenBank Accession No: AF340224.1). Using genomic DNA from T. gondii as templates, SAG4 gene was amplified by PCR. The PCR product was cloned into pMD19-T vector and identified by digestion with restriction enzyme and PCR. Then the target fragment was subcloned into pET28a(+) vector, transformed into E. coli BL21 and followed by expression of the protein induced by IPTG. The protein was identified by Western blotting. Results The target gene was amplified with the length of 537 bp. Sequence analysis showed that the predicted amino acid sequence was identical with that of SAG4 as a membrane protein in T. gondii. After induced by IPTG, the recombinant SAG4 protein existed in an inclusion body form. The recombinant SAG4 (Mr 18 740) was recognized by serum of infected mice. Conclusion SAG4 has been expressed and shows certain immuno-response activity.

Objective To prepare and identify the monoclonal antibody against asymmetric dimethylarginine (ADMA) and to do further research on clinical application .Methods The short immune peptide antigen was synthetized by solid phase technology . Splenocytes from the Balb /c mice immunized by synthetized antigen were fused with myeloma cells SP 2/0 for producing hybrido-ma .Hybridoma cell line secreting antibodies against ADMA was sifted by indirect ELISA and limiting dilution assay .The specificity of monoclonal antibodies against human ADMA were evaluated with Western blot and ELISA .Results Two hybridoma cell lines stably secreting monoclonal antibodies against ADMA were developed and named 22-1-1and 38-1-6 respectively .By applying West-ern blot and ELISA ,the results indicated that all monoclonal antibodies raised could specifically react with ADMA .Conclusion The success in the production of ADMA monoclonal antibody establishes foundation for application in the practice of clinical labora-to ry .

This article analyzes and summaries the problems and difficulties existing in the first-aid medicine's clinical teaching.Combined with his own teaching experience,the author has made some exploration on the teaching time,the way of teaching and inspection form,which has got a good teaching effect.It provids the reference and development for the clinical teaching of first-aid medicine.

Objective To detect the levels of CD+4 CDHi25 CDLo127 regulatory T (Treg) cells and NKG2D-expressing NK cells in peripheral blood of lung cancer patients and investigate their relationship,and explore their roles in the immune escape of lung cancer and clinical significance.Methods The levels of CD+4 CDHi25 CDLo127 Treg cells,NK cells and NKG2D-expressing NK cells were measured in peripheral blood of 70 lung cancer patients and 50 healthy controls.Results Compared with control group,the levels of CD+4 CDHi25 CDLo127 Treg cells enormously increased [(8.4±4.1) ％,(6.7±1.7) ％] (t =3.09,P ＜ 0.05),while the value of NKG2D obviously decreased [(83.3±4.9) ％,(87.4±2.9) ％] (t =3.16,P ＜ 0.05); And the group of NK cells is not different [(15.6±8.3) ％,(17.2±4.2) ％] (t =-1.33,P ＞ 0.05); NKG2D-expressing NK cells had a perfect negative correlation with CD+4 CDLo25 CDLo127 Treg cells (r =-0.302,P ＜ 0.05).Conclusion Accumulation of Treg cells in serum may lead to the down-modulation of NKG2D-expressing NK which participates the tumor immune escape.They may be potential indicators evaluating immune functions and prognosis of lung cancers.

Objective To investigate early changes of angiopoietin-2(Ang-2)in multiple trauma patients and assess its clinical significance.Methods Forty-five multiple trauma patients aged 2060 years admitted to the hospital within one hour after injury were randomly divided into three groups according to injury severity score(ISS).Blood specimens were obtained immediately upon arrival in the emergency department and plasma samples were assayed for comparing changes of Ang-2,TNF-a and IL-6.Meanwhile,plasma level of Ang-2 was measured and analyzed under different oxygenation index,shock index and base deficit.Results Plasma level of Ang-2 was positively correlated with ISS(P ＜ 0.01)and was concordant with the plasma levels of TNF-a and IL-6(P＜0.01).Furthermore,plasma level of Ang-2 was elevated upon increase of shock index or decrease of oxygenation index(P ＜ 0.01).Plasma level of Ang-2 was elevated with the increase of base deficit(P ＜ 0.01).Conclusions High level of Ang-2 is a marker of endothelial activation and dysfunction early after trauma.Ang-2 is related tightly with the injury severity,inflammation factors,systemic oxygenation and tissue hypoperfusion and may have a tight relation with pathophysiological development and clinical outcome after trauma.

Objective To investigate the thermal stability of solid potassium iodate and potassium iodate as additive in the sodium chloride,vitamin E,vitamin C and yellow prussiate.Methods HCR-2 type Differential Thermal Analyzer was used to carried out the differential thermal analysis of the potassium iodate and the potassium iodate in the sodium chloride,vitamin E,vitamin C and the yellow prussiate,and differential thermal curves were obtained and analyzed.Results The decomposition temperature of solid potassium iodate was 525℃ ; when mixed with sodium chloride,potassium iodate was stable below 300 ℃ ; vitamin C was unstable at 170-200 ℃ and underwent chemical changes; iodate and vitamin C underwent oxidation-reduction reaction at 145 to 160 ℃;potassium iodate with vitamin E at 300 ℃ was stable; yellow prussiate at 300 ℃ was stable; iodized salt was stable at cooking temperature below 300 ℃.Conclusions The potassium iodate has good stability below 525 ℃,however,potassium iodate iodized salt in the cooking process is easy to react with vitamin C in vegetables causing iodine losses,so iodized salt should be added just before the dish is done.