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.

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 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 .

Objective To investigate the clinical value of laparoscopic resection of gastric stromal tumors.Methods Thirty-two cases received this new type of operation.The tumors ranged in size from 1.5 to 5.5cm with a mean diameter of 2.6 cm.The operative methods included the full laparoscopic resection of(gastric) tumor and the hand-assisted laparoscopic resection of gastric tumor.Results All cases were(successfully) resected and no complications were observed.The mean operative time was 75min.The mean(intraoperative) blood loss was 50ml.Postoperative pain was slight.Postoperative flatus and feces were passed at a mean of 34 hours,and average postoperational hospital stay was 7.5days.Postoperative pathologic(examination) confirmed that 25 cases were benign GIST and 7 cases were of low-grade malignancy.No(recurrences) were observed at follow up of 8 to 30 months.Conclusions Laparoscopic resection of gastric(stromal) tumor is a technically simple,safe and effective procedure that could be widely used.

Objective:To investigate the prognostic factors and pathological characteristics of mixed subtype thyroid cancer (MSTC) .Methods:Data of 41 cases of MSTC, which were confirmed by postoperative pathology, among from 24, 912 cases of thyroid cancer admitted in Mar. 2005 to Aug. 2020 in the First Medical Center of Chinese People’s Liberation Army General Hospital, were retrospectively analyzed. 37 cases underwent surgical treatment, while 4 cases only underwent puncture to confirm the pathology due to physical conditions, and no surgical treatment was performed. The tumor size, number of lesions, capsule invasion, AJCC 8th TNM staging, surgical methods, radiotherapy and chemotherapy were collected. The MSTC patients in the group were followed up to obtain the postoperative situation. SPSS 25.0 and R studio statistical software was used for data processing, and Cox single factor and multivariate regression were used to analyze independent risk factors.Results:In the 41 cases, there were 9 cases of papillary carcinoma (PTC) mixed with follicular carcinoma (FTC) , and 8 cases of mixed medullary and follicular carcinoma (MMFTC) . There were 15 cases of poorly differentiated thyroid cancer (PDTC) , 4 cases of squamous cell carcinoma of thyroid (SCCT) , and 5 cases of undifferentiated thyroid carcinoma (ATC) . The median follow-up time was 18 months, and 11 patients died during the follow-up, with a mortality rate of 26.8%. Average onset age was (51.41+15.69) years. 4 cases had postoperative recurrence during the follow-up, including 2 cases of local recurrence, and 2 cases of distant metastasis. Single factor results showed that age, degree of tumor differentiation, surgical method, radiotherapy and chemotherapy were the risk factors affecting the prognosis of patients with MSTC ( P<0.05) . Multivariate analysis showed that age at diagnosis ( P=0.007) and surgical procedure ( P=0.017) were independent risk factors for prognosis in patients with MSTC. Conclusion:Middle-aged and elderly women are at high risk for MSTC, and the degree of tumor differentiation is proportional to survival. Due to the multi-type and pleomorphic pathological findings, a reasonable treatment plan has good effects on prognosis of MSTC.

Objective:To explore the influencing factors of false-positive serological reaction of syphilis.Methods:A total of 166 patients with false-positive serological reaction of syphilis (false-positive group), 145 patients diagnosed with early syphilis without treatment (positive control group) and 124 persons undergoing entry physical examination (negative control group) were included from January 2017 to February 2020 in Beijing Tongren Hospital, Capital Medical University. The gender, age and underlying diseases of the three groups were compared. Logistic regression was used to analyze the influencing factors of false-positive serological reaction of syphilis. The efficacies of chemiluminescence immunoassay (CLIA) and toluidine red unheated serum test (TRUST) were compared. Paired t test or chi-square test was used for statistical analysis. Results:In the false-positive group (166 cases), the age of 117 cases were more than 50 years old and 49 cases <50 years old. There were significant differences in age ((53.1±13.8) vs (24.7±2.8), t=22.56, P<0.01), autoimmune disease (36.7%(61/166) vs 6.5%(8/124), χ2=35.93, P<0.01), hepatitis (9.6%(16/166) vs 3.2%(4/124), χ2=4.92, P=0.026) and tumor (6.6%(11/166) vs 0.8%(1/124), χ2=4.68, P=0.030) between the false-positive group and the negative control group. There were significant differences in gender (there were 91(54.8%) males and 75(45.2%) females in the false-positive group, and 103(71.0%) males and 42(29.0%) females in the positive control group, χ2=8.67, P=0.003), age ((53.1±13.8) vs (34.4±12.9), t=20.13, P<0.01) and autoimmune disease (36.7%(61/166) vs 6.9%(10/145), χ2=39.14, P<0.01) between the false-positive group and the positive control group. Multivariate logistic regression analysis showed that gender (odds ratio ( OR)=2.692, 95% confidence interval ( CI) 1.504-4.816, P=0.001), age ≥50 years old ( OR=30.512, 95% CI 15.959-58.335, P<0.01), autoimmune disease ( OR=2.677, 95% CI 1.258-5.695, P=0.011) and hepatitis ( OR=4.408, 95% CI 1.799-10.799, P=0.001) were the influencing factors of false-positive serological reaction of syphilis. In the false-positive group, the positive rate of TRUST was 84.9% (141/166), which was higher than that of CLIA (23.5%(39/166)). The difference was statistically significant ( χ2=126.25, P<0.01). CLIA was 1.0-10.0 cut off index (COI) in 36 patients, and >10.0 COI in three patients.The difference was statistically significant ( χ2=52.51, P<0.01). The titers were ≤1∶4 in 139 patients and≥1∶8 in two patients with TRUST positive.The difference was statistically significant ( χ2=262.35, P<0.01). The sensitivity and specificity of CLIA were 95.2% and 96.0%, respectively, and those of TRUST were 77.2% and 91.1%, respectively. Conclusions:The influencing factors of false-positive serological reaction of syphilis include patients age ≥50 years and with autoimmune disease or hepatitis. The false-positive rate of TRUST is significantly higher than CLIA.