Medical science is the science which closely combines scientific knowledge with humanistic spirit. Becausemodern science and technology have been more and more applied in medical research and practice, the medical sciencehas been developed rapidly. But medical science calls us to concern more about medical humanistic spirit, so how wecombine medical scientific knowledge with humanistic spirit is a very important topic in our medical university education.[

Objective To study the aberrant DNA methylation of H19 and MEST in infertile male sperm.Methods Routine semen analysis and morphological analysis were performed on 15 control individuals and 33 oligoastheno-teratozoospermias.Sperms were purified by density gradient centrifugation and DNA was extracted and treated with bisulfite.After PCR,the purified PCR products were cloned into pMD (R)18-T vector and proceed to chemical transformation,restriction enzyme reaction.For each sample,positive clones were selected for sequencing analysis and DNA methylation analysis.Results Five control individuals had high degree of H19 and methylation rate was 100.0％,while the methylation rate of 93.0％ in 10 oligoastheno-teratozoospermias was significantly lower.Significant difference was found in methylation rate between the control individuals and oligoastheno-teratozoospermias (P ＜ 0.01).As to MEST,10 control individuals had a low degree of MEST and the methylation rate was 1.6％,while the methylation rate in 23 oligoastheno-teratozoospermias was 4.8％.No significant difference was found in the methylation rate between control individuals and oligoastheno-teratozoospermias (P ＞ 0.05).Conclusion Male infertility is associated with aberrant methylation of H19 differentially methylated region,and it may be a biomarker of human spermatogenesis defect.

Objective To study the differentially expressed microRNA (miRNA) between pancreatic ductal adenocarcinoma (PDAC) and para-cancerous tissues,and determine related target genes.Methods Nine fresh PDAC tumor tissues and 3 adjacent normal pancreatic tissues were collected,then Agilent miRNAmicroarray with 713 miRNA loci was used to identify the differentially expressed miRNA.Real-time PCR method was applied to verify the up-regulated miRNA.TargetScan 5.1 and miRandaV5 software were used to analyze the related target genes.Results miRNA microarray identified 11 PDAC related miRNAs,among them,the expressions of miR-194*,miR-192*,miR-602,miR-194 were up-regulated,while the expressions of miR-139-3p,miR-513a-5p,miR-630,miR-30c-1 *,miR-887,miR-508-5p,miR-516a-5p were down-regulated.The expressions of miR-192,miR-194 and their homolog were verified in 31 PDAC tumor tissues.After software analysis,it was found that target genes of miR-192 were ZEB2,CXCL-2,EEF1A1,ERCC3,and target genes of miR-192 * were DCC,SMAD4,FAS,and target genes of miR-194 included DACHI,IGSF11,PTPN2,RBBP4,while target genes of miR-194 * included CD40LG,CIDEB,FHL1.Conclusions Eleven differentially expressed miRNAs are present in PDC,and they may be involved in the occurrence and development of PDC.

BACKGROUND:Cryotherapy plays a positive role in the treatment of delayed-onset muscle soreness caused by high intense exercise. OBJECTIVE: To investigate the effects of different crypotherapy programs on the levels of interleukin-6 and prostaglandin 2 in long distance race-walkers after 15-day training, and to determine a rational treatment program for delayed-onset muscle soreness. METHODS:Sixteen male race-walkers in Liaoning Province were randomly divided into cryotherapy and cryo/heat therapy groups, and received 10-minute cryotherapy and 2.5-minute cryo/heat therapy (2.5-mintue cryotherapy and 2.5-minute heat therapy alternately for 10 minutes), respectively, after 15-day training. The serum levels of interleukin-6 and prostaglandin 2 were detected at six different time points to compare the efficacy between two methods. RESULTS AND CONCLUSION: Compared with the cryo/heat therapy group, the serum levels of interleukin-6 and prostaglandin 2 in the cryotherapy group were significantly decreased. That is to say, cryotherapy is more available for alleviating delayed-onset muscle soreness after intensive eccentric training or in intensive seasons.

Objective To study the influence of site-directed deglycosylation of the HIV-1 envelope (Env) on its immunogenicity and assembly of functional pseudovirus. Methods Site-directed deglycosylation were performed using cycling mutagenesis and selection of mutants with DpnⅠ. Single-cycle infection assay was employed to analyze the effect of the mutations on the ability of functional pseudovirus assembly. The influence of deglycosylations on the immunodeficiency of Env was evaluated using pseudovirusbased neutralization assay and ELISPOT assay. Results Mutant N197Q induced higher neutralization activities for both pseudoviruses, but lower Env-specific T-cell response. And N197Q rendered the Env to lose the ability of functional pseudovirus assembly. Mutant G2 induced higher neutralization activities for pseudovirus 74-2 but lower for pseudovirus Wt, and had almost no influence on Env-specific T-cell response and functional pseudovirus forming. Conclusion The site-directed deglycosylation of the HIV-1 Env affects the pseudovirus forming and its immunogenicity.

Objective To study the in vivo expression and biodistribution of Ad5-Fluc (Adenovirus carrying firefly luciferase genes) in mice.Methods The recombinant Ad5-Fluc virus was constructed and infected to BALB/c or nude mice through three different routes.The protein expression level,tissue distribution and the characteristics of infection were analyzed by in vivo bioluminescence imaging technology.Results Compared to other two routes,the BALB/c mice infected through muscular route had the longest expression cycle (over 60 days) and the highest expression level,while the virus was transferred into the liver and spleen after infection.The nude mice had a significantly extended expression cycle than BALB/c mice.Moreover,the characteristic of liver tropism was eliminated after Ad5 F35 infection in mice,while maintained similar expression efficiency.Conclusion Due to the highest expression efficiency,the muscular route would be the optimal route for Ad5 vector based vaccination.In addition,Ad5F35 virus could become an ideal alternative vaccine vector for eliminating the liver tropism.

Objective To investigate the expression of HSPB1 in pancreatic cancer and its relationship with clinicopathological characterization.Methods Pathological specimens from 47 cases of pancreatic cancers,13 cases of para-cancerous normal pancreatic tissues and 3 cases of chronic pancreatitis tissues were selected,and tissue microarray construction instrument was used to prepare tissue microarray,then the expression of HSPB1 was determined by immunohistochemistry SP method.The scores of the proportion of positive cells and staining intensity were multiplied to make the judgment.Results The expression of HSPB1 in malignant,para-cancerous and chronic pancreatitis tissues was 80.9％ (38/47),12.5％ (2/16),respectively; and the difference between the two groups was statistically significant (X2 =24.058,P =0.000).The expression of HSPB1 in pancreatic cancer tissue was not significantly related to sex,age,location,differentiation degree and nerve infiltration of tumor ( P ＞ 0.05 ).Conclusions The expression of HSPB1 is related to development and progression of pancreatic cancer,and may be an early molecular event.

AIM: To study the chemical constituents from the leaves of Phyllanthus emblica L. METHODS: The constituents were extracted by percolation with 95% ethanol.Then the extract was separated by systemic solvent separation methods.The ethyl acetate portion from the leaves were separated and purified by silica gel column chromatography and polyamide column chromatography with gradient elution,liquid preparation and recrystal methods.The structures of crystals were identified by physiochemical properties,spectrum analysis and literatures ontrast. RESULTS: Five compounds were isolated and identified.They were ?-sitosterol(Ⅰ);?-carotene(Ⅱ);kaemferol(Ⅲ);quercetin(Ⅳ);avicularin(Ⅴ). CONCLUSION: Chemical compound Ⅴ is isolated from this plant for the first time.

Objective:To evaluate the value of combined measurement of urinary insulin-like growth factor-binding protein 7 (IGFBP7) and urinary metalloproteinase inhibitor-2 (TIMP-2) in the early diagnosis and prognosis of cardiac surgery-associated acute kidney injury (CSA-AKI).Methods:From March 2018 to June 2018, cardiac surgery patients admitted to the cardiac macrovascular surgery department of the First Affiliated Hospital of Nanjing Medical University were prospectively included, and the blood creatinine was monitored to observe the presence of acute kidney injury (AKI). The prognostic information of the patients was collected, including in-hospital dialysis, in-hospital death, complete recovery of kidney function at discharge, death in one year after surgery, and progression to chronic kidney disease. The levels of urine IGFBP7 and TIMP-2 at 6 h, 24 h and 48 h after cardiac surgery were detected by enzyme linked immunosorbent assay (ELISA), and the urine creatinine (Cr) was also measured. Moreover, receiver operating characteristic curves (ROC) were plotted and the areas under the curves ( AUC) were calculated to evaluate the predictive value and prognostic value of urinary [TIMP-2]·[IGFBP7] (T*I for short) and urine T*I/urine Cr 2 in CSA-AKI. Results:A total of 74 patients with age of (58.43±10.91) years old and 47 males, were enrolled in this study, of which 24 cases (32.4%) had AKI and 10 cases (13.5%) had stage 2-3 AKI. Compared with the non-AKI group, the AKI group had significantly higher levels of urine T*I levels at 6 h and 24 h (both P<0.05). The AUC of T*I at 24 h predicting for AKI was 0.71(95% CI 0.59-0.81, P=0.001, cutoff value 0.020, sensitivity 79.2%, specificity 56.0%), while the AUC for stage 2-3 AKI was 0.85 (95% CI 0.75-0.92, P<0.001, cutoff value 0.083, sensitivity 70.0%, specificity 90.6%). Urinary T*I normalized for urinary creatinine excretion did not show better predictive value. In addition, of T*I at 24 h predicting for poor hospitalization outcome, renal recovery, and one year postoperative death, the AUC was 0.82(95% CI 0.71-0.90, P=0.001), 0.80(95% CI 0.66-0.86, P<0.001), and 0.81(95% CI 0.70-0.89, P=0.047), respectively. Conclusion:The combined detection of TIMP-2 and IGFBP7 in urine is expected to be a biomarker for early diagnosis of CSA-AKI and has certain clinical value in predicting the prognosis of CSA-AKI.

The ELISPOT assay is increasingly used for assessing cellular immune responses in clinical trials of HIV-1 or cancer vaccines. However, to date, data from clinical trials do not consistently show that immune responses are correlated with clinical endpoints. This is due in part to the lack of assay standardization and validation across laboratories and therefore, a quality control panel is required to establish competency and comparability amongst different laboratories. In this study peripheral blood mononuclear cells (PBMCs) from healthy individuals were screened and frozen in liquid nitrogen. The recovery and viability of the PBMCs and the frequencies of interferon (IFN)-γ-secreting cells after CEF peptide pool stimulation were detected after various intervals in seven different laboratories. The recovery and viability did not differ significantly after different intervals. Although the frequencies of IFN (interferon)-γ-secreting cells among thawed PBMCs (peripheral blood mononuclear cells) fluctuated after CEF peptide pool stimulation at different intervals, they were not significantly decreased compared with those among fresh PBMCs. However, the viabilities, recoveries and frequencies of IFN-γ-secreting cells differed significantly among the seven laboratories. Our results indicate that cryopreserved PBMCs could be used as a quality control panel for ELISPOT. However, the procedures for ELISPOT need to be standardized amongst different laboratories.