Objective To explore the efficient way to purify and to enrich umbilical cord mesenchymal stem cells which have abilities to differentiate into chondrocytes. Methods Umbilical cord was acquired from new born infants and chopped into 1 mm2 cubics, then they were pretreated with colleganase and trypsin so as to get the mixed cell suspension. CD45(-),CD90(+) cells were acquired from the mixed cell suspension via FCM sorting and were cultured in the medium as umbilical cord mesenchymal stem PO cells. The PO cells were detached and passaged by multiplex collagenase NB4 and the positive rate of CD90 and CD45 of the P1-P3 cells were observeed via FCM ; the P3 cells were induced to differentiated into chondrocytes. Results FCM results showed that the positive rate of CD90 on P1, P2, F3 was (80.86 ± 7.85)%,(95.86± 3.28)%,(97.15± 1.43)%, while the positive rate of CD45 on P1, P2, P3 cells was (2.53 ±0.28)% and (0.97 ± 0.48)% and (0.05 ± 0.01)% respectively; P3 cells had differenciated into chondrocytes. Conclusion FCM presorting and Multiplex collagenase detachment procedures can swiftly acquire highly purified human umbilical cord mesenchymal stem cells which have the ability to differentiate into chondroeytes.

Objective To investigate the brain protective effect of isoflurane preconditioning on the rat liver against ischemia-reperfusion through determining the content of S-100β protein in peripheral blood in combination with mitochondrial ultrastructure in rat brain.Methods A total of 45 SD rats were randomly divided into three groups,sham group (group S):the only separation of the hepatoduodenal ligament,but did not block the hepatic portal blood supply; ischemia-reperfusion group (group I/R):liver ischemia 60min,reperfusion 120 min; isoflurane preconditioning group (group ISO):60 min before liver I/R,ISO pretreatment for 30 min,elution in the air after 30 min; 24 h after recirculation the forebrain tissues were rapidly removed.The changes of mitochondrial ultrastructure were observed by electron microscopy.The content of S-100β protein in serum was measured before ischemia and reperfusion 120 min through the application of Elisa kit.Results Marked swelling of mitochondria with disrupted cristae and damaged matrix were observed in group I/R,while relative intact mitochondria were seen in sham and ISO groups.The content of S-100β protein in serum was significantly higher in I/R group [(1.52 ±0.26) μg/ml] than in sham [(0.31 ±0.05)μg/ml] and ISO [(0.79 ± 0.21) μg/ml] groups (P ＜0.05).Conclusions The liver ischemia-reperfusion may injure the brain of the rat and isoflurane preconditioning can protect the rat brain from injury.

Objective To study protective immunity effects of co-immunization with P30 DNA vaccine and protein vaccine. Methods Forty-eight 5-6 weeks old BALB/c female mice were divided into four groups (A,B,C,D), 12 mice of each group. In group A (control group) each mouse was immunized with 100 ?g pcDNA3.1 plasmid DNA by intramuscular (i.m.) for three times at week 0,2 and 4; in group B (P30 protein group) each mouse was immunized (i.m.) with 50 ?g rP30+50 ?g CFA for three times at week 0, 2 and 4; in group C (pcDNA3.1-P30 group) each mouse was immunized with 100 ?g pcDNA3.1-P30 plasmid DNA (i.m.) for three times at week 0, 2 and 4; in group D (P30 DNA+rP30 co-immunization group) each mouse was immunized with 100 ?g pcDNA3.1-P30 plasmid DNA (i.m.) for two times at week 0, 2 and immunized by subcutaneous with 50 ?g rP30+50 ?g CFA at week 4. Each mouse was infected with 100 tachyzoites of Toxoplasma gondii RH strain four weeks later after last immunization. The anti-P30 antibodies were detected with ELISA before the challenge. Results The P30 DNA vaccine was successfully constructed. High titers of anti-P30 antibodies were induced in each mouse immunized with DNA vaccine. The protective trial proved that there was no significant difference between control group and experimental group though the survival time of mouse from experimental group had been prolonged. Conclusion The P30 DNA vaccine could induced high titers of anti- P30 antibodies in immunized mice, and it may be a potential DNA vaccine candidate.

Cell Line, Tumor ; Cystadenocarcinoma, Serous ; pathology ; Female ; Humans ; Ovarian Neoplasms ; pathology

Recently, microRNAs (miRNAs) have been shown to be involved in multiple biological pathways that can influence tumor progression and metastasis and they can serve as prognostic biomarkers in many cancers. The present study examined the prognostic significance of miR-215 in cervical cancer. The paraffin-embedded paired cervical scrape samples and tumor tissue samples from 302 patients with stage II cervical cancer were detected for the expression of miR-215 by using qRT-PCR. A miR-215-based classifier was established by using the Cox regression model. The prognostic and predictive accuracy of this classifier was determined in both the internal testing group of 138 patients, and the external independent group of 280 patients. Moreover, cervical cancer HeLa cells overexpressing miR-215 (HeLa-miR-215) were constructed and subcutaneously injected into the nude mice to examine the effect of miR-215 on tumor growth and metastasis in vivo. The results showed that the expression level of miR-215 was significantly higher in cervical cancer tissues than in paired normal tissues (P<0.0001). When patients were classified into high- and low-risk cancer progression groups according to miR-215 level, the 5-year disease-free survival in high- and low-risk groups were 43% (95% CI: 32.1-51.6) and 67% (95% CI: 48.6-77.3) (hazard ratio [HR] 2.02, 95% CI: 1.16-3.52; P=0.013) respectively. Moreover, the expression level of miR-215 was negatively associated with survival rate in patients at TNM stage T3 (HR: 3.317; 95% CI: 1.18-5.14, P=0.017) and TNM stage T4 (HR: 3.48; 95% CI: 1.49-4.45, P=0.008). Tumor volume in nude mice injected with HeLa-miR-215 cells was significantly larger than that in mice injected with control HeLa cells. It was concluded that the expression level of miR-215 is associated with cervical tumor progression and worse survival rate, suggesting that it may serve as a potential prognostic marker to identify patients at higher risk of recurrence.

Objective:To investigate the predictive value of different glycemic variability indexes within 6 hours on the short-term prognosis of septic shock patients.Methods:A retrospective study was conducted. The 133 patients with septic shock admitted to intensive care unit (ICU) of Nanjing Hospital Affiliated to Nanjing Medical University from December 2014 to December 2019 were enrolled. Patients with septic shock admitted to ICU died during hospitalization were enrolled in the death group and others in the survival group. General data of the patients including gender, age, underlying disease, site of infection, duration of mechanical ventilation, length of ICU stay, whether to use continuous renal replacement therapy (CRRT) and acute physiology and chronic health evaluationⅡ (APACHEⅡ) scores within 24 hours were collected. The blood glucose (GLUadm), mean arterial pressure (MAP), serum creatinine (SCr) and procalcitonin (PCT) were recorded at ICU admission. The patients admitted to ICU received bundle therapy within 6 hours and blood glucose was observed every 2 hours. The blood glucose difference (GLUdif), average blood glucose (GLUave), blood glucose standard deviation (GLUsd) and blood glucose variation coefficient (GLUcv) within 6 hours were calculated. Multivariate Logistic regression analysis was used to analyze the prognostic factors of short-term prognosis of patients with septic shock, and receiver operating characteristic (ROC) curve was plotted to evaluate the diagnostic efficacy of glycemic parameters for short-term prognosis of septic shock patients.Results:A total of 133 patients with septic shock were admitted to ICU, among them 87 patients survived and 46 patients died during the ICU hospitalization. Compared with the survival group, the SCr at ICU admission and APACHEⅡ score within 24 hours were significantly higher in the death group [SCr (μmol/L): 208.5 (143.0, 286.5) vs. 172.0 (91.0, 234.0), APACHEⅡ score: 30.28±6.67 vs. 24.03±5.90, both P < 0.05], the length of ICU stay was shorter [days: 4.00 (2.00, 10.25) vs. 9.00 (4.00, 13.00), P < 0.01]. However, there were no significant differences in the baseline data of gender, age, underlying disease, infection site, CRRT ratio, MAP or PCT at ICU admission between the two groups. Compared with the survival group, the GLUsd and GLUcv within 6 hours in the death group were higher [GLUsd (mmol/L): 2.33 (1.95, 3.14) vs. 2.02 (1.66, 2.52), GLUcv: (31.00±7.06)% vs. (23.31±10.51)%, both P < 0.05]. There were no statistically significant differences in the levels of GLUadm, GLUdif or GLUave within 6 hours between the two groups. Multivariate Logistic regression analysis showed that APACHEⅡ score within 24 hours and GLUsd and GLUcv within 6 hours were independent risk factors of the short-term prognosis of septic shock patients [APACHEⅡ score: odds ratio ( OR) = 1.173, 95% confidence interval (95% CI) was 1.095-1.256, P = 0.000; GLUsd: OR = 1.465, 95% CI was 1.038-2.067, P = 0.030; GLUcv: OR = 1.089, 95% CI was 1.043-1.138, P = 0.000]. ROC curve analysis showed that GLUsd and GLUcv within 6 hours both had certain predictive value for the short-term prognosis of septic shock patients, the area under ROC curve (AUC) of GLUcv within 6 hours was higher than that of APACHEⅡ score (0.765 vs. 0.753), and AUC of GLUsd within 6 hours was close to APACHEⅡ score (0.629 vs. 0.753); and the diagnostic value of GLUsd combined with GLUcv within 6 hours was higher than the two respectively (AUC: 0.809 vs. 0.629, 0.765), the sensitivity was 97.8%, and the specificity was 66.7%. Conclusion:GLUsd combined with GLUcv within 6 hours can be used to estimate the short-term prognosis of septic shock patients.

Objective:To investigate the predictive value of serum thyroid hormone in the short-term prognosis of patients with septic shock..Methods:The 133 patients with septic shock admitted to Intensive Care Unit (ICU) of Nanjing Hospital of Nanjing Medical University were enrolled. Data of patient’s general clinical information, acute physiology and chronic health evaluationⅡ (APACHE Ⅱ), serum triiodothyronine ( T3 ), thyroxine (T4), free triiodothyronine (FT3) , free thyroxine (FT4) and thyroid stimulating hormone(TSH) in 24 h were collected, and the ratio of T3 to FT3 (T3/FT3) was calculated. The patients were divided into the survival group and death group according to the prognosis during ICU hospitalization. Univariate and multivariate analyses were used to analyze the prognostic factors of patients with septic shock. The receiver operating characteristic (ROC) curve and Kaplan-Meier analysis were plotted to evaluate the diagnostic efficacy of serum thyroid hormone for the short-term prognosis of patients with septic shock.Results:Compared to the survival group, FT3, T3 and T3/FT3 were significantly lower in the death group [1.73 (1.54, 2.52) vs. 1.32 (0.94, 1.54) pmol/L, 0.70 (0.56, 0.79) vs. 0.33 (0.25, 0.43) nmol/L, 318.18(299.44, 448.05) vs. 250.00 (192.31, 313.92), respectively; all P < 0.05 ]. The levels of serum T3 and FT3 were significantly correlated with the APACHEⅡ score in patients with septic shock (FT3: r = -0.25, P= 0.004; T3: r = - 0.24, P= 0.006). Binary Logistic regression analysis showed that FT3, T3 and T3/FT3 were independent risk factors of the short-term prognosis of patients with septic shock [FT3: OR = 6.533, 95% CI: 0.687 - 62.157, P = 0.012; T3: OR = 0.529, 95% CI: 0.372 - 0.975, P= 0.000; T3/FT3: OR = 1.719, 95% CI: 1.007 - 1.931, P= 0.002]. ROC curve analysis showed that FT3, T3 and T3/FT3 all had certain predictive value for the short-term prognosis of patients with septic shock, and the diagnostic value of T3 was the largest [AUC = 0.874, 95% CI: 0.794-0.954, P= 0.000]. The Kaplan-Meier curve showed a significantly higher survival rate in patients with T3 greater than 0.535 nmol/L than patients with T3 less than 0.535 nmol/L. Conclusions:The serum levels of thyroid hormone T3 and FT3 are closely related to the severity of septic shock. T3 is an effective ICU mortality during hospitalization prognostic indicator for patients with septic shock, which is better than FT3 and T3/FT3.

The essential medium of B115 composed of 1% tryptone, 0.25% yeast extract and 0.5% sodium chloride was determined by using an orthogonal design. The orthogonal design was also employed in testing the optimum additions. It was composed of 0.1%(NH_(4))_(2)SO_(4),1.4%K_(2)HPO_(4), 0.6% KH_(2)PO_(4) and 0.1% (Na_(3)C_(6)H_(5)O_(7)). The yield of B115 cultured in optimum medium was compared with the one in essential medium. Statistic analysis showed that the growth of B115 was most significantly improved by adding K~(+)、NH~+_(4) and (Na_(3)C_(6)H_(5)O_(7)) to essential medium. The antibacterial effect of Bacillus subtilis strain B115 on pathogenic Aeromonas was studied. The results showed different antibacterial effects of B115 on different aeromonads. There were obvious antibacterial effects on BSK-10 and CL990920, while no effect on the growth of TL970424.

Objective To investigate the early clinical outcome of peripheral cutting balloon(PCB) in the management of peripheral vessel stenosis.Methods Thirteen patients with peripheral limb vessel stenosis, in which 4 stenoses in hemodialysis access and 9 at lower limb arteries,underwent angioplasty by PCB.For multiple stenosis in the same vessel,the distal one should be expanded firstly.The balloon pressure was controlled in the range of 8 atm to 10 atm(1 atm=10.108 kPa).All the patient were given continuous anticoagulant therapy after the procedure.Results All the procedure were carried out successfully on the 13 patients,and no serious complications occured.The symptoms did not recur in all patients after the procedure.The 5 months' follow-up angiography proved that no restenosis occurred in one patient with previous stenosis at the hemodialysis access.Conclusion The angioplasty with PCB was a safe and reliable procedure in management of the peripheral limb vessel stenosis.The early outcome is satisfying.

Objective: To study the infectious state of 3 kinds of viruses (CVB, CMV and EBV) in respiratory tract infection and its significance. Methods: Among 290 patients studied, there were 158 boys and 132 girls, aged from 6 months to 12 years. The antigen of coxsackievirus B (CVB-Ag) and antibody of coxsackievirus B (CVB-IgM), antibody of cytomegalovirus (CMV-IgM) and antibody of EB virus (EBV- IgM) were detected by ELISA method. Results: (1) The positive incidence of upper respiratory tract infection, bronchitis, pneumonia was higher than that of noninfectious disease group (P<0.01). The positive incidence of CVB was higher than that of CMV and EBV(P＜0.01). (2)There were cross viruses positive, especially in pneumonia group (P<0.05), usually CVB cross other virus infection. (3) The course of pneumonia was long in single CVB (P<0.01) and longer in cross infection of viruses (P<0.01). Conclusion: CVB infection is the most common one in the 3 viruses. Attention should be paid to multiple viruses infections in pneumonia children.