Objective To investigate the current status and changing patterns of hemodialysis in Wenzhou areas of China from 1999 to 2006. Methods Data of blood purification centres of 18 hospitals in Wenzhou areas from 1999 to 2006 were collected. The incidence, prevalence, mortality, the etiology and the relevant factors such as age, gender, dialysis ages, outcomes and causes of death in end stage renal disease(ESRD) patients on hemodialysis(HD) were analyzed retrospectively. Results The incident number and the number of patients increased annually while the mortality remained steady. The male patients outnumbered the female every year, but the male/female ratio was decreasing. The percentage of both young and oldly patients was increasing. The first cause was chronic glomerulonephritis, although the constituent ratio of glomerulonephritis decreased year by year. The constituent ratio of diabetic nephropathy and hypertensive nephropathy increased. The constituent ratio of dialysis age 1-2 years group decreased, ≤1 year, 2-3 years, 3-4 years groups were relatively steady, and 4-5 years, 5-10 years, 10 years groups increased. The number of patients receiving renal transplantation and transferring to peritoneal dialysis increased annually. The leading cause of death was cardiovascular incidence (19.9%), followed by cerebrovascular disorder (10.8%), systemic failure (10.8%), hemorrhagic diseases(4.7%) and infectious diseases(4.3%). The constituent ratio of cardiovascular incidence and cerebrovascular disorder were relatively steady. The constituent ratio of hemorrhagic diseases and systemic failure showed great fluctuation. The constituent ratio of infectious diseases and malnutrition was decreasing. Conclusions In Wenzhou area from 1999 to 2006, the patients number has been increasing annually. The onset age of HD is 30-70 years old, and proves a younger tendency and older tendency. The leading cause is chronic glomeralonephritis. The constituent ratio of diabetic nephropathy and hypertensive nephropathy rises year by year. The long-term survival rate of HD patients is improved. The leading cause of death is cardiovascular accident.

Objective To improve the gene targeting efficiency with C57BL/6 embryonic stem ( ES) cells.Meth-ods Three different genetically modified C57BL/6 ES cell lines, named TLX3, Ai3K and SL, were microinjected into ICR, B6( Cg)-Tyrc-2J and BALB/c mouse blastocysts, respectively.The efficiency was statistically evaluated according to three aspects:blastocyst collection, chimera production and germline transmission.Results None of the three ES cell lines was germline transmitted with B6(Cg)-Tyrc-2J mice as blastocyst donors, while it was achieved with both BALB/c and ICR mouse blastocysts.Compared in the aspect of blastocysts collection, ICR mouse was much better than BALB/c mouse (P<0.05), and the chimera production efficiency of ICR mouse was comparable to that of BALB/c mouse (P =0.115). As to the germline transmission efficiency, that of BALB/c mice is significantly higher than that of the ICR mice ( P<0.01).Conclusions The germline transmission efficiency of BALB/c mouse is highest among these three mouse strains. However, it has the disadvantages of blastocyst collection, developmental delay and zona pellucida fragility, compared with ICR mouse.Therefore, ICR mouse is also a good candidate as blastocyst donor for embryonic stem cell microinjection.

Objective To establish a simple , fast and economic total DNA extraction method to serve the rapid identification of model mouse genotype in large number of mice .Methods Three methods, i.e.phenol extraction, isopropyl alcohol precipitation and mouse ear boiling methods were used to extract the total DNA from ten C 57-rasmodel mice.The purity and yield of DNA obtained by the three methods were compared , and polymerase chain reaction ( PCR) assay was used to compare the efficacy of the three extraction methods .Results Among the three methods , phenol extraction was the best and isopropyl alcohol precipitation was the poorest in DNA yield .In terms of DNA purity , the phenol extraction was the best and the mouse ear boiling method was the poorest .All the three methods could be used to extract the total DNA from mice serving as template of PCR reaction for the mouse genotype identification .The time consumption of three methods are 12.5 hr ,13 hr and 0.18 hr.Mouse ear boiling method was significantly lower than that of the other two methods ( P <0.01 ) ,.The obtained total DNA can be stored at conventional -20℃for 7 days and 30 days later still can be used as a template for PCR reaction .Conclusions Among the three methods studied , the mouse ear boiling method is simple and with the lowest cost , so it is feasible for total DNA extraction in scaled genotyping experiments .

Objective To quantitate coronary tortuosity and provide favorable conditions for understanding the hemodynamic effects caused by coronary tortuosity.Methods We obtained all images from 72 patients who received coronary computed tomo-graphy scanning.After image post-processing,we extracted the medial axis of three main coronary vessel and analysed its coordi-nates on three dimensions.Then we calculated the tortuosity coefficient of coronary artery.Results Tortuosity coefficient was 6.66±7.54 in anterior descending,13.43±12.85 in left circumflex,and 1 7.61 ±7.67 in right coronary artery.We had proved its validity by the changes in morphology with simulated shapes.Conclusion We proposed a new method for quantitating coronary tor-tuosity,by computed tomography coronary imaging.The measurement results would not be affected by projection plane,vessel length or other artificial factors.

Objective To obtain the basic growth parameters of a self-established F1 hybrid CB6F1 C57-ras transgenic mouse model, and to provide basic information for commercialization of this mouse model. Methods F1 hybrid mice (CB6F1) were produced by crossing C57-ras heterozygous transgenic (c-Ha-ras+/-) male mice and wild-type BALB/cJ female mice.The average litter size, weaning rate, sex ratio, growth performance and C57-ras transgenic positive rate were recorded and analyzed.Results The average litter size was eight, weaning rate was 90%, and sex ratio was approximately in accordance with prediction.The average body weight of newborn mice was 1.73 ±0.05 g.The average body weight of 10-week old c-Ha-ras transgenic female and male mice in CB6F1 background was 24.38 ±1.74 g and 29.42 ±1.72g, respectively, which had a significant difference (P<0.01).The c-Ha-ras transgenic positive rate was 46.9%. which was in accordance with genetic rules.Conclusions The F1 hybrid mice (CB6F1) produced by crossing C57-ras heterozygous transgenic ( c-Ha-ras +/-) male mice and wild-type BALB/cJ female mice show normal growth performance and development characteristics, and it can be used for large-scale commercial supply.

Objective:To investigate the characteristics of serum cytokine IL-4 and IFN- γ levels in patients with chronic insomnia with mild cognitive impairment (MCI), and to further explore the relationship between cognitive function and IL-4 and IFN-γ in patients with chronic insomnia.Methods:Sixty-two patients with chronic insomnia were divided into MCI group( n=30) and non-MCI group( n=32) according to the scores of Montreal cognitive assessment(MoCA), mini-mental state examination(MMSE) score and chief complaint of cognitive decline. Pittsburgh sleep quality index(PSQI), Hamilton depression scale(HAMD 24) and Hamilton anxiety scale 14 item(HAMA 14) were evaluated. Serum IL-4 and IFN-γ were detected by flow fluorescence, correlation analysis and regression analysis were carried out. Results:The levels of IL-4 and IFN-γ in MCI group were significantly lower than those in non-MCI group (IL-4: 0.875(0.143, 1.655)μg/L, 1.855(0.813, 2.723)μg/L; IFN-γ: 0.450(0.173, 1.163)μg/L, 1.160(0.483, 3.075)μg/L, all P<0.05). There was no significant difference in IFN- γ/IL-4, PSQI, HAMA 14 and HAMD 24 scores between MCI group and non-MCI group. IL-4 was positively correlated with the total score of MoCA( r=0.318, P<0.05), orientation( r=0.324, P<0.05)and delayed recall( r=0.368, P<0.01). The results of multivariate regression showed that IL-4 had significant effects on MCI in patients with chronic insomnia( B=2.161, OR=8.682, 95% CI=2.058~36.633, P=0.003). Conclusion:The cognitive function of chronic insomnia is closely related to serum IL-4 and IFN-γ, and serum IL-4 has a protective effect on cognition in chronic insomnia patients. Therefore, it can be speculated that cytokines may be an important pathophysiological link of cognitive change in chronic insomnia patients.

Objective:To investigate the clinical efficacy of neoadjuvant chemo-hormonal therapy(NCHT)followed by radical prostatectomy(RP) plus extended pelvic lymphadenectomy for very-high-risk locally advanced prostate cancer.Methods:The data of 327 cases of very-high-risk locally advanced prostate cancer treated in Renji Hospital Affiliated to Shanghai Jiaotong University School of Medicine, The Second Hospital of Tianjin Medical University, and The Third Affiliated Hospital of Sun Yat-sen University from December 2014 to July 2019 were retrospectively analyzed. Patients were divided into two groups according to treatment regimens: the RP group (direct RP + extended pelvic lymphadenectomy 4-6 weeks after the biopsy of prostate) and the NCHT group (4-6 cycles of NCHT prior to RP). There were 171 cases in RP group and 156 cases in NCHT group, respectively. In the RP group, the median age was 67 (ranging 44-83)years. The median PSA at diagnosis was 27.24 (ranging 4.55-207.00) ng/ml. Patients’numbers of clinical T 2, T 3a, T 3b, T 4 stage were 13, 85, 57, 16, respectively, and clinical N 1, N 0 stage were 33 and 138, respectively. Patients’numbers of ISUP grade groups of 1, 2, 3, 4, 5 were 5, 35, 41, 51, 39, respectively. In the NCHT group, The median age was 67 years, ranging 46-78 years. The median PSA at diagnosis was 72.09(ranging 4.08-722.95)ng/ml. Patients’ numbers of clinical T 2, T 3a, T 3b, T 4 stage were 11, 47, 58, 40, respectively, and clinical N 1, N 0stage were 76 and 80, respectively. Patients’numbers of ISUP grade groups of 1, 2, 3, 4, 5 were 1, 11, 33, 43, 68, respectively. At baseline, the NCHT group showed higher PSA, higher ISUP grade, and more advanced clinical stage at diagnosis( P<0.05). The PSA, pathological down-staging rate, and positive surgical margin rate as well as the biochemical recurrence free survival(bRFS)were compared between the two groups. Results:After radical prostatectomy, compared with the RP group, the NCHT group had a higher proportion of patients achieving PSA<0.2 ng/ml at 6-week postoperative follow-up ( P<0.001), a higher pathologic tumor stage down-staging rate ( P<0.001), a higher ISUP down-grading rate ( P<0.001), and a lower positive surgical margins rate ( P<0.001). In addition, 10.9% of the NCHT group achieved pT 0 or minimal residual disease in postoperative pathology exams. Eighty-three patients (48.5%) in the RP group and 125 patients (80.1%) in the NCHT group achieved undetectable PSA after surgery and entered further analysis for bRFS, which showed NCHT group had significantly longer bRFS (19.46 months vs. 6.35 months). NCHT significantly reduced the risk for biochemical recurrence in locally advanced prostate cancer patients( HR=0.278, 95% CI 0.198-0.390, P<0.001). Such a reduce in risk for biochemical recurrence was seen in all subgroups( P<0.001). Conclusions:NCHT might improve surgical outcomes as well as bRFS in very-high-risk locally advanced prostate cancer patients.

Objective To investigate the correlation between tumor-associated macrophages (TAMs) and the development of high risk human papilloma virus (hr-HPV)-related cervical cancer. Methods A total of 112 cases of cervical tissue were collected, including 16 normal cervical tissues, 55 cervical intraepithelial neoplasia (CIN) tissues and 41 squamous cervical cancer (SCC) tissues. The expression of CD163+ macrophages in the cervical tissues was detected by immunohistochemical method, and the results were analyzed in relation with the clinical data of the patients. Results Immunohistochemical analysis showed that the cell density of CD163+macrophages increased progressively with the increase in the tissue malignancy, in the order of normal cervical tissue, CIN I, CIN II-III, and SCC. Correlation analysis revealed a positive relationship between CD163 + macrophage density and tissue malignancy (P=0.000). The density of CD163 + macrophages was significantly upregulated in HR-HPV-positive SCC tissue (P<0.05). CD163+ macrophages were positively correlated with cervical lymph node metastasis (P=0.005) and FIGO stage (P=0.004) of SCC. Conclusion The expression of CD163+macrophages is positively correlated with malignant transformation of cervical tissues, and hr-HPV infection is significantly correlated with CD163 expression level in the macrophages. CD163+ macrophages can be used as predictors of the occurrence and progression of cervical cancer caused by hr-HPV infection.

Objective To evaluate the relative safety of different ventilation methods regarding mortality and rates of complication,on neonatal respiratory distress syndrome (NRDS).Methods Network Meta-analysis was used to collect data on randomized controlled trials of pulmonary ventilation strategies in preterm infants with a mean gestational age of less than 32 weeks.Diagnostic criteria on NRDS were published in the PubMed,Cochrane,Web of Science,EBSCO,and Springer Link databases from January 1986 to June 2018.Revman 5.3 software was used to evaluate the quality of studies,based on the Cochrane quality assessment tool.Data were analyzed by Bayesian and frequency methods,using both Win BUGS 1.4.3 and STATA 13.0 software.Safety of different ventilation strategies for NRDS mortality and complications would include intraventficular hemorrhage (IVH),patent ductus arteriosus (PDA) and retinopathy of prematurity (ROP) and were evaluated.Counted data was displayed by OR and 95％CI.Results A total of 31 RCTs were included in this paper,including 5 827 preterm infants and 11 ventilation strategies.There were no statistically significant differences appearing in 11 ventilation strategies on mortality,PDA or ROE IVH results were reported in 28 studies.Compared with nasal intermittent positive pressure ventilation (NIPPV),both high-frequency oscillation ventilation (HFOV) (OR=3.33,95％ CI:1.08-16.67,P＜0.05) and synchronized intermittent mechanical ventilation (SIMV) (OR=8.22,95％CI:1.25-29.44,P＜0.05) schemes seemed to have increased the risk of IVH in preterm infants with NRDS.NIPPV appeared the optimal ventilation strategy in the rankings of cumulative probability.Results on clustering showed that NIPPV was probably the best ventilation strategy for children with NRDS after considering the orders of IVH,PDA and ROP on mortality,respectively.However,HFOV,IMV,and SIMV did not seem to be the ideal ventilated strategies.Conclusions Most of the clinical decision makers might prefer using NIPPV in the treatment of children with NRDS through mechanical ventilation systems to reduce both the incidence and death caused by IVH,PDA and ROP.It was not recommended to use HFOV,SIMV and IMV in treating NRDS with gestational less than 32 weeks.We suggested that larger numbers of multi-center RCTs ba carried out to make the above conclusions more convincing.

Objective To investigate the correlation between tumor-associated macrophages (TAMs) and the development of high risk human papilloma virus (hr-HPV)-related cervical cancer. Methods A total of 112 cases of cervical tissue were collected, including 16 normal cervical tissues, 55 cervical intraepithelial neoplasia (CIN) tissues and 41 squamous cervical cancer (SCC) tissues. The expression of CD163+ macrophages in the cervical tissues was detected by immunohistochemical method, and the results were analyzed in relation with the clinical data of the patients. Results Immunohistochemical analysis showed that the cell density of CD163+macrophages increased progressively with the increase in the tissue malignancy, in the order of normal cervical tissue, CIN I, CIN II-III, and SCC. Correlation analysis revealed a positive relationship between CD163 + macrophage density and tissue malignancy (P=0.000). The density of CD163 + macrophages was significantly upregulated in HR-HPV-positive SCC tissue (P<0.05). CD163+ macrophages were positively correlated with cervical lymph node metastasis (P=0.005) and FIGO stage (P=0.004) of SCC. Conclusion The expression of CD163+macrophages is positively correlated with malignant transformation of cervical tissues, and hr-HPV infection is significantly correlated with CD163 expression level in the macrophages. CD163+ macrophages can be used as predictors of the occurrence and progression of cervical cancer caused by hr-HPV infection.