Objective To investigate the influence of the high-flux hemodialysis (HFD) on inflammatory factors [C-reactive protein (CRP),interleukin-6 (IL-6),tumor necrosis factor-α (TNF-α)] and serum brain natriuretic (BNP) in patients before and after hemodialysis.Methods Fifty patients with MHD were enrolled in our hospital in 2013 and divided randomly into HD group (n =25),and HFD group (n =25).Serum CRP,IL-6,TNF-α,and BNP were measured in patients before and after the first hemodialysis session and after the treatment for 6 months later.Results Serum CRP,IL-6,and TNF-α in HD group after the first dialysis session and after the treatment for 6 months were statistically insignificant (P ＞0.05).In HD group,serum BNP was decreased after the first hemodialysis session and the treatment for 6 months later,the decreases was statistically significant (P ＜0.01 or P ＜0.05).In HFD group,serum CRP,IL-6,and TNF-α were decreased after the first treatment,the decreases were statistically insignificant (P ＞ 0.05).However,after the 6 mouths,the decreases were statistically significant (P ＜ 0.05).Serum BNP was decreased after the first hemodialysis session and the treatment for 6 months later,the decrease was statistically significant(P ＜ 0.01).Between two groups,the different time-point and the interaction of two groups and the different time-point,the discrepancy above them were statistically significant (P ＜0.01).Conclusions The high-flux hemodialysis can eliminate more inflammatory factors such as CRP,IL-6,TNF-α,and serum BNP in patients with uremia.Serum BNP Helps to adjust the dry weight in time,relieve cardiac load and reduce the incidence of heart failure.High-flux hemodialysis is one of the ways of treatment that reduce the morbidity of the heart failure.It can improve prognosis of the patients with MHD.

Objective To explore the clinical efficacy and safety of mifepristone combined with misoprostol in the treatment of missed abortion.Methods According to the digital table,184 patients with missed abortion were randomly divided into study group and control group,92 cases in each group.The control group was given oral diethyl-stilbestrol underwent curettage,while the study group was given mifepristone combined with misoprostol therapy under-went curettage.The clinical therapeutic effect and adverse reaction were compared between the two groups.Results The effective rate of the study group after treatment was 95.65%,which was significantly higher than 86.96% of the control group,the difference was statistically significant(χ2 =4.38,P <0.05).Compared with the control group,in the study group,the operative time[(7.22 ±1.11)minute]observed significant relationship,intraoperative hemorrhage volume[(35.68 ±5.62)milliliter]decreased significantly,pregnancy natural rate of discharge(81.52%)was obvi-ously improved,artificial abortion syndrome incidence rate(3.26%)and again the rate of curettage(5.43%)was obviously decreased,and the differences were statistically significant(t =16.10,12.52,χ2 =7.84,8.87,15.93,all P <0.05).The clinical adverse reaction rate in the study group was 16.30%,which was significantly lower than 29.35% in the control group,the difference was statistically significant(χ2 =4.44,P <0.05).Conclusion Mife-pristone combined with misoprostol plays a complementary role,can improve the treatment of missed abortion and has good operation condition.It also can reduce the adverse reactions,complications and pain of patients.It is worth of clinical promotion.

Based on the references of relevant statistical data, the implementation of the overall situation of the basic medical insurance system in Henan were compared. The sample cities and counties of basic medical insurance data were analyzed through the field survey. Using empirical analysis methods to discuss the necessity and feasibility of integration between new rural cooperative medical system and urban residents medical insurance, put forward the integration path and propose related suggestions.

Objective To explore the effects of murinecytomegalovirus(MCMV)infected mouse embryonic fibroblasts(MEF)on the proliferation and activation of regulatory T cell in vitro. Methods A co-culture system of T cell and MCMV infected MEF(T-MEF MCMV)was established.The viral load of supernatant was determined by plaque assay.The proliferation of T cells was observed with cell counting.The proportion of CD4+ CD25+ cells was measured by flow cytometry.The levels of Foxp3 protein were measured by Western blot.Reverse transcription polymerase chain reaction(RT-PCR)assay was utilized tO determine whether MCMV infection of MEF influenced the level of transforming growth factor(TGF)-β mRNA.The level of TGF-β protein in supernatant was measured by double-antibody sandwich enzyme linked immunosorbent assay(ELISA).The difference between T-MEF MCMV group and control group was assessed by one-way ANOVA.Results When T cells were co-cultured with MCMV infected MEF for 1 day and 3 days,the viral load in supernatant decreased.But when co-culture lasted for 6 days,the antiviral effect obviously diminished,as the viral load[(5.58±0.67)×105 PFU/mL]of the experimental group showed no statistic difference with MEF MCMV control group[(6.05±0.34)×105PFU/mL].When co-cultured with MCMV infected MEF for 3 days,T cell increased from pre-culture level of[(2.02±0.05)×106/mL]to(2.25± 0.13)×106/mL(P＜0.05).But when co-culture lasted for 6 days,the number of T eelI returned to (2.08±0.14)×106/mL,which had no statistic difference with that of co-culture for 3 days system. Both the expressions of Foxp3 protein and the proportion of CD4+ CD25+ Foxp3+ Treg cells in T-MEF MCMV group were up-regulated as the infection time extended,which were two times and three times of the control group level,respectively.The mRNA level(A value)of TGF-β in MCMV infected MEF increased from baseline of 1.09±0.13 to 3.15±0.54 on day 3 after infection.The expression of TGF-β in supernatant 3 days after infection was(3.85±0.32) μg/L,which was significantly higher than that before infection[(1.74±0.14)μg/L,P＜0.05].Conclusions Activated T cells have antiviral effect.However,the function of T cells is rapidly inhibited after activation,which may be due to the expression of Foxp3 mRNA induced by MCMV infected MEF and increased CD4+ CD25+ Treg proportion of the co-cultured T cells.TGF-β level is significantly increased after CMV infection,which may be an important mechanism of Treg proliferation.MCMV may manipulate Treg to evade specific immune elimination and,as a result,to cause CMV replication.

Objective:To observe the influence of the “Warming yang and Nourishing qi” acupuncture treatment in the serum interleukin-12 (IL-12) and interleukin-18 (IL-18) expression levels,in the rats with experimental autoimmune myasthenis gravis (EAMG) and to analyze the mechanism of acupuncture treatment to myasthenis gravis (MG).Methods: AchRα1 129-145 peptide fragments were used to immune the female Lewis rats to establish EAMG rat models.30 successful EAMG model rats were randomly selected and divided into acupuncture treatment group ,drug control group and model control group ( n=10 );the unmodelled 10 rats purchased at the same period were used as blank control group.The rats in acupuncture treatment group were treated by “Warming yang and Nourishing qi” acupuncture treatment,30 min/time,once a day,7 d for a period,1 d rest between two periods,two periods of continuous treatment.The rats in drug control group were treated by the pyridostigmine lavaged treatment , 18.5 mg/( kg · d ) , continuous treatment of 15 d.The rats in the other two groups had no any special treatment.After treatment,the blood samples from the vein of tail root of the rats in 4 groups were collected ,and the serum expression levels of IL-12 and IL-18 were detected with ELISA method.Results:After treatment,compared with model control group ,acupuncture treatment and drug control group the serum IL-12 and IL-18 expression level of the EAMG rats were significantly decreased ( P<0.05 or P<0.01 ).Compared between acupuncture treatment group and drug control group showed that ,the serum IL-12 and IL-18 expression level of the EAMG rats were no significant difference ( P>0.05 ).Conclusion:“Warming yang and Nourishing qi” acupuncture treatment can reduce the serum IL-12 and IL-18 expression levels of EAMG rats to treat MG ,and the effect of treatment was similar to pyridostigmine.

Objective To investigate the influencing factors of the results of liver function tests.Methods The clinical data of 357 cases of outpatient examination for liver function were retrospectively analyzed.According to the results of the examination,they were divided into two groups,178 cases in the control group (normal liver func-tion);179 cases in observation group (abnormal liver function).The influencing factors of liver function test results were analyzed.Results In the observation group,the gender,marital status,age,culture degree,BMI,sleep time com-pared with the control group,the differences in marital status were statistically significant(all P <0.05),the liver function abnormal rate of male was 58.66%,which was higher than 41.34% of female(χ2 =14.139,P <0.05).The detection rate of abnormal liver function of the people with age over 40 years old was 55.87%,which was higher than 44.13% of the people with age under 40 years old (χ2 =8.495,P <0.05).The detection rate of abnormal liver func-tion of the crowd in high school level and above was 45.25%,which was lower than 54.75% of the crowd under the high school(χ2 =5.685,P <0.05).The detection rate of abnormal liver function of the crowd with BMI≥24kg/m2 was 62.57%,which was higher than 37.43% of the crowd with BMI <24kg/m2 (χ2 =5.731,P <0.05).The detec-tion rate of abnormal liver function of the crowd with sleep time≥8h was 53.07%,which was lower than 46.93% of the crowd with sleep time <8h (χ2 =18.011,P <0.05).There was no significant difference in the detection rate of abnormal liver function of marriage between the two group(P >0.05).Conclusion The gender,age,culture degree, BMI,sleep time have impact on the liver function test,and the bad eating and living habits should be reasonable corrected,the physical exercise should be strengthened,and the quality of life should be constantly improved.

AIM: To investigate whether efficient production of neuron-like cells from embryonic stem cells(ESCs) can be indued by astrocyte-conditioned medium(ACM) in vitro.METHODS: Based on the 4-/4+ protocol established by Bain,two groups were studied: ATRA group,ATRA with ACM group.ESCs were induced into neuron-like cells by means of three-step differentiation in vitro.The totipotency of ESCs was identified by the observation of cells' morphology and the formations of teratoma in immunocomprised mice.The cell differentiation was evaluated continuously by detection of the cellular specific markers of neural stem cell, neurons and astrocytes such as nestin,NF-200,NSE and GFAP using immuno-histochemistry assay.RESULTS:(1) The ESC-D3 cells kept the ability of differentiation into cellular derivations of all three primary germ layers after continuous passage culture.(2) The ratio of NF-200 and NSE positive cells in the cells induced by ATRA with ACM was higher than that in the cells induced by ATRA only.(3) Finally,the positive rate of the neuron-like cells was up to 73.5% in the group induced by ATRA with ACM.CONCLUSION: The ESCs are induced into neuron-like cells with high purity and efficiency by ATRA with ACM.

Aim To establish a highly purified,active and prac-tical extract and primitive culture method for rat brain microvas-cular endothelial cells ( BMECs) for providing materials for con-struction in vitro blood-brain barrier ( BBB) model. Methods Cerebral cortex of 1-2 week SD rats was collected,and successive digestion with typeⅡcollagenase and collagenase/dispase, sieve filtration and then twice gradient centrifugation in 20% BSA and 44% Percoll condition were used to obtain brain microvascular section. After that brain microvascular section was seeded in cul-ture bottle and then primarily cultured. Inverted microscope and factor-VIII relative antigen immunostaining methods were used for cellular morphological observation and identification. Results BMECs climbed out the vessel segment and proliferated with ad-herence after 2 h in vitro culture,and they became typical peb-bles structure after further 3-4 d culture. Morever, factor-VIII relative antigen immunostaining identified that expression for the endothelial cells was positive,cytoplasm was brown and positive cells account for more than 99%. Conclusion Rat BMECs with high purity could be extracted and cultured by using the above methods,and it has great potential for construction in vitro BBB model and in depth studies on biological characteristics and func-tions of BMECs.

To establish a method of directional differentiation and efficient production of neurons from embryonic stem cells (ES cells) in vitro, based on the 4-/4+ protocol described by Bain, a new method was established to induce ES cells differentiating into neurons by means of three-step differentiation using all-trans retinoic acid (ATRA) combined with astrocyte-conditioned medium (ACM) in Vitro. The totipotency of ES cells was identified by observation of cells' morphology and formations of teratoma in immunocompromised mice. The cells' differentiation was evaluated continuously by the detection of the specific cellular markers of neural stem cells, neurons and astrocytes, including nestin, NSE and GFAP using immunohistochemistry assay. The NSE positive cells' ratio of the differentiated cells was determined by flow cytometry. It was found that the transparent circular clusters surrounding embryoid bodies induced with combining induction protocol formed just after 24 h and gradually enlarged later. This phenomenon could not be observed in EBs induced only by ATRA. The NSE positive cells' ratio in the cells induced with ATRA and ACM was higher than that of the cells induced by ATRA at different time points of differentiation, and finally reached up to 73.5% among the total differentiated population. It was concluded that ES cells could be induced into neurons with high purity and yield by means of inducing method combining with ATRA and ACM.
Astrocytes/*cytology ; Cell Differentiation ; Cells, Cultured ; Embryo, Mammalian ; Neurons/*cytology ; Stem Cells/*cytology ; Tretinoin/pharmacology

Objective To validate a new standardized training program for urological surgeons to improve their laparoscopic surgical skills. Methods The laparoscopic surgical training program was carried out by using the traditional mechanical simulators and animal models. Thirty-three trainees participated the urological laparoscopic surgical training program. Initially, the novices were assigned to practise basic laparoscopic skills step by step on the simulator with fixed trocar positions. After a period of basic training, they were allowed to practise on animal models for some particular proce-dures. Results All trainees (33/33, 100.0%) participated were able to perform all basic techniques skillfully and completed laparoscopic anastomosis accurately after the training. The time required for performing the partial nephrectomy, dismembered pyeloplasty and ureteral reimplantation on animal models declined from 64.0±18.4, 127.54±17.5 and 75.84±11.6 min at the beginning to 30.94±3.8, 65.2±7. 5 and 37.7±7.2 min after practicing these procedures 8 times (P<0.01). They could un-derstand the crucial procedures of the laparoseopic surgeries after 6 to 8 special trainings on animal models. Fifteen trainees (15/33, 45.5%) had started to carry out laparoscopic surgeries after-finish- ing the training program. Conclusions Our program enables the participants to improve their techniques in complicated laparoscopic surgeries. The challenging parts of reconstructive laparoscopic surgeries such as laparoscopic dismembered pyeloplasty can be taught by using animal models. This program could be incorpo-rated easily by all urological departments developing a laparoscopie surgical training program.