Objective To observe the effects of low intensity electromagnetic fields (LIEMFs) in promoting the reconstruction of full skin loss wounds grafted with human epidermal stem cells (ESCs). Methods Fifty nude mice aged 7 to 8 weeks with full skin loss wounds were equally divided into 3 experimental subgroups ( 1 Hz, 10Hz and 50Hz) and two control groups (a cell suspension control group and a blank control group) , with ten mice each. In the 3 experimental subgroups and the cell suspension control group, ESCs separated from human foreskin and cultured in vitro were grafted to the wounds using collagen sponge scaffolds. The experimental subgroups were then stimulated with an LIEMF (magnetic field intensity 5mT) at the appropriate frequency for 30min/day for 15 days. The blank control group was put under the same conditions without the cell suspension and LIEMF. The healing rates of the wounds were observed, and tissue slices were stained and observed under a light microscope. The inner structure of the regenerating skin was observed using transmission electron microscopy. Results The ESCs were successfully grafted. A few human integrin β1 positive stained cells appeared in the regenerating skin. The average healing rates in the experimental subgroups were significantly superior to those of the control groups. Well differentiated epidermis and dermis could be seen in the regenerating skin in all of the experimental groups. The epidermis had more cell layers and was thicker than in the control groups. More desmosome, hemidesmosome and keratin filaments were seen among the epidemic cells of the experimental groups. Conclusions LIEMF promotes the healing of full skin loss wounds grafted with ESCs in nude mice, and can promote complete repair of skin defects and the regeneration of skin function.

BACKGROUND:Previous studies have shown that a certain dose of acidic fibroblast growth factor can promote skeletal muscle satelite cel proliferationin vitro. OBJECTIVE:To investigate the effects of transfection with acidic fibroblast growth factor by electroporation on growth, proliferation and differentiation of skeletal muscle satelite cels. METHODS: Skeletal muscle satelite cels were cultured and purified, and then transfected with plasmid pSectag-GFP-aFGF by electroporation. The expression of green fluorescent protein was observed under fluorescence microscope, and the transfection efficiency was calculated. After transfection, cel cycle was analyzed by flow cytometry to draw the growth curve of skeletal muscle satelite cels. Western blot assay was employed to measure protein level of acidic fibroblast growth factor. RESULTS AND CONCLUSION: (1) Immunocytochemistry detection: The skeletal muscle satelite cels were positive for a-sarcomeric actin. (2) Transfection efficiency: At 12 hours after transfection with pSectag-aFGF, several cels showed green fluorescence, and the green fluorescent expression reached the peak at 72-96 hours after transfection with a positive rate of about 90%. (3) Cel cycle: After electrotransfection, the proportion of cels at S phase in the electroporation group was higher than that in the control group (P < 0.05). (4) Cel growth curve: At 3 days after electrotransfection, the cels entered logarithmic growth phase but the proliferation slowed down at 5 days. (5) Differentiation capacity: There were fewer myotubes and aging cels in the electroporation group than the control group. (6) Western blot assay: Acidic fibroblast growth factor protein was highly expressed in the cels transfected with target gene detected by western blot assay. These findings indicate that by using electroporation method, acidic fibroblast growth factor can be transferred into skeletal muscle satelite cels and have a high-efficiency and long-term expression, which can promote the proliferation of skeletal muscle satelite cels and inhibit formation of myotubes.

Objective To dynamically monitor the changes of peripheral blood lymphocyte subsets of renal transplant recipients and investigate the relationship between lymphocyte subsets with infection and rejection.Methods The clinical data of allogenic kidney transplantation recipients and living relative donors in the Department of Kidney Transplantation of the First Affiliated Hospital of Zhengzhou University were prospectively collected from June 2015 to December 2016.The data of lymphocyte subsets and other related indexes were obtained from renal transplant recipients and relatives of the same period.Results Sixty-four cases of living-relative donors and 351 cases of renal transplant recipients were enrolled in this study,and the recipients were divided into 3 groups:infection group (67 cases),acute rejection group (46 cases),and stable group (238 cases),according to the diagnostic criteria.There was significant difference in the concentration distribution of lymphocytes between the stable group and the control group (P<0.05).The stable frequency distribution range of the stable group was as follows (cells/μL):Lym (1 000-1 500),T (<1 500),CD4+ (<1 000),CD8+ (<1 000),B (<300),NK (100-300),CD4+/CD8+ (0.5-1.0).The number of Lym,T,CD4+,CD8+,NK and B cells in the preoperative patients was less than that in the healthy population (P<0.05);The number of Lym,T,CD4+, CD8+,B and NK cells was gradually decreased in the postoperative infection group,which was less than that in the stable group (P<0.05).After treatment the indicators gradually restored to the level in the stable group level;the number of T,CD4+,CD8+,B cells was highly correlated with infection.The number of T and CD4+ cells,and CD4+/CD8+ ratio were significantly increased in acute rejection group as compared with the stable group,and gradually decreased after the rejection was reversed.The number of T,CD4+,CD8+ cells was highly correlated with rejection.Lymphocyte subsets had a predictive effect on infection and rejection of recipients,and CD4+ cell count and CD4+/CD8+ ratio were independent risk factors.Conclusion The monitoring of lymphocyte subsets has an important clinical value in the evaluation of immune status and individual treatment of renal recipients.

Objective To investigate the role of BMSCg on enhancing the implant survival and bacheal epithelium regeneration. Methods After transplanted with cryopreserved 2 weeks and 6 weeks allocraft, PHK-26 labeled 3-5 passage BMSCs were injected into the recipient rats via tail vein. Rats in the control groups were injected with the same amount of PBS.We observed the histology of the transplanted trachea including epithelium growth and regeneration, and the PKH-26 fluorescence levels at the para-anastomotic trachea to evaluate the role of BMSC transplantation on the epithelium regeneration. Results Rats from BMSCs injection group survived a long period. Histological observation showed that the tracheal lumen was covered by psudo-striated ciliated columnar epithelium. The cartilage structure was intact. There are no signs of denaturation and necrosis. In the PBS injection group, epithelium regeneration is better in PBS-6-week group than PBS-2-week group. The longest survival time in PBS-6-week group was 32 days, whereas it was 10 days in PBS-2-week group. In BMSCs injection group, rats in BMSC-6-week groups survived longer than 8-week group(12 rats were terminated at 1 week, 4 weeks and 8weeks as planned). There was one rat who survived and were terminated at the designated 8 weeks time point (there were 8regenerated epithelium was similar in the two BMSC transplanted groups. PKH-26 labeled BMSCs migrated to the implant site and showed red fluorescence, with most red fluorescence shown at the anastomotic part. Conclusion BMSCs can migrate to the impaired tissue to repair it. BMSCs may exert their reparation function via enhancing epithelium regeneration.

Objective To investigate the status of living relative kidney transplantation, and the clinical effects and social significances of kidney transplantation between spouses. Methods We retrospectively collected the clinical data of the department of kidney transplant of the First Affiliated Hospital of Zhengzhou University, from January 2011 to December 2013. The spouse group as group 1, the age and sex of doners were taken into account,the siblings with the similar age of the same period were enrolled in group 2. Then the postoperative recoveries of the two groups were compared. Considering the current social status, particularly the shortage of donor kidneys, the clinical, social and family significances of kidney transplantation between spouses were analyzed. Results Twelve cases of spouses in group 1, 8 cases of siblings in group 2 , the differences of donor and recipient age of the two groups were 0.33 ± 0.98 years and 2.29 ± 7.23 years, respectively. The human major histocompatibility complex antigens (HLA) was less than three in group 1, and was greater than or equal to three in group 2. The changes of serum creatinine and urea nitrogen were analyzed. No significant differences of serum creatinine and downward trend of blood urea nitrogen were observed between two groups (P = 0.84, P = 0.79). Conclusion The kidney transplantation between spouses has good clinical efficacy and great social significance, improving the status of the shortage of donor kidney and contributing to family harmony. The renal transplantation between spouses has obvious advantages and need further promotion.

Objective To analyze the clinical and laboratory characteristics of the ischemic stroke in patients with non-valvular atrial fibrillation (AF), and to provide evidence for the prevention of ischemic stroke. Methods A total of 198 patients with ischemic stroke were chosen and divided into two groups:with AF (71 patients)/and without AF (127 patients) groups. Clinical data and biochemical markers were collected and compared in two groups. CHADS2 and CHA2DS2-VASc score systems were used to determine the risk levels in patients with AF. Finally, related risk factors of ischemic stroke with AF were determined and analyzed. Results The values of age, length of hospital stay, the hypertention history, heart rate and plasma homocysteine (Hcy) were significantly higher in the with-AF group than those in the without-AF group ( P <0.05). The levels of total cholesterol (TC), triglyceride (TG) and very low density lipoprotein cholesterol (VLDL-C) were sig?nificantly lower in the with-AF group than those of the without-AF group (P<0.05). CHA2DS2-VASc scores reached to the moderate-to-high risk level in the with-AF group. Multiple-factor logistic regression analysis showed that age and heart rate were the independent risk factors of the ischemic stroke in patients with non-valvular AF. ROC analysis indicated that age (AUC=0.761, cut-off point=72.50 years old) and heart rate (AUC=0.612, cut-off point=76.50 bit/min) had predictive and di?agnostic value for the ischemic stroke in patients with non-valvular AF. The age of these patients had the best sensitivity (70.4%) and specifity (71.1%), and the cut-off point of which was 72.50 years old. Conclusion The characteristics of isch?emic stroke in patients with non-valvular AF includes older age, faster heart rate, higher CHA2DS2-VASc scores and higher Hcy level.

Objective To investigate the effect of icariin on renal ischemia-reperfusion injury (IRI) and the action mechanism.Methods SD rats were divided into 3 groups.The model of unilateral renal IRI was established in SD rats,and Icariin (100 mg/kg) was orally administrated by gavage daily from 2 days before operation to 12 days after operation (icariin group).The vehicle of icariin was administrated to IRI model rats as control group,while in the sham-operation group the renal pedicel was only dissociated without treatment.Body weight and kidney function were monitored within 14 days after reperfusion.The kidney was harvested at 24 h after reperfusion,and then malonaldehyde (MDA) and activity of reactive oxygen species (ROS) scavenger enzymes were examined.Histopathological changes were observed at postoperative day (POD) 3 and 14.Results At POD 3,7,11 and 14,the creatinine clearance rate was significantly higher in icariin group than in control group (P＜0.01). Icariin group had significantly lower Paller scores which indicated tubules injury than in control group at day 3 after reperfusion (P＜0.01 ).In icariin group,MDA level was obviously decreased at 24 h after operation.Compared to control group,icariin group had statistically higher activity of glutathion reductase (GR),catalase and superoxide dismutase (SOD),as well as higher level of reduced glutathione (GSH) (P＜0.05).Conclusion Icariin has protective effects on renal IRI,and can promote recovery of kidney function. Icariin can reduce oxidative stress through increasing activity of ROS scavengers.

Objective To analyze the clinical effectiveness of using marginal donor kidney in living kidney transplant. Methods From November 2005 to June 2011,274 cases of living kidney transplant were performed in the First Affiliated Hospital of Zhengzhou University.The cases were divided into the marginal donors group ( Donor ages over 60 years old,suboptimal renal anatomy or physiology) of 66 cases and standard donors group of 208 cases.The clinical data were retrospectively analyzed.The criteria of marginal donors were as follows:36 cases of donors with age over 60 yrs (6 cases with renal cysts and 1 case with renal calculus),22 cases of renal cysts ( with diameter range from 4 mm to 40 mm ),4 cases of renal calculus (with diameter range from 3 mm to 6 mm),4 cases of low GFR (under 35 ml/min.The mean recipients' serum creatinine before surgery and after surgery on day 3,day 7,month 1,month 3,month 6,month 12,related complications,the rate of acute rejection and delayed graft function,1 year,3 year recipient/kidney survival were compared between the 2 groups,respectively. Results The serum creatinine levels in the marginal donor group and standard donor group were (242.7 ± 132.2 vs 185.6 ± 148.4) and ( 156.7 ±86.8 vs 122.2 ± 136.8 ) on day 3,day 7 respectively ( P ＜ 0.05 ).Nevertheless,there were no significant differences between the 2 groups in recipients' serum creatinines before surgery and after surgery on month 1,month 3,month 6,month 12,peri-operative complications,the rate of acute rejection and delayed graft function,1 year,3 year recipient/kidney survival (P ＞ 0.05). Conclusions Healthy old donors and donors with renal cyst (the diameter of renal cysts under 40mm) should not be the barriers to organ donation.To those living donors with low GFR,we should consider of donor age,donor/recipient body weight,donor/recipient body surface area and whether we could deal with the problem by surgical operation.Donor with renal calculus should be carefully evaluated.

Objective To analyze clinical outcome of kidney transplantation from elderly livingrelated donors. Methods The clinical data of living-related kidney transplantation between November 2005 to June 2011 in the First Affiliated Hospital of Zhengzhou University were retrospectively analysed.The patients were divided into elderly donor kidney transplantation group (elderly group,36 cases) and non-elderly living related donor kidney transplantation group (nonelderly group,208 cases).The mean recipients' serum creatinines before and 3 days,7 days,1 month,3 month,6 month,12 month,24 month,36 month after surgery,peri-operation complications,the incidence rate of acute rejection,calcineurin inhibitor (CNI)-induced renal toxicity,delayed graft function(DGF),1 year and 3 year recipient/kidney survival were compared respectively between the two groups. Results The serum creatinine levels were higher in elderly group than in non-elderly group at 3 days,7 days,1 month after surgery [(245.2±135.2)μmol/L vs.(185.6±148.4)μmol/L,(150.5±86.8)μmol/L vs.(122.2± 136.8)μmol/L,(140.6±42.5)μmol/L vs.(117.8±33.2)μmol/L,t =84.07,31.90,21.54; all P =0.000].In addition,the incidence rate of CNI induced renal toxicity was higher in the elderly group than in non-elderly group [22.2％(8/36)vs.1.9％(4/208),x2=27.04,P=0.000].Nevertheless,there were no significant differences between the two groups in recipients' serum creatinines before and 3 month,6 month,12 month,24 month,36 month after surgery,in peri-operation complications,the incidence rate of acute rejection,delayed graft function,and in 1 year and 3 year recipient/kidney survival (all P ＞ 0.05). Conclusions With stringent screening and overall assessment of elderly donors,healthy elderly donors should not be barriers to organ donation.Renal toxicity of CNI agents should be carefully monitored in recipients of elderly donor.

BACKGROUND: Donation after cardiac death (DCD) is becoming the main source of organs for transplantation. Donor and recipient gender may play an important role in preoperative evaluation and recipient selection of transplantation. OBJECTIVE: To analyze the effect of donor and recipient gender on the recovery of renal function after DCD renal transplantation, and to further guide the selection of recipients. METHODS: The clinical data of recipients and donors of DCD kidney transplantation performed at the First Affiliated Hospital of Zhengzhou University from July 2012 to March 2015 were analyzed retrospectively. According to donor and recipient gender, the donors and recipients were divided into four groups: group A (male donor, male recipient), group B (male donor, female recipient), group C (female donor, male recipient), group D (female donor, female recipient). The renal function of the recipients was recorded at 1 and 2 weeks, 1, 3, 6, and 12 months postoperatively to compare the effect of donor and recipient gender on the recovery of renal function. RESULTS AND CONCLUSION: The serum creatinine level in the groups A and B was lower than that in the groups C and D at postoperative different time points, and there was a significant difference at 3, 6 and 12 months postoperatively (P < 0.05). The estimated glomerular filtration rate in the groups A and B was significantly higher than that in the groups C and D at postoperative different time points (P < 0.05). The serum creatinine level in the group C was significantly higher than that in the other three groups at postoperative different time points (P < 0.05). The serum creatinine level in the group A was significantly higher than that in the group B at 3, 6 and 12 months postoperatively (P < 0.05). The glomerular filtration rate in the group A was significantly higher than that in the group C at postoperative different time points (P < 0.05). The glomerular filtration rate in the group B was significantly higher than that in the group D at 12 months postoperatively (P < 0.05). These results suggest that the male donors show superior renal function to the female donors. The postoperative recovery of renal function is relatively slow when the male recipients receive a kidney of female donors. The female recipients with lower body surface area who receive the kidney of elderly male donors with poor renal function can achieve favorable clinical effects. Therefore, the gender should be used as a reference index for selecting potential recipients.