Objective To evaluate the feasibility of the diagnosis of the early immunologic rejection after xeno-islet transplantation by MR imaging enhanced with superparamagnetic iron oxide(SPIO)marking CD4+T cell antibody.Methods Two thousand neonatal porcine islets(NPI)were transplanted under the left renal capsule of BALB/C nude mice.When the grafts could be observed bv MRI.107 human PBMC was intraperitoneal injected to nude mouse models to reconstitute the human immunologic system,20 mice were reconstituted.Before and 3,7,14 days after reconstitution of human immunologic system on BALB/C nude mice,MRI imaging Was performed half an hour after intravenous injection of nano-immunomagnetic beads via vena caudalis to observe the grafts'MRI signal.BALB/C nude mice were sacrificed after MRI scanning immediately,the histopathologic examination was assessed on grafts,the results were compared with MRI results.And calculate the sensitivity,specificity,Youden index number and coincidence of the MRI for immunologic rejection.Results Grafts can be observed bv MRI 3 weeks after islet cell transplantation (before immunologic rejection modeling),there is no abnormal MRI signal detected in nude mice'graft region after mierobeads injected.Seven days after building of immunologic rejection model,MRI hypo-signal in graft site is shown in the T2 WI sequence after nano-bioprober injected.Histopathologic assessments were employed on grafts in nude mice immediately(HE and immunohistochemistry staining),the results shown that there are a lot of T lymphocyts infiltrated in graft region.implying the occurrence of immunologic rejection.And the sensitivity,specificity,Youden index number and coincidence is:(72.96±0.24)%,100%,0.73±0.24,(88.46±0.13)%respectively.The correct Kappa between the MRI and the imunohistochemistry staining was 0.76.Conclusion The cellular immunological rejection to xeno-islet grarts can be assessed with nano-bioprobe with anti-CD4+ antibody MR imaging,real time,and noninvasively.

Objective To evaluate the effectiveness and safety of mouse nerve growth factor in treating dysphagia in patients with nasopharyngeal carcinoma after radiotherapy.Methods Fifty-eight post-radiotherapy nasopharyngeal carcinoma patients with dysphagia were randomly divided into an observation group and a control group.Both groups received routine treatment,but the observation group was additionally injected with mouse nerve growth factor intramuscularly every day for four weeks.Before and after the 4 weeks of treatment,both groups were evaluated using Kubota's water drinking test,videofluoroscopy and the brief version of the WHO's Quality of Life scale.Results After 4 weeks,the patients in the observation group displayed significantly greater improvement in swallowing compared with the control group.There was a significant difference in the groups' average scores on the drinking water test and in the videofluoroscopy results.Moreover,the patients in the observation group had significantly higher quality of life scores than those in the control group,on average.Conclusions Mouse nerve growth factor may have a rapid and safe therapeutic effect on dysphagia induced by radiation.No obvious adverse reactions were observed.

Objective: To observe the therapeutic efficacy of sinew-bone balancing manipulation plus exercise therapy in treating postures of primary school students with upper crossed syndrome (UCS).Methods: Sixty pupils with UCS were divided into an exercise group and a combination group using the random number table method, with 30 cases in each group. The combination group received treatments of sinew-bone balancing manipulation plus exercise therapy, while the exercise group received exercise therapy alone. The two groups received interventions once every other day, for a total of 1 month. The sagittal static posture assessment total score, forward head angle (FHA) and forward shoulder angle (FSA) were compared before and after treatment; the sagittal static assessment total score, FHA and FSA were compared between the exercise group and the combination group. Results: Before treatment, there were no significant differences comparing the sagittal static posture assessment total score, FHA and FSA between the two groups (all P>0.05); after treatment, the sagittal static posture assessment total score, FHA and FSA decreased in the two groups, with intra-group statistical significance (all P<0.01), and were lower in the combination group than in the exercise group, with inter-group statistical significance (all P<0.01). Conclusion: Sinew-bone balancing manipulation plus exercise therapy can notably improve the FHA and FSA and reduce the sagittal static posture total score in pupils with UCS, so as to correct the bad postures and adjust UCS physique. It can produce more significant efficacy compared with exercise therapy alone.

Objective To study the iodine nutritional status of population living in Tibetan pastoral areas,in order to provide a scientific basis for prevention and control of iodine deficiency disorders.Methods Drinking water samples were collected to test iodine content in agricultural town(Kajiaman) and pastoral area(Zuogaiduoma town) of Hezuo in Gannan Tibetan autonomous prefecture.Thirty of child-bearing age,pregnant and breastfeeding women were selected,respectively,and 90 male adults aged 20-50 from these families(1 from each family) and 90 children aged 8-10 (30 people in each age group) from local schools were randomly sampled at the same time,and urinary iodine (UI) was measured randomly.Edible salt and main food samples were collected to test iodine content from the 10 families of the three types of women,respectively,and they were asked to recall its family intake of food species in the past 24 h excluding spices.The water iodine was determined using arseniccerium redox method (GB/T 5750.1-2006) ; UI with ammonium persulfate digestion-arsenic cerium catalytic spectrophotometric method (WS/T 107-2006) ; salt iodine used direct determination method(GB/T 13025.7-1999); and food iodine with alkali the gray arsenic cerium contact colorimetry.All these work were done in May,2011.Results The average of water iodine was (1.63 ± 0.14)μg/L in agricultural areas and (2.08 ±1.90)μg/Lin pastoral areas of the 10 water samples tested,respectively.The median urinary iodine(MUI) among women of pregnant,lactating and child-bearing age,male adults and children was 141.99,126.65,253.33,258.07,191.0μg/L,respectively,in agricultural areas and 137.26,97.36,126.16,159.48,285.07 μg/L,respectively,in pastoral areas.The difference of MUI in lactating,male adults and children between pastoral and agricultural areas was statistically significant.The proportion of UI ＜ 50 μg/L was less than 20％,and ＜ 100 μg/L was less than 50％ among all population except lactating woman and pregnant women in pastoral areas.The iodized salt coverage rate was 100％(30/30) in agricultural areas and 90％(27/30) in pastoral areas,and the salt iodine was (32.1 ± 7.8)mg/kg in agricultural areas and (32.3 ± 6.0)mg/kg in pastoral areas,respectively.The food structure in agricultural areas was mainly potato,naked oat fruit,cabbage and so on,the average dietary iodine content was 285.7 μg/kg,and in pastoral areas was mainly chow mein,wheat flour,ghee,yogurt,barley and so on,the average dietary iodine content was 51.1 μg/kg.Conclusions There is no iodine deficiency in general in the population in Tibetan areas with low water iodine.However,iodine nutrition of pregnant women can not be guaranteed.It is recommended that classified guidance measures be taken to ensure the sustainable elimination of iodine deficiency disorders in the Tibetan minority areas.

Objective: To observe the clinical efficacy of sinew-bone balancing manipulation in treating lumbar disc herniation (LDH) and offer clinical evidence to support the concept of paying equal attention to sinew and bone. Methods: Sixty LDH patients were randomized into an observation group and a control group, with 30 cases in each group. The observation group was treated with the sinew-bone balancing manipulation, and the control group received conventional Tuina (Chinese therapeutic massage) manipulation. The clinical efficacy and posterior muscle chain tone effect were compared between the two groups by observing the visual analog scale (VAS) score, Japanese Orthopaedic Association (JOA) score, and posture-associated indicators. Results: The total effective rate was 86.7％ in the observation group, higher than 76.7％ in the control group, but the between-group difference in efficacy was statistically insignificant (P>0.05). After treatment, the VAS and JOA scores, angle-dependent muscle tone indicator of the posterior muscle chain, and lumbar posture symmetry showed significant changes in both groups (P<0.05). The VAS and JOA scores, angle-dependent muscle tone indicator of the posterior muscle chain, and lumbar posture symmetry in the observation group were significantly different from those in the control group after treatment (P<0.05), but the between-group difference in the general posture symmetry was statistically insignificant (P>0.05). Conclusion: Both the sinew-bone balancing and conventional Tuina manipulations can reduce pain in LDH patients, improve lumbar function, and adjust the angle-dependent muscle tone coefficient of the posterior muscle chain and lumbar posture symmetry; except for the general posture symmetry of the posterior muscle chain, the sinew-bone balancing manipulation wins out over the conventional Tuina manipulation.

Objective To evaluate the individualized 3D) printed drilling guide we developed and used for placement of C1/2 pedicle screws in the clinical treatment of fracture and dislocation of the atlantoaxial joint.Methods From January 2014 to June 2016,we treated a total of 17 patients with fracture and dislocation of the atlantoaxial joint.All the cervical CT data of the patients were imported into the digital orthopaedic workstation for 3D reconstruction,data modeling and 3D printing to design and manufacture individualized atlantoaxial vertebral guide templates.Intraoperatively,C1 and C2 pedicle screws were placed under the guide of individualized 3D printed drilling template.Cervical short-segment fixation and fusion were conducted for the patients.Postoperatively,regular clinical and radiographic follow-ups were carried out.Results No serious complications like injury to spinal cord and vertebral artery happened due to failed placement of C1/2 pedicle screws.The operation time ranged from 136 to 222 min (average,168.0 rmin);the intraoperative blood loss ranged from 260 to 556 mL (average,356.0 mL).The 17 patients were followed up for 6 to 36 months (average,13.5 months).The patients obtained bony union after 4 to 6 months (average,4.8 months).At the final follow-up,according to ASIA92 scoring system,the average sensory score was improved significantly from preoperative 7.4 + 3.2 to postoperative 13.1 + 5.9,and the average motor score was improved significantly from preoperative 5.3 + 3.1 to postoperative 11.7 + 5.1 (P ＜ 0.05).No such complications as infection or implant failure occurred after operation.Conclusion Individualized 3D printed drilling guide for implantation of atlantoaxial pedicle screws can make the complicate and risky placement become accurate,safe and simple.

Objective To determine the proportion of CD4+ CD25+ regulatory T (Treg) cells,mRNA expression of the forkhead box protein 3 (Foxp3) gene,and DNA methylation status of the Foxp3 promoter in peripheral CD4+ T cells from patients with Henoch-Sch(o)nlein purpura.Methods Totally,20 inpatients with Henoch-Sch(o)nlein purpura and 20 healthy controls were enrolled from Department of Dermatology,the Second Xiangya Hospital of Central South University between 2015 and 2016,and there were no significant differences in the gender and age between the two groups (both P ＞ 0.05).CD4+ T cells were isolated from the peripheral blood samples of these subjects.Real-time fluorescence-based quantitative PCR was performed to detect the mRNA expression of the Foxp3 gene,flow cytometry to determine the proportion of CD4 + CD25+ Treg cells,and sodium bisulfite sequencing PCR (BSP) to determine the DNA methylation status of the Foxp3 promoter.Statistical analysis was carried out with SPSS16.0 software by using two-sample t test for the comparison between the two groups,and linear correlation analysis for evaluating the correlations of the DNA methylation status of the Foxp3 promoter with clinical severity scores and the proportion of CD4+CD25+ Treg cells.Results Compared with the healthy control group,the Henoch-Sch(o)nlein purpura group showed significantly decreased mRNA expression of the Foxp3 gene in CD4+ T cells (0.380 ± 0.226 vs.1,t =9.503,P ＜ 0.01),proportion of CD4+CD25+ Treg cells (1.668％ ± 0.959％ vs.2.741％ ± 1.131％,t =2.552,P ＜ 0.05),but significantly increased DNA methylation status of the Foxp3 promoter (0.712 ± 0.164 vs.0.453 ± 0.147,t =3.610,P ＜ 0.01).In the Henoch-Sch(o)nlein purpura group,the DNA methylation status of the Foxp3 promoter was negatively correlated with the percentage of CD4+CD25+ Treg cells (r =-0.490,P ＜ 0.05),but positively correlated with the clinical severity scores (r =0.486,P ＜ 0.05).The DNA methylation level of the Foxp3 promoter was significantly higher in the patients with renal impairment than in those without renal impairment (P ＜0.05).Conclusion The patients with Henoch-Sch(o)nlein purpura showed increased DNA methylation status of the Foxp3 promoter in CD4+ T cells,decreased mRNA expression of the Foxp3 gene and proportion of CD4+CD25+ Treg cells,which may be related to the occurrence of Henoch-Sch(o)nlein purpura,and affect disease development and prognosis.

Background Statistic data revealed that different retinopathy of pre-term infants have different susceptibility to retinopathy of prematurity (ROP),which may be associated with polymorphism of vascular endothelial growth factor(VEGF) gene.Objective This study was to determine the association of polymorphisms of VEGF gene with the risk for ROP.Methods This research was approved by Ethics Committee of Hunan Children's Hospital,and written informed consent was obtained from the parents of patients.A prospective case-controlled study was designed.Ninety-nine ROP patients in Hunan Children' s Hospital and 88 pre-termed children without ROP were included from January,2006 to December,2009.Thirty-nine patients who received retinal photocoagulation or cryotherapy were included as the treatment group,and 60 untreated but spontaneously regressed ROP patients as the non-treatment group.No significant differences were seen in demography between with the ROP group and the without ROP group,or between the treatment group and the non-treatment group (all P＞0.05).2 mL of peripheral blood was collected for the extraction of DNA.Gene polymorphisms of VEGF-A+405 and VEGF-A936 were detected using the pyrosequencing method.Results No significant difference was found in the frequencies of the VEGF-A+405 gene polymorphisms between the ROP group and without ROP group (P =0.071,OR =0.675,95 ％ CI =0.444-1.026).Also no significant difference was found in the frequencies of the VEGF-A936 gene polymorphisms between with the ROP group and without the ROP group (P =0.118,OR =0.768,95 ％ CI=2.823-4.614).However,the frequencies of the VEGF-A+405 gene polymorphisms were significantly higher in the ROP treatment group than the non-treatment group (P＜0.01,OR--0.857,95 ％ CI =5.239-14.024),and VEGF-A936 gene polymorphisms was also significantly higher in the ROP treatment group than the non-treatment group (P =0.000,OR =3.609,95 ％ CI =0.711-0.829).Conclusions There is no association between the VEGF-A+405/VEGF-A936 single nucleotide polymorphism with the risk of ROP,but polymorphisms of VEGF gene may be correlated with the prognosis of ROP.The carrier of VEGF-A +405 /VEGF-A936 allele is more susceptible to ROP progression.

OBJECTIVETo compare different expression profiles of all known chemokine receptors in human hepatocellular carcinoma (HCC) cell lines with different metastasis potentials.

METHODSEighteen pairs of chemokine receptor primers were designed using Premier software. Expression profiles of the 18 chemokine receptors on four HCC cell lines of lower to higher potentials of metastasis (SMMC-7721, MHCC97-L, MHCC97-H and HCCLM6) were analyzed by RT-PCR. Expression of CXCR4 was detected by RT-PCR.

RESULTSExpression profiles of chemokine receptors on four HCC cell lines with different metastatic potentials had significant differences (P < 0.01), in which CCR10, CXCR4 and CXCR6 expressions decreased gradually as the metastatic potential of the cell lines increased. The expressions of CCR3, CCR4, CCR10, CCR12 and XCR1 on HCCLM6 were significantly reduced compared with SMMC-7721 (P < 0.01), whereas the expressions of CXCR1 (P = 0.006) and CXCR5 (P = 0.003) exceeded that of SMMC-7721. Except for CXCR2, CXCR6 and XCR1, most of chemokine receptors on MHCC97-H were expressed differently compared with MHCC97-L (P < 0.05), in which expressions of CCR1 (P = 0.002), CCR2 (P = 0.004) and CCR5 (P = 0.046) exceeded MHCC97-L. CXCR4 was detected only on the positive controls and SMMC-7721 when the template of total RNA was reduced one-half in RT-PCR.

CONCLUSIONChemokine receptors are expressed very differently at mRNA level on HCC cell lines with different metastatic potentials. The different profiles of chemokine receptors in tumor microenvironment and the function of CXCR4 in HCC should be further studied. Our findings have important implications in understanding the relationship between chemokine receptors and the metastatic potential of HCC.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; metabolism ; pathology ; RNA, Messenger ; genetics ; Receptors, Chemokine ; metabolism

OBJECTIVETo analyze the value of ovarian reserve markers for predicting ovarian response in women undergoing in vitro fertilization-embryo transfer.

METHODSAccording to the ovarian response, 331 patients undergoing oocyte retrieval cycles were divided into of normal, poor, and high response groups. Serum anti-Mvllerian hormone (AMH) was determined using AMH ELISA kit on day 3 of the menstrual cycle, antral follicle count (AFC) was measured using vaginal ultrasound, and basal serum follicle-stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E(2)) levels were detected using chemiluminescence method.

RESULTSSerum AMH and FSH levels, FSH/LH ratio, AFC, and the patients age, but not the basal E(2) level (P>0.05), were correlated with the number of oocytes collected (×1000/ampules of Gn) (P<0.001). AFC and serum AMH were the strongest single predictors for low ovarian response, with the areas under curve (AUC) of 0.855 (0.787-0.924) and 0.832 (0.764-0.900) (P<0.05), and cutoff values of ≤9 and ≤1.88 ng/ml, respectively. AFC was the strongest single predictor for high ovarian response, with an AUC of 0.787 (0.728-0.847) and the cutoff value of ≥15. Logistic regression model found that the combination of AFC, serum AMH and FSH improved the predictive power for poor ovarian response, but not for high ovarian response.

CONCLUSIONAFC, serum AMH, FSH, FSH/LH, and age are all predictors of ovarian response, among which AFC is the strongest single predictor. A multivariable model can improve the predictive power for low ovarian response but not for high ovarian response.

Adult ; Age Factors ; Anti-Mullerian Hormone ; blood ; Embryo Transfer ; Estradiol ; blood ; Female ; Fertilization in Vitro ; Follicle Stimulating Hormone ; blood ; Humans ; Luteinizing Hormone ; blood ; Middle Aged ; Oocytes ; cytology ; Ovarian Follicle ; cytology ; metabolism ; Ovary ; cytology ; metabolism ; Ovulation Induction ; methods ; Young Adult