Pancreatic ß-Cell O-GlcNAc Transferase Overexpression Increases Susceptibility to Metabolic Stressors in Female Mice.

The nutrient-sensor O-GlcNAc transferase (Ogt), the sole enzyme that adds an O-GlcNAc-modification onto proteins, plays a critical role for pancreatic ß-cell survival and insulin secretion. We hypothesized that ß-cell Ogt overexpression would confer protection from ß-cell failure in response to metabolic stressors, such as high-fat diet (HFD) and streptozocin (STZ). Here, we generated a ß-cell-specific Ogt in overexpressing (ßOgtOE) mice, where a significant increase in Ogt protein level and O-GlcNAc-modification of proteins were observed in islets under a normal chow diet. We uncovered that ßOgtOE mice show normal peripheral insulin sensitivity and glucose tolerance with a regular chow diet. However, when challenged with an HFD, only female ßOgtOE (homozygous) Hz mice developed a mild glucose intolerance, despite increased insulin secretion and normal ß-cell mass. While female mice are normally resistant to low-dose STZ treatments, the ßOgtOE Hz mice developed hyperglycemia and glucose intolerance post-STZ treatment. Transcriptome analysis between islets with loss or gain of Ogt by RNA sequencing shows common altered pathways involving pro-survival Erk and Akt and inflammatory regulators IL1ß and NFkß. Together, these data show a possible gene dosage effect of Ogt and the importance O-GlcNAc cycling in ß-cell survival and function to regulate glucose homeostasis.

Characteristics of bone metabolism in postmenopausal female patients with different types of idiopathic benign paroxysmal positional vertigo: A single-centre retrospective study.

OBJECTIVE: The association between benign paroxysmal positional vertigo (BPPV) and impaired calcium metabolism has attracted widespread interest. Several studies have suggested that decreased bone mineral density (BMD) and serum 25-hydroxyvitamin D (25(OH)D) level are related to the occurrence and/or recurrence of BPPV; however, the characteristics of bone metabolism in patients with BPPV subtypes have not been fully investigated, and conclusions have been controversial. This study aimed to evaluate BMD and serum levels of 25(OH)D and bone turnover markers to clarify the characteristics of bone metabolism in patients with different types of BPPV. METHOD: We retrospectively analysed the data of new-onset idiopathic postmenopausal female patients with BPPV at our institution from January 2016 to January 2020. The patients' demographic data including age, medication history, concomitant diseases, onset time, clinical form, laboratory indicators, such as serum levels of 25(OH)D, bone formation markers, namely, amino-terminal propeptide of type I procollagen (PINP) and osteocalcin (OC), bone resorption marker, namely, ß-isomerized carboxy-terminal telopeptide of type I collagen (ß-CTX), and BMD were collected and analysed. RESULTS: This study included 201 consecutive postmenopausal female patients with BPPV. Among them, 138 were diagnosed with posterior semicircular canal BPPV, 42 were diagnosed with lateral semicircular canal canalolithiasis, and 21 were diagnosed with lateral semicircular canal cupulolithiasis. There were no significant differences in age distribution, body mass index, clinical history, levels of albumin, globulin, uric acid, creatinine, or blood urea nitrogen, lipid profiles (except high-density lipoprotein cholesterol) and routine blood parameters among these groups (P > 0.05). There were no significant differences in the mean T-score and BMD values of different sites or in the serum levels of 25(OH)D and bone turnover markers (PINP, OC and ß-CTX) among the subgroups (P > 0.05). The proportion of reduction in BMD (T-score < -1 SD) and decreased serum vitamin D level (< 20 ng/ml) were not significantly different between the subgroups (P > 0.05). CONCLUSION: There were no significant differences in bone metabolism in postmenopausal female patients with different types of idiopathic BPPV.

The immediate impact of the 2019 novel coronavirus (COVID-19) outbreak on subjective sleep status.

BACKGROUND: An outbreak of the 2019 novel coronavirus (COVID-19) has been ongoing in China since January 2020. The threat of infection affects the work and life of most of the population and may also damage sleep. This study aims to examine the subjective sleep status and mental health of the population during the peak of the COVID-19 epidemic. METHOD: The data were collected through an online questionnaire with a sample of 5461 individuals in China from February 5, 2020, to February 23, 2020. Participants were divided into four groups based on their degree of threat from COVID-19: Group 1 was most closely associated with COVID-19, including inpatients diagnosed with COVID-19, first-line hospital workers and first-line management staff; Group 2 included outpatients diagnosed with COVID-19 and patients who developed a fever and visited the hospital; Group 3 included people related to Group 1 or 2, such as their colleagues, relatives, friends and rescuers; and Group 4 was the farthest removed from contact with COVID-19, covering the general public affected by COVID-19 prevention strategies. The Insomnia Severity Index (ISI), Patient Health Questionnaire (PHQ-9), Generalized Anxiety Disorder Scale (GAD-7) and Acute Stress Disorder Scale (ASDS) were used. RESULTS: Threat degree of COVID-19 (groups) had significant correlations with insomnia, depression, anxiety, and stress (p < 0.05, p < 0.01). Age, gender, and area (Hubei province or other provinces) had significant correlations with insomnia (p < 0.01). A total of 1380 (24.46%) participants were suspected of having major depression based on the PHQ-9. Additionally, 1042 (18.47%) participants were suspected of having generalized anxiety disorder based on the GAD-7. A total of 892 (15.8%) of the participants had Acute Stress Disorder (ASD) according to the ASDS. The prevalence of clinical insomnia during the outbreak was 20.05% (1131) according to the ISI. The factors of satisfaction with the current sleep pattern and how perceptible the symptoms of the current sleep pattern are to other people (p < 0.05) and the middle (difficulty staying asleep) and terminal (waking up too early) (p < 0.01) factors of the ISI were significantly different across groups. A total of 1129 (20.01%) participants spent more than one hour awake in bed. CONCLUSION: The results indicated that insomnia is more severe in people who are female, young, living in the epicenter and experiencing a high degree of threat from COVID-19. As prevention and treatment efforts continue with regard to COVID-19, the general public has developed poor sleep hygiene habits, which deserve attention.

Sestd1 Encodes a Developmentally Dynamic Synapse Protein That Complexes With BCR Rac1-GAP to Regulate Forebrain Dendrite, Spine and Synapse Formation.

SEC14 and Spectrin domain-1 (Sestd1) is a synapse protein that exhibits a striking shift from the presynaptic to postsynaptic space as neurons mature postnatally in the mouse hippocampus. Hippocampal pyramidal neurons from mice with global genetic deletion of Sestd1 have reduced dendrite arbors, spines, and excitatory synapses. Electrophysiologically this correlates with cell-autonomous reductions in both AMPA- and NMDA-excitatory postsynaptic currents in individual hippocampal neurons from which Sestd1 has been deleted in vivo. These neurodevelopmental and functional deficits are associated with increased activation of the Rho family GTPases Rac1 and RhoA. Co-immunoprecipitation and mass spectrometry reveal that the Breakpoint Cluster Region protein, a Rho GTPase activating protein (GAP), forms complexes with Sestd1 in brain tissue. This complements earlier findings that Sestd1 can also partner with other Rho family GAPs and guanine nucleotide exchange factors. Our findings demonstrate that Sestd1 is a developmentally dynamic synaptic regulator of Rho GTPases that contributes to dendrite and excitatory synapse formation within differentiating pyramidal neurons of the forebrain.

Chemical composition, antioxidant and antimicrobial activity of the essential oil from the leaves of Macleaya cordata (Willd) R. Br.

The essential oil from the leaves of Macleaya cordata R.Br. obtained by hydrodistillation was analysed by gas chromatography/mass spectrometry. Sixty-eight compounds consisting of up to 92.53% of the essential oil were identified. Antioxidant activities of the essential oil were evaluated by using DPPH radical scavenging and ß-carotene-linoleic acid assays. The essential oil showed moderate antioxidant activity. In addition, the essential oil exhibited potential antimicrobial activity against all tested microorganisms, with diameters of inhibition zones ranging from 8.7 ± 0.5 to 17.2 ± 1.2 mm and minimum inhibitory concentration values from 125 to 500 µg/mL. We selected the most sensitive bacterium Staphylococcus aureus as model to observe of the action of essential oils of M. cordata on the membrane structure by scanning electron microscopy. The treated cell membranes were damaged severely. The results presented here indicate that the essential oil of M. cordata may be potential sources of antioxidant and antimicrobial agents in the future.

[Analysis of Prokaryotic Community Structure in River Waters of the Ningbo Sanjiang Mouth].

The prokaryotic community structure in river waters of the Ningbo Sanjiang Mouth was analyzed for the first time using 16S rRNA gene based-Illumina Miseq high-throughput sequencing. A total of 215 504 high-quality sequences were obtained, and the results of alpha-diversity analysis revealed that Yongjiang River Watershed (YRW) harbored high diversity and richness of prokaryotic communities. Taxonomic assignment analysis indicated that ß-Proteobacterium, Actinobacteria and Bacteroidetes dominated in the river water of YRW, and accounted for 78. 88% of the total prokaryotic communities. Hydrological condition may play an important role in influencing the composition and structure of YRW prokaryotic community. In addition, several kinds of sewer- and fecal-pollution indicator bacterial groups were observed in this area with the highest abundance of pollution indicator bacteria occurring in the water sample of Yuyao River, implying that the Yuyao River might have a high potential risk of sewer- and fecal-pollution. Moreover, a total of 76 species and 18 subspecies of potential pathogenic bacteria, which accounted for 2. 19% and 0. 40% of total sequences respectively, were identified using BLASTN analysis with a local pathogenic bacteria database. Overall, this study provided an important basic data for shedding light on the structure and ecological function of YRW prokaryotic community.

[Reductive Dechlorination of Trichloroethylene by Benzoate-Enriched Anaerobic Cultures].

Gas chromatography was used to monitor the reductive dechlorination of trichloroethylene (TCE) by anaerobic enrichment cultures with benzoate as the sole carbon source. The 454 pyrosequencing technique was used to investigate the microbial community and the real-time quantitative PCR was used to quantify the gene copies of Dehalococcoides spp. (DHC). The results showed that TCE was dechlorinated to vinyl chloride along with the formation of methane in 94 days. The anaerobic enrichment cultures exhibited a high diversity, which were classified into 16 phyla, 33 classes, 52 orders, 88 families and 129 genera, while 51.2% of them belonged to unclassified group, which inferred that there were a large portion of bacteria with unknown functional in this system. Degradation of TCE was accomplished by reductive dechlorinating and other functional populations, and the DHC which carried tceA gene could be the dominant reductive dechlorinating populations in the system.

Vitamin D3 and insulin combined treatment promotes titanium implant osseointegration in diabetes mellitus rats.

This study investigated the effect of insulin and vitamin D(3) (VD(3)) treatment on implant osseointegration in diabetic mellitus (DM) rats. DM was induced by administration of streptozotocin in rats, which received implants insertion in the femur. Then animals were subjected to different treatment and divided to the following group: control, diabetic, insulin-treated diabetic, VD(3)-treated diabetic, insulin and VD(3) combination-treated diabetic rats. The glucose levels and weight of rats were periodically evaluated, and serum 25(OH)D(3) levels in rats were measured at the end of the experiment. Animals were sacrificed at 12 weeks after surgery, the peri-implant trabecular microstructure, implant fixation and implant osseointegration were measured by microscopic computerized tomography (micro-CT) evaluation, push-out test and histomorphometric analysis. Diabetic rats displayed significantly higher blood glucose level, lower body weight, lower serum 25(OH)D(3) levels, and less implant osseointegration than controls. Insulin treatment showed restorative effect on body weight and serum 25(OH)D(3) levels of diabetic rats, but the blood glucose level in diabetic rats were still substantially higher compared to controls after 14 days therapy of insulin. Combined treatment restored hyperglycemia in diabetic rats to be normal, and reversed the impaired osseointegration capacity of implants, with the bone volume ratio and percent osseointegration increased by 1.37-fold and 1.6-fold in micro-CT evaluation, the maximal push-out force and ultimate shear strength by 1.3-fold and 2.1-fold in push-out test, and the bone-to-implant contact and bone area ratio increased by 2.57-fold and 1.44-fold in histomorphometric analysis. Monotreatment also enhanced implant fixation, but less. These results indicated that insulin and VD(3) combined treatment may be an effective approach to enhance implant fixation in diabetic rats, but whether the results could be extrapolated to human needs further study.

A retrospective comparison of the efficacy and safety in kidney transplant recipients with basiliximab and anti-thymocyte globulin.

BACKGROUND: Induction therapy are utilized to achieve an adequate immunosuppression at the time of transplantation. The use of basiliximab or anti-thymocyte globulin (ATG) for induction therapy has significantly reduced the incidence of acute rejection episodes post-transplantation. The purpose of this study was to compare the efficacy and safety of the basiliximab in patients with immuno-induction therapy after kidney transplantation with the ATG. METHODS: A retrospective analysis was carried out in kidney transplant recipients including 146 patients with the basiliximab and 116 cases with the ATG and the acute rejection, graft function, infective complications and 1-year and 5-year actuarial patient and graft survival after renal transplantation were compared between the two treatment groups. RESULTS: There were no statistically significant difference between groups regarding age, sex, cold ischemic time, warm ischemic time, human leukocyte antigen (HLA) matching type between the donor and recipient, lymphotoxin test and the use of immunosuppressive agents. There was no statistical significance regarding the incidence of the acute rejection (9.59% vs. 8.62%, P = 0.481) and delayed graft function (10.27% vs. 9.48%, P = 0.501) between groups. There were significantly lower lung infection incidence (5.48% vs. 12.93%, P = 0.029) in the basiliximab-treated group in comparison with the ATG-treated group. One-year patient and graft survival rates were 98%, 97% for the basiliximab-treated group, and 95%, 73% for the ATG-treated group, respectively. Five-year patient and graft survival rates were 92%, 86% for the basiliximab-treated group and 93%, 72% for the ATG-treated group, respectively. Log rank test showed statistically significant difference with P = 0.038 for patients and P = 0.033 for grafts, respectively. There were significantly lower the incidence of granulocytopenia (8.22% vs. 17.24%, P = 0.022) and thrombocytopenia (4.11% vs. 19.83%, P = 0.000) after transplantation in the basiliximab-treated group in comparison with the ATG-treated group. There was no statistical significance regarding the incidence of the heart dysfunction after transplantation between the two groups (6.16% vs. 6.90%, P = 0.502). CONCLUSION: The immuno-induction therapy with the basiliximab in kidney transplant recipients is efficient and safe with less complication compared with the ATG.

Forkhead box protein 3 mRNA expression in the peripheral blood of kidney-transplant recipients with acute rejection.

BACKGROUND: Regulatory T cells (Tregs) are immunologically and clinically interesting not least because of the important role they play in allograft rejection. Likewise, expression of the transcription factor forkhead box protein 3 (FOXP3), detected in transplant biopsies, is also of interest because of its role in the development of regulatory T cells. In this study, we investigated the relationship between FoxP3 mRNA expression and acute organ rejection in kidney-transplant recipients. METHODS: In this prospective study, FoxP3 mRNA expression levels in peripheral blood samples from 10 recipients of living relative-donor kidney transplants were measured before transplantation as well as at the 14th and 90th days post-transplantation. In addition, 46 first-time kidney-transplant recipients participated in a cross-sectional study, with 28 patients classified as having acute organ rejection; whilst the remaining 18 patients had functionally stable allografts. FoxP3 mRNA expression levels in peripheral blood samples were compared between these two different groups. RESULTS: Before transplantation mean FoxP3 mRNA levels vs. GADPH mRNA levels (lg(FoxP3 mRNA/GADPH mRNA)) in the 10 recipients were 1.11 ± 0.67. The mean FoxP3 mRNA expression levels measured at 14th and 90th days post-transplantation were significantly higher than before transplantation (1.69 ± 0.38, P = 0.03; 1.44 ± 0.21, P = 0.04, respectively). Additionally, the mean FoxP3 mRNA levels vs. GADPH mRNA expression levels (lg(FoxP3 mRNA/GADPH mRNA)) were significantly higher in recipients suffering acute rejection compared with those with stable allografts (1.77 ± 0.61 and 1.43 ± 0.27, respectively, P = 0.03). CONCLUSIONS: After kidney transplantation, FoxP3 mRNA levels were found to increase in the peripheral blood of all recipients. Considerably higher FoxP3 mRNA levels were observed in recipients suffering acute rejection. These results suggest that FoxP3 mRNA levels in peripheral blood samples can be used as a diagnostic tool for identifying acute rejection.

All Dact (Dapper/Frodo) scaffold proteins dimerize and exhibit conserved interactions with Vangl, Dvl, and serine/threonine kinases.

BACKGROUND: The Dact family of scaffold proteins was discovered by virtue of binding to Dvl proteins central to Wnt and Planar Cell Polarity (PCP) signaling. Subsequently Dact proteins have been linked to a growing list of potential partners implicated in ß-catenin-dependent and ß-catenin-independent forms of Wnt and other signaling. To clarify conserved and non-conserved roles for this protein family, we systematically compared molecular interactions of all three murine Dact paralogs by co-immunoprecipitation of proteins recombinantly expressed in cultured human embryonic kidney cells. RESULTS: Every Dact paralog readily formed complexes with the Vangl, Dvl, and CK1δ/ε proteins of species ranging from fruit flies to humans, as well as with PKA and PKC. Dact proteins also formed complexes with themselves and with each other; their conserved N-terminal leucine-zipper domains, which have no known binding partners, were necessary and sufficient for this interaction, suggesting that it reflects leucine-zipper-mediated homo- and hetero-dimerization. We also found weaker, though conserved, interactions of all three Dact paralogs with the catenin superfamily member p120ctn. Complex formation with other previously proposed partners including most other catenins, GSK3, LEF/TCF, HDAC1, and TGFß receptors was paralog-specific, comparatively weak, and/or more sensitive to empirical conditions. CONCLUSIONS: Combined with published functional evidence from targeted knock-out mice, these data support a conserved role for Dact proteins in kinase-regulated biochemistry involving Vangl and Dvl. This strongly suggests that a principal role for all Dact family members is in the PCP pathway or a molecularly related signaling cascade in vertebrates.

Factors affecting the long-term renal allograft survival.

BACKGROUND: In the past decades, the one-year graft survival of cadaveric renal allografts has been markedly improved, but their long-term survival has not kept pace. The attrition rate of renal allografts surviving after one year remains almost unchanged. The causes for late graft loss are multiple. The aim of this study was to analyze the predictive factors that impact long-term survival of grafts after kidney transplantation. METHODS: We retrospectively analyzed 524 kidney transplantation patients who were treated in our hospital between January 1991 and January 2000, including 254 patients who had lived more than 10 years with normal graft function (long survival group), and 270 cases whose renal graft had survived less than 10 years (control group). Specifically, we analyzed 10 factors that may potentially affect graft survival by both univariate and Logistic model multivariate analyses to pinpoint the independent risk factors. RESULTS: Univariate analyses showed that no significant differences existed in the age or gender of recipients, dialysis time, lymphotoxin levels, or cold ischemia time between the two groups. However, the ratio of delayed graft function and acute rejection, and the uric acid levels of patients in the long survival group were significantly lower than those in the control group (P < 0.01). Furthermore, we found that the concentration of cyclosporin A at one year after transplantation and the histocompatibility antigen match of donor-recipients for patients within the long survival group were significantly higher than those in the control group (P < 0.01). Furthermore, multivariate analyses showed that these four factors were independent risk factors that impact patient survival. CONCLUSIONS: The ratios of delayed graft function and acute rejection, the concentration of cyclosporin A at one year after transplantation, and serum uric acid levels are very important factors that affect the long-term survival of renal grafts.

Monitoring immune function after rapid corticosteroid reduction in kidney transplant recipients.

BACKGROUND: Long-term use of steroid with large dosage might cause many adverse effects in kidney transplant patients; reducing steroid dosage to a low level for maintenance is helpful in avoiding the side-effects, but meanwhile, acute rejection may rise to be a main concern. The present research monitored the immune function changes and the incidence of acute rejection and infection after rapid steroid reduction to investigate the safety of this strategy. METHODS: A prospective trial was conducted, using tacrolimus and mycophenolate mofetil as the basic immunosuppressive regimen, in addition to antibody induction with basiliximab. Corticosteroid dosage was rapidly reduced to 10 mg/d seven days post-transplantation in the experimental group, and the standard corticosteroid therapy was employed in the control group. Patient immunity was monitored by the Immune Cell Function Assay pre- and two weeks post-transplantation. The incidence of acute rejection and infection were compared between the experimental and control group. RESULTS: Comparison of intracellular adenosine triphosphate (iATP) values detected two weeks post-transplantation for the control group ((324 ± 45) ng/ml) and the experimental group ((345 ± 91) ng/ml) did not reveal a significant difference (P > 0.05). The incidence of acute rejection was analogous between groups (P > 0.05), while an increased incidence of infection was observed in the control group (53% (n = 16)) versus the experimental group (22% (n = 6), P < 0.05). Overall, recipients in the control group had longer and more recurrent infections than those in the experimental group (P < 0.05). Patients in the control group had a lower immune response ((235 ± 35) ng/ml) than those in the experimental group ((286 ± 16) ng/ml) when infection occurred (P < 0.05). CONCLUSION: Rapid reduction of steroid early after kidney transplantation does not lead to a significant rise in patient immunity. It is a safe and effective therapy for kidney transplant patients.

[Immunoadsorption in highly sensitized renal transplant recipients].

OBJECTIVE: to investigate the efficacy and safety of protein A immunoadsorption (IA) therapy in the prevention of highly sensitized renal transplant recipients with acute rejection. METHODS: a retrospective analysis of preoperative clinical data was carried out in 12 renal transplant patients in our hospital from March 2008 to October 2009 with a high level of panel reactive antibody (PRA). Serum PRA and IgG, IgM, IgA were detected at pre and post-treatment. The acute rejection responses and adverse effects were observed. RESULTS: at post-treatment, PRA of both Class I and II antibody [14% (4%, 27%) vs 86% (73%, 98%), 6% (0, 23%) vs 68% (34%, 88%), P < 0.01] and serum total IgG levels significantly decreased [(550 ± 341) g/L vs (1301 ± 393) g/L, P < 0.01]. IgA and IgM were also lower than those at pre-treatment [(144 ± 78) g/L vs (185 ± 93) g/L, (103 ± 48) g/L vs (131 ± 66) g/L, P < 0.01]. Five cases had acute rejection reversible by anti-thymocyte globulin (ATG) or combined IA (n = 2). Within 6 months, 1 cases developed Aspergillus pneumonia, 2 cases of Pneumocystis carinii pneumonia were cured. CONCLUSION: IA therapy can reduce antibody levels in highly sensitized renal transplant recipients. When supplemented with induction therapy, it can effectively prevent and reduce rejection after renal transplantation.

[Hurthle cell thyroid tumor: an analysis of 28 cases].

OBJECTIVE: To explore the clinical features and the combined treatment modality of Hurthle cell thyroid tumor (HCT). METHODS: Twenty-eight cases of HCT treated between 2001 and 2009 were analyzed retrospectively. RESULTS: The age of the patients ranged from 18 to 72 years (with a median of 46.5 years); 22 females and 6 males. The main symptoms were thyroid solitary node or mass (22 cases) and multiple nodule (6 cases), 2 cases with cervical lymph node metastasis. All of the patients underwent surgery, 11 cases with thyroid lobectomy, 11 cases with thyroid lobectomy plus isthmusectomy, 4 cases with subtotal thyroidectomy, and 2 cases with thyroid lobectomy plus isthmusectomy and combined with modified radical cervical lymph node dissection. Postoperative pathological examination showed that 22 cases were Hurthle cell adenomas and 6 cases were Hurthle cell carcinomas, 1 of them with cervical lymph node metastasis. Twenty-one patients with Hurthle cell adenomas were followed up for 6 months to 7.5 years (with a median of 45 months) and 6 patients with Hurthle cell carcinomas for 3 to 8 years (with a median of 54 months), with no recurrence and death case. CONCLUSIONS: HCT is a potential malignant neoplasm. There are some difficulties in the diagnosis of HCT by frozen section. Surgery is an effective treatment for HCT. L-Thyroxine can be used to inhibit TSH excretion.

Modulation of the beta-catenin signaling pathway by the dishevelled-associated protein Hipk1.

BACKGROUND: Wnts are evolutionarily conserved ligands that signal through beta-catenin-dependent and beta-catenin-independent pathways to regulate cell fate, proliferation, polarity, and movements during vertebrate development. Dishevelled (Dsh/Dvl) is a multi-domain scaffold protein required for virtually all known Wnt signaling activities, raising interest in the identification and functions of Dsh-associated proteins. METHODOLOGY: We conducted a yeast-2-hybrid screen using an N-terminal fragment of Dsh, resulting in isolation of the Xenopus laevis ortholog of Hipk1. Interaction between the Dsh and Hipk1 proteins was confirmed by co-immunoprecipitation assays and mass spectrometry, and further experiments suggest that Hipk1 also complexes with the transcription factor Tcf3. Supporting a nuclear function during X. laevis development, Myc-tagged Hipk1 localizes primarily to the nucleus in animal cap explants, and the endogenous transcript is strongly expressed during gastrula and neurula stages. Experimental manipulations of Hipk1 levels indicate that Hipk1 can repress Wnt/beta-catenin target gene activation, as demonstrated by beta-catenin reporter assays in human embryonic kidney cells and by indicators of dorsal specification in X. laevis embryos at the late blastula stage. In addition, a subset of Wnt-responsive genes subsequently requires Hipk1 for activation in the involuting mesoderm during gastrulation. Moreover, either over-expression or knock-down of Hipk1 leads to perturbed convergent extension cell movements involved in both gastrulation and neural tube closure. CONCLUSIONS: These results suggest that Hipk1 contributes in a complex fashion to Dsh-dependent signaling activities during early vertebrate development. This includes regulating the transcription of Wnt/beta-catenin target genes in the nucleus, possibly in both repressive and activating ways under changing developmental contexts. This regulation is required to modulate gene expression and cell movements that are essential for gastrulation.

Rapamycin instead of mycophenolate mofetil or azathioprine in treatment of post-renal transplantation urothelial carcinoma.

BACKGROUND: Malignant tumor is the most common complication occurred in transplant recipients. It is widely recognized that immunosuppressive treatments increase the risk of cancer in transplant recipients. The efficacy and safety of rapamycin (RPM) in combination with low-dose calcineurin inhibitor (CNI) in treating 15 renal allograft recipients which developed urothelial carcinoma were observed. METHODS: Immunosuppressive regimen in all recipients was altered with rapamycin to replace mycophenolate mofetil (MMF) or azathioprine (Aza). The initial loading dosage was 2 mg/d, and the next dosage was 1 mg/d. The dosage of rapamycin was carefully adjusted according to the blood drug level and concentration of the drug was maintained at 4 - 6 microg/L. In all the 15 patients, the calcineurin inhibitor was reduced down to one third of the original dosage after the rapamycin blood concentration became stable. Surgical treatment and intravesical instillation chemotherapy were carried out in all patients. Recurrence of the tumor was monitored throughout the study. Post-transplant renal function and side effects were also closely monitored. RESULTS: Among the 15 patients, 9 had no tumor recurrence in 2 years, 2 had tumor recurrences twice, and 4 had once. There was no acute rejection observed during RPM treatment. Post-transplant renal function in 11 patients was improved, with a decreased creatinine level. Hyperlipoidemia and thrombocytopenia were the most frequent adverse events which responded well to corresponding treatments. CONCLUSION: Among the renal allograft recipients with urothelial carcinoma, combination of rapamycin and low dose calcineurin inhibitor treatment is effective and safe.

[Recovery characteristic of donor's and receptor's renal function from age over 55 years living donors donate kidneys].

OBJECTIVE: To observe and research clinical characteristics and curative effect and safety of renal transplantation from living elderly donors donating kidneys. METHODS: Retrospective study on the 19 living kidney donors who were over 55 years old and on the renal transplantation operations completed by our center for the past few years. Among the 19 donors, with an average age of 58 years old. Their mean creatinine clearance was 81.7 +/- 2.2 ml/min. Among the 19 acceptors, with an average age of 34 years old. All kidney before the open circulation transplant performed routine 0 point puncture and histological examination. RESULTS: All donors smoothly spent their perioperative period without any surgical complications. All the donors keep their blood Cr in a normal range one week after the operation. There was no significant difference between posttransplantation one week and six month and one year in blood Cr and Ccr. Blood pressure and blood sugar didn't not have significant changes, urine protein(-). All receptors' renal functions recovered in early stages without DGF. 7 receptors who had Ccr lower than 80 ml/min had their blood Cr decreased slowly. Among the 19 kidneys donated, 3 donors' glomerulosclerosises were higher than 10 percent. CONCLUSIONS: The kidney source shortage is the main factor that restricts the development of the renal transplantation currently, undoubtedly, the application of elderly donors will expand the kidney source and save more uremic patients. Renal transplantation is safe and feasible with the help of living elderly donors.

[Experimental study on effect of Bailing Cpsule on dendritic cells in mice].

OBJECTIVE: Comparing with the classic immuno-supressors, to probe in the in vitro effect of Cordyceps Sinensis (CS) on the differentiation, maturation and function of dentritic cells (DCs), and further to explore its mechanism. METHODS: Mice myeloid DCs were cultured respectively with extraction of Bailing Capsule (CSE), a Chinese medical preparation made of CS, Rapamycin and Tacrolimus, and the effect of various drugs on phenotype of DCs was analyzed with flow cytometer. Then, using as the stimulator, the DCs cultured with different drugs were mixed and cultured with heterogenous lymphocytes for observing the stimulating capacity of DCs on cell proliferation. RESULTS: CSE showed no in vitro effect on phenotype markers and co-stimulation molecules of DCs, the difference between CSE and Tacrolimus was insignificant, while Rapamycin could reduce the two parameters. CSE showed a marked suppressive effect on DCs in stimulating leucocyte proliferation in a dose-dependent manner. CONCLUSION: CSE could affect the stimulating capacity of DCs on cell proliferation, which is probably by means of inhibiting the function of antigen presentating cells to block the presentation of extrinsic signal, and make the low immune response condition, thus to obtain the effect of immunosuppression.