Study of immunoregulatory effect of sirolimus on xenotransplantaion with arterial patch / 器官移植

Objective To investigate the immunoregulatory effect of sirolimus on the xenotransplantation with arterial patch. Methods The xenotransplantation of arterial grafts was taken from the wild-type Bama pigs to cynomolgus monkeys. The peripheral blood mononuclear cells of recipient monkeys at 14 days after xenotransplantation (POD14) were selected as subjects. Dimethyl sulphoxide (DMSO) was used in the control group (volume ratio of 1:1 000) and sirolimus was administered in the sirolimus experimental group (final concentration of 0.1 μmol/L and 0.5 μmol/L). The cells were cultured for 1.0 and 5.5 d, respectively. The activity of POD14 cells was evaluated. The DMSO control and sirolimus experimental groups (final concentration of 0.1 μmol/L) were established and cultured for 5.5 d. The quantity of T and B cells in POD14 cells was counted and the expression levels of cytokines and messenger RNA (mRNA) were quantitatively measured. Results Compared with the DMSO control group, the activity of POD14 cells was significantly decreased after sirolimus treatment at a final concentration of 0.1 and 0.5 μmol/L for 1.0 d (P<0.01-0.001). After sirolimus treatment at a final concentration of 0.1 and 0.5 μmol/L for 5.5 d, the activity of POD14 cells was significantly decreased (both P<0.001). Compared with the DMSO control group, the quantity of CD3+CD4+T cells and CD3+CD8+T cells in POD14 cells was significantly reduced after sirolimus treatment at a final concentration 0.1 μmol/L (P<0.05-0.01), whereas the quantity of CD3-CD20+B cells was considerably elevated (P<0.01). Compared with DMSO control group, the levels of interferon(IFN)-γ, interleukin(IL)-2, IL-4, IL-5 and IL-6 in the sirolimus experimental group were significantly down-regulated (P<0.05-0.001). The expression levels of IFN-γ, tumor necrosis factor(TNF)-α, IL-2, IL-4, IL-5 and IL-6 mRNA were significantly down-regulated (P<0.05-0.001). Conclusions Sirolimus inhibits the proliferation of POD14 cells in the recipient monkeys after xenotransplantation with arterial patch. The underlying mechanism is that the sirolimus can reduce the quantity of T cells and suppress the expression and secretion of immune rejection-related cytokines.

Analysis of TCIRG1 gene mutation in a Chinese family affected with infantile malignant osteopetrosis / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To detect potential mutation of the TCIRG1 gene in a boy with infantile malignant osteopetrosis.</p><p><b>METHODS</b>Target sequence capture and next-generation sequencing were applied for the proband and his parents to identify the causative mutation, and Sanger sequencing was used to verify the suspected mutation.</p><p><b>RESULTS</b>The proband manifested at 4 months of age with symptoms including anemia, thrombocytopenia, hepatosplenomegaly, and cephalus quadratus. X-ray revealed generalized increased bone density. A novel compound heterozygous mutation, c.796G to T (p.E266X) and c.1372G to A (p.G458S), were identified in the boy. His father and grandmother also carried the c.796G to T (p.E266X) mutation, and his mother carried the c.1372G to A (p.G458S) mutation. Neither mutation was found in the PubMed and ClinVar databases.</p><p><b>CONCLUSION</b>The novel compound heterozygous mutation c.796G to T (p.E266X) and c.1372G to A (p.G458S) probably underlies the disease in the proband. Above results may enrich the mutation spectrum of the TCIRG1 gene and provide new evidence for the molecular basis of infantile malignant osteopetrosis.</p>

Effect and mechanism of SS31 peptide on autophagy after spinal cord injury in rats / 中华创伤杂志

Objective To explore the eftect of SS31 peptide on autophagy after spinal cord injury (SCI) and possible mechanism.Methods Allen nethod was used to construct the spinal cord injury model in rats.Sprague-Dawley rats were randomly divided into sham surgery group (sham group),SCI group and SS31 peptide group,with 30 rats in each group.The sham group only received laminectomy.The rats in SCI group were sustained SCI and were given no intervention.The rats in SS31 group received SS31 peptide injection after SCI.Scores of Basso Beattie Bresnahan (BBB) motor functions were assessed at 6 h,1,3,7 and 14 d after the injury.The changes in related proteins of Beclin-1 and LC3-Ⅱ were also detected.Results Scores of BBB scale at 6 h and at days 1,3,7 and 14 after injury in SCI group (0,1.7 ±0.4,3.5 ±0.6,6.1 ±0.7,10.1 ±0.6) and SS31 peptide group (0,2.5 ±0.7,4.1 ±0.7,9.3 ±0.6,13.4 ±0.6) were lower than that in sham group (21 at all time points) (P ＜0.05).Scores of BBB scale at days 7 and 14 after injury in SS31 peptide group (9.3 ±0.6,13.4 ±0.6) was higher than that in SCI group (6.1 ± 0.7,10.1 ± 0.6) (P ＜ 0.05).There was no significant difference upon scores of BBB scale of SS31 peptide group at 6 h and at days 1 and 3 after injury (0,2.5 ±0.7,4.1 ± 0.7),compared with SCI group (0,1.7 ± 0.4,3.5 ± 0.6) (P ＞ 0.05).Compared with sham group,the expression of Beclin-1 in SCI group and SS31 peptide group was increased,reached a peak at day 3 (1.478 ± 0.030,1.841 ± 0.051),remained high level at day 7 (1.302 ± 0.049,1.551 ± 0.032) and showed high expression at day 14 (1.252 ±0.048,1.471 ± 0.062) (P ＜ 0.05).Compared with sham group,the expression of LC3-Ⅱ in SCI group and SS31 peptide group also increased,reached a peak at day 3 (0.348 ± 0.028,0.655 ± 0.052),remained high level at day 7 (0.301 ± 0.053,0.432 ± 0.052) and also showed high expression at day 14 (0.268 ± 0.049,0.371 ± 0.052) (P ＜ 0.05).The expressions of Beclin-1 and LC3-Ⅱ in SS31 peptide group at day 3 after injury were 1.841 ± 0.051 and 0.455 ± 0.052,higher than that in SCI group (1.478 ± 0.030,0.348 ± 0.028) (P ＜ 0.05).In SS31 peptide group at 6 h and days 1,7 and 14 after injury,the expressions of Beclin-1 (0.582 ± 0.028,0.723 ±0.049,1.551 ±0.032,1.471 ±0.062) and LC3-Ⅱ (0.172 ±0.031,0.256 ±0.051,0.432 ± 0.052,0.371 ± 0.052) had no significant difference in comparison with corresponding expressions of Beclin-1 (0.584 ±0.021,0.642 ±0.051,1.302 ±0.049,1.252 ±0.048) and LC3-Ⅱ (0.156 ± 0.019,0.184±0.050,0.301 ±0.053,0.268 ±0.049) in SCI group (P＞0.05).Conclusion SS31 peptide can improve motor function and enhance the autophagy of nerve cells after SCI in rats,which may be one of the mechanisms for SS31 peptide treating spinal cord injury.

Clinical application of MRCP for the diagnosis of the 60 cases of biliary pancreatitis / 重庆医学

Objective To explore the clinic value of magnetic resonance cholangiopancreatography(MRCP)for the diagnosis of biliary pancreatitis(BP).Methods MRCP manifestation of 60 cases of BP proved by clinic records was analyzed retrospectively. Results Of all the 60 cases,41 were found pancreatitis combined with gall bladder stones or biliary cystitis,19 combined with stones in common biliary duct.Dilation of biliary duct was found in 23 cases,dilation of pancreatic duct in 9 cases,dilation of both in 5 cases.Conclusion MRCP can clearly display the signs of biliary pancreatitis,and have high clinic value for its diagnosis.

Isolation methods and biological characteristics of bone marrow mesenchymal stem cells / 中国组织工程研究

BACKGROUND:Bone marrow mesenchymal stem cells(BMSCs)have the properties involving high proliferation capability,widely distribution,functional tissue repair after injury,as well as immune modulation,by which bring us extensive therapeutic possibilities.There are plenty of methods for isolation of BMSCs,yet,BMSCs exhibit discrepancies in varied growth stage and culture conditions.Up to now,there has been no agreement about the identification methods for cultured BMSCs.OBJECTIVE:To review the isolation methods and biological characteristics of BMSCs,and to compare the differential expression of BMSCs between in serum and serum-free medium,prior to and after proliferation,as well as before and after induction.METHODS:A computer-based online search was performed using key words of "bone marrow mesenchymal stem cells,isolation,culture,induce,marker,and characterization" to find documents published in the database of CNKI (http://dlib.cnki.net/kns50/)or Pubmed(http://www.ncbi.nlm.nih.gov/PubMed)from January 2003 to June 2009.The languages were limited Chinese and English.A total of 237 literatures were searched by the computer.RESULTS AND CONCLUSION:The positive rates of CD44 and CD34 of BMSCs isolated by the whole bone marrow culture were smaller than that of the density gradient centrifugation.However,BMSCs isolated by the whole bone marrow culture were superior to those isolated by the density gradient centrifugation in cell viability,proliferation rate,confluence time,as well as generation time.Other methods for BMSCs isolation had drawbacks of large cost and high requirement of experimental equipments.Following conditions were used to identify BMSCs:cell adherence,cell surface molecule labeling,strong self-proliferation ability,as well as potentials multi-directional differentiation.BMSCs exhibit differential expression between in serum and serum-free medium,prior to and after proliferation,as well as before and after induction.

The relationship between insulin resistance and left ventricular hypertrophy in patients with early chronic kidney disease / 中华内科杂志

Objective To analyze the relationship between insulin resistance (IR) and left ventricular hypertrophy ( LVH ) in patients with early chronic kidney disease ( CKD ). Methods Homeostatic model method was used for detecting insulin resistance index (Homa-IR) in 108 patients with early CKD and 25 normal healthy cases, and the other clinical data such as Hb, creatinine clearance rate (Ccr) , parathyroid hormone (PTH) , ambulatory blood pressure monitoring ( ABPM ) data, including day average systolic blood pressure (dSBP), day average diastolic blood pressure (dDBP), night average systolic blood pressure ( nSBP) , night average diastolic blood pressure ( nDBP) , 24-hour mean systolic blood pressure (mSBP), 24-hour mean diastolic blood pressure (mDBP), decline in the percentage of diastolic blood pressure at night (nDPD) and decline in the percentage of systolic blood pressure at night (nDPS) were also measured. Echocardiography was used for measuring LVH relevant data, and left ventricular mass index (LVMI) was calculated. Then, the relationship between LVH and IR and other clinical data were analyzed retrospectively. Results IR existed in early CKD patients. With the decline of Ccr, both the value of Homa-IR and the incidence of IR increased significantly (both P < 0. 05). LVH existed in early CKD patients and with the decline of Ccr, both LVMI value and the incidence of LVH increased significantly (both P <0. 01). Compared with the non-IR group, the IR group had higher LVMI value (P<0. 05) and higher incidence of LVH (P<0. 01). Compared with the non-LVH group, the LVH group had higher levels of Fins, 2hPG, Homa-IR (all P <0. 05), and higher incidence of IR (P <0. 01). The LVH group had significant lower levels of Ccr, Hb and nDPD (all P <0. 05) , higher levels of dSBP, dDBP, nSBP, nDBP, mSBP, mDBP and PTH (all P<0.05) than the non-LVH group. LVMI had significant positive correlations with 2hPG, Fins, Homa-IR, dSBP, nSBP, mSBP and PTH (r = 0.255, 0.373, 0.376,0.222,0.199,0.225,0.221,0.246, respectively; all P<0.05), but significant negative correlations with Hb and Ccr (r = -0.588,-0.313, respectively; both P<0.01). Multi-factor regression analysis showed that Hb, Homa-IR, and Ccr entered the regression equation (y = 167. 106 - 0. 755x_1 + 0. 250x_2 +0.322x_3, y = LVMI; 167. 106 = constant, t = 12. 138, P =0.000; x_1 =Hb, t= -6.800, P = 0. 000; x_2 = Homa-IR, t = 3. 229, P = 0. 002; x_3 = Ccr, t = 2. 898, P = 0. 005). Conclusion IR existed in early CKD patients and become more severe with the decline of renal function. IR had a significant correlation with LVH, and it may be an important risk factor for the development of LVH. Besides, both anemia and decline of renal function are also associated with LVH.

Analysis of the relative etiology of non-obstructive azoospermia / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the levels and changes of sexual hormone in serum and seminal plasma, and the epidermal growth factor (EGF), alpha-glucosidase, and fructose in the seminal plasma of non-obstructive azoospermic patients, and to assess the significance of the analysis of their chromosomes.</p><p><b>METHODS</b>Thirty-six non-obstructive azoospermic males and 26 normal males were chosen. The results in reproductive endocrine and genetics obtained by radioimmunoassay, colorimetry, and chromosomes G and C banding assay were compared and analyzed to explore into the causes of infertility.</p><p><b>RESULTS</b>The levels of FSH, LH, PRL and E2 in the serum of the non-obstructive azoospermic patients were higher than those of the control group, but the T level had no significant difference between the two groups. There were no obvious differences in the sexual hormones except in FSH of the seminal plasma. There were 2 cases of abnormal sex chromosomes, 2 cases of big Y chromosomes and 7 cases of small Y chromosomes in the non-obstructive azoospermic patients. No correlation was found between EGF and fructose in the seminal plasma.</p><p><b>CONCLUSION</b>The level of sexual hormone in serum, alpha-glucosidase in seminal plasma and abnormal chromosomes might be closely related to non-obstructive azoospermia.</p>

EFFECTS OF CROCODILE BONE GELATIN ON STRESS REACTION AND BLOOD-FORMING FUNCTION IN MICE / 中国海洋药物

To investigate the effects of Crocodile Bone Gelatin(CBG) on stress reaction and blood forming function in mice. Results showed CBG increased staying power for high temperature and low temperature, but didn′t increase anti exhaustion ability and hypoxia tolerance of mice. CBG increased platelet count, didn′t increase white blood cell count on Cyclophosphamidum causing blood forming functional obstruction.

Protective Effects of Luotai Injection Powder on Cerebral Ischemia / 中成药

Objective:To study the protecting effects of Luotai Injection Powder on cerebral ischemia.Methods: The protecting effects of Luotai Injection Powder on cerebral ischemia were studied on animal models of cerebral ischemia induced by ligaturing bilateral common carotid artery and decressing blood pressure. Results:Luotai Injection Powder (at doses of iv. 60mg/kg and 120mg/kg) significantly decreased the water content, capillary permeability and ischemia injury in brain of rats. In addition, it significantly increased the tolerance in mice at a dose of iv. 120mg/kg. Conclusions: Luotai Injction Powder has a protective effect on acute cerebral ischemia