Sedum sarmentosun bunge extraction ameliorated severe acute pancreatitis-induced lung injury: an experimental research / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the effect of Sedum sarmentosum Bunge Extract (SSBE) on severe acute pancreatitis (SAP) induced acute lung injury (ALI) model rats and their excessive inflammatory reactions.</p><p><b>METHODS</b>Forty-two healthy adult male Sprague-Dawley (SD) rats were randomly divided into 3 groups, the sham-operated control group (C), the SAP group (SAP), and the SSBE treated group (SSBE), 14 in each group. SAP induced ALl rat model was induced by retrograde injection of 5% sodium taurocholate (1 mL/kg) into the pancreatic duct. SSBE (100 m/kg) was administrated subcutaneously after the establishment of the SAP model. Equal dose of SSBE was injected again 12 h later. Equal volume of normal saline was administrated in the same way for rats in the C group and the SAP group. Rats were sacrificed after successful modeling and samples taken at 12 and 24 h. Pathological changes in the pancreas and the lung tissue were observed under light microscope. The ascites, serum amylase (AMS), wet/dry proportion (W/D) of the lung tissue, activities of myeloperoxidase (MPO), interleukin-1 (IL-1), IL-6, and tumor necrosis factor-alpha (TNF-alpha) were also measured.</p><p><b>RESULTS</b>Ascites and serum AMS activities significantly increased; MPO, IL-1, IL-6, TNF-alpha contents, and W/D ratio also significantly increased in the SAP group, when compared with the C group (P<0.05). Compared with the SAP group, those parameters were all attenuated in the SSBE group at 12 and 24 h (P<0.05, P<0.01). Pathological changes in the pancreas and the lung tissue were alleviated in the SSBE group under light microscope. The injury degree ranged between that of the C group and the SAP group.</p><p><b>CONCLUSION</b>SSBE could relieve the ALl in SAP model rats, which could be achieved through alleviating inflammation responses of SAP rats.</p>

Application of HLAMatchmaker analysis eplets mismatch of renal transplant matching / 中国应用生理学杂志

<p><b>OBJECTIVE</b>Eplets mismatch based on HLAMatchmaker software evaluates the clinical application of kidney transplantation.</p><p><b>METHODS</b>In 239 cases of renal transplant,merits of methods of the traditional HLA six antigen matcheing criteria, cross reaction groups standard and Eplets mismatch based on HLAMatchmaker standard were compared respectively.</p><p><b>RESULTS</b>The number of mismatchs with three methods in 239 cases, were grouped according to low-high mismatchs. The results revealed that HLAMatchmaker algorithm could significantly increase the number of low mismatchs group 54 (22.6%), compared with the HIA group 19(7.9%) and CREGs group 32 (13.4%). The comparison was discovered statistical significance among the three groups (P<0.001), so the comparison between each group was.</p><p><b>CONCLUSION</b>HLAMachmaker of donor-recipients matching, is a more efficient, time-saving and high sensitivity matching solution to allograft renal transplantation.</p>

The application of genotyping in the complicated ABO blood group type / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the application of ABO blood group genotyping in complicated ABO blood group type.</p><p><b>METHODS</b>Ten specimens of complicated ABO blood group were genotyped by sequence specific primer PCR (PCR -SSP), and confirmed by DNA sequencing and alignment. Six hundred and ten blood samples typed by ABO immunoassay were as control of genotyping.</p><p><b>RESULTS</b>Ten cases of complicated blood type were identified by high resolution PCR- SSP as rare ABO blood groups: cis-AB01 (3 cases), B(A)04 (2 cases), cisAB02, B(A)02, Bel03, Bw12 and Ael05, confirmed by DNA sequencing. Genotyping and serotype detected 610 cases ABO blood group were coincident, and the frequency of A, B, AB and O were as 28.69%, 27.54%, 8.2% and 35.57% respectively. According to the genotypes, the highest frequency subgroup was O1 (32.87%), the lowest was A2 (0.66%).</p><p><b>CONCLUSION</b>PCR -SSP could type the ABO blood group accurately, but also the sub-group of blood type. However, special designed high resolution PCR -SSP or DNA sequencing is needed to identify the complicated blood groups.</p>

The role of adenosine deaminase in the electroacupuncture preconditioning induced rapid tolerance to focal cerebral ischemia / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the electroacupuncture (EA) pretreatment at Baihui (GV20) on the concentration of adenosine deaminase (ADA) and adenosine, and to evaluate its effects on the neurologic function score and the infarction volume after middle cerebral artery occlusion (MCAO) ischemia/reperfusion (I/R), thus exploring its mechanisms for relieving the ischemia/reperfusion injury.</p><p><b>METHODS</b>Totally 54 male SD rats were randomly divided into 3 groups, the sham-EA group, the EA group, and the control group, 18 in each group. Rats in the control group were not intervened after anesthesia. Rats in the EA group were needled at Baihui (GV20) for 30 min. Rats in the sham-EA group received the same procedure as those performed in the EA group without electricity connected. The changes of adenosine and ADA contents were detected at 30, 60, and 120 min after EA respectively. The I/R model was established. Totally 48 male SD rats were randomly divided into 6 groups, i.e., the model group (Group A), the EA group (Group B), the EA +8-Cyclopentyl-1,3-dipropylxanthine (DPCPX) group (Group C), the EA + DMSO group (Group D), the Deoxycoformycin (Deo) group (Group E), and the normal saline group (Group F). Rats in Group B, C, and D received EA for 30 min before modeling. Rats in Group C and D were peritoneally injected with DPCPX (1 mg/kg) and DMSO (1 mL/kg) at 30 min before EA. The neurologic function score was evaluated and the infarct volumes were detected after 24-h reperfusion.</p><p><b>RESULTS</b>Compared with the sham-EA group, there was no statistical difference in the contents of the adenosine or ADA in the control group at each time point (P > 0.05). Compared with the control group at the same time point, the content of ADA significantly decreased at 60 min in the EA group [(315.0 +/- 22.9 U/L), P < 0.05], and restored to the normal level at 120 min after EA. The content of adenosine increased in the EA group at 120 min [(20.4 +/- 2.2) ng/microL, P < 0.05]. Compared with the model group, the neurologic function score decreased (P < 0.05) and the infarct volumes were obviously reduced (P < 0.01) in Group B, D and E. There was no statistical difference in the neurologic function score or the infarct volumes in other groups, when compared with the model group (P > 0.05)</p><p><b>CONCLUSION</b>EA at Baihui (GV20) showed protective effects on the cerebral I/R rats, which might be achieved through lowering the ADA concentration and elevating the adenosine content, and further activating adenosine A1 receptor.</p>

Analysis of a hereditary protein C deficient consanguineous pedigree caused by Phe139Val homozygous mutation / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyze genetic mutation and explore its molecular pathogenesis for an hereditary protein C (PC) deficient consanguineous pedigree.</p><p><b>METHODS</b>The pedigree included three generations and contained eight members. PC activity (PC:A), PC antigen (PC:Ag) and other coagulant parameters were detected for all family members. Protein C gene (PROC) include all the exons and intron exon boundaries were amplified by PCR for the proband, then analyzed by direct sequencing. Mutation sites were detected for the other family members.</p><p><b>RESULTS</b>The PC:A and PC:Ag in the proband plasma were 20% (normal range 70% -140%) and 13.2% (normal range 70%-130%). A homozygous missense mutation g.6128T>G in exon 7 resulting in Phe139Val was identified in the proband. The PC:A and PC:Ag in her younger brother were 31% and 18.90%, Phe139Val homozygous was also found. The left family members were heterozygous for Phe139Val.</p><p><b>CONCLUSION</b>Phe139Val homozygous missense mutation in exon 7 of PROC caused serious hereditary protein C deficiency. We speculated that homozygous mutation might be resulted from this consanguineous marriage.</p>

Study on KIR gene polymorphisms in 416 renal transplantation recipients from southern Zhejiang / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate polymorphisms of killer cell immunoglobulin-like receptor gene (KIR) in renal transplant recipients from southern Zhejiang.</p><p><b>METHODS</b>KIR genotypes were analyzed by PCR-SSP in 416 renal transplant recipients, and the genotype frequencies were compared with populations from Eastern China and worldwide.</p><p><b>RESULTS</b>All 16 known KIR genes were detected in the renal transplant recipients, and KIR2DL4, 3DL2-3, 3PD1 were found in all. As a pseudogene, 2DP1 has a high genotype frequency (99%). The frequencies of KIR2DL1, 2DL3, 3DL1, 2DS4 have ranged from 92.1% to 98.8%. Compared with 11 groups in Eastern China and other countries, the KIR2DL2 phenotype frequency was higher (34.6%) than those of Shanghai, Zhejiang and Jiangsu populations (P<0.05). Among 41 genotypes, three have not been reported previously. The most common genotype was AA1, with a frequency of 43.51%, which was significantly lower than those of Jiangsu and Northern Zhejiang.</p><p><b>CONCLUSION</b>Renal transplant recipients from southern Zhejiang share similar features with Eastern China Han population with regard to KIR polymorphisms, but also have unique frequencies for KIR genotypes.</p>

Effects of ginsenoside Rb1 on the mRNA expression of tyrosine kinase B in the hippocampus of acute immobilization stress rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effects of acute immobilization stress on the mRNA expression of tyrosine kinase B (TrkB) in rats' hippocampus.</p><p><b>METHODS</b>Eighteen SD rats were randomly divided into three groups, i.e., the normal control group, the model group, and the medication group, 6 in each group. The acute immobilization stress model was prepared in the model group using acute immobilization for 2 h. Ginsenoside Rb1 (40 mg/kg) was peritoneally injected to rats in the medication group 30 min before modeling, with the same procedure as those for rats in the model group. No treatment was performed to rats in the normal control group. The plasma adrenocorticotropic hormone (ACTH) and corticosterone (CORT) contents were detected using ELISA. The mRNA expression of TrkB in the rats' hippocampus was detected using real-time fluorescence quantitative RT-PCR.</p><p><b>RESULTS</b>Before modeling there was no statistical difference of plasma CORT or ACTH concentrations among three groups (P >0.05). The plasma CORT and ACTH concentrations increased in the model group and the medication group more significantly after modeling than before modeling, showing statistical difference (P <0.05). Besides, they were obviously higher in the model group than in the normal control group (P <0.05). They were obviously higher in the medication group than in the model control group (P <0.05). Compared with the normal control group, the mRNA expression of TrkB significantly decreased in the model group (87.73 +/- 7.62 vs 50.65 +/- 5.19, P < 0.05), showing statistical difference. The mRNA expression of TrkB was significantly higher in the medication group (78.91 +/- 18.07) than in the model group, showing statistical difference (P <0.05).</p><p><b>CONCLUSION</b>Pretreatment by ginsenoside Rb1 could increase the plasma CORT and ACTH concentrations, maintain the mRNA expression of TrkB, thus relieving injury induced by acute immobilization stress.</p>

Establishment of a real-time PCR assay for simultaneously detecting human BKV and CMV DNA and its application in renal transplantation recipients / 病毒学报

To establish a fluorescent quantitative PCR method (FQ-PCR) with TaqMan probe for simultaneous detection of polyomavirus (BKV) and cytomegalovirus (CMV) and to evaluate its clinical application in the renal transplantation recipients. The conservative sequences of BKV and CMV were targeted and amplified by nested PCR technique. The PCR products were cloned into the plasmids pcDNA3. 1(+). The recombinant plasmid containing target sequences of BKV and CMV were constructed as external standards. The TaqMan-based assay was optimized. For evaluating the assay, the sensitivity was determinated by diluted standard (5 X 103-10icopies/mL), and the specificity was verified by negative control and positive control, and the precision was assessed by intra-assay coefficient of variation (ICV) through detecting standard repeatedly (20 times). A total of 480 blood samples of renal transplantation recipients were used to detect BKV and CMV DNA simultaneously with FQ-PCR, and the concentrations of FK506 were measured by ELISA. The association of DNA copy and concentrations of FK506 was analyzed. The cloned target BKV and CMV DNA was confirmed by sequencing and analysis. The sensitivity of the FQ-PCR assay reached 5 X 103 copies/ml in detecting BKV or CMV DNA. Control DNA verified the assay specifically detecting target DNA. The precision of the assay to quantif target DNA copies was acceptable (Intra-assay CV was 3.44% for BKV and 2.23% for CMV; Inter-assay CV was 4. 98% for BKV and 3.76% for CMV;). Of 480 samples, 130 samples (27. 08%) were CMV DNA positive, significantly higher than the BKV DNA positive (13.33%, 64/480, P<0.05). The positive BKV or CMV DNA was found to be associated with high concentrations of FK506 (P<0. 05). In conclusion, the developed real-time PCR assay for detecting both CMV and BKV DNA simultaneously was s high sensitive, precise and time-effectiveand could be applied in the monitoring of the CMV and BKV infection in the renal transplantation recipients.

Analysis of an hereditary protein C deficiency pedigree with compound heterozygous gene mutations / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze genetic mutations and explore its molecular pathogenesis for an hereditary protein C (PC) deficiency pedigree.</p><p><b>METHODS</b>The pedigree has included 15 individuals from 4 generations. Plasma levels of PC activity (PC:A), PC antigen (PC:Ag) and other coagulant parameters were determined for members of the family. The 9 exons and intron-exon boundaries of protein C gene (PROC) of the proband were amplified with PCR and analyzed with direct sequencing. Detected mutations were confirmed with reverse sequencing. Corresponding PCR fragments from the family members were also directly sequenced.</p><p><b>RESULTS</b>Plasma PC:A and PC:Ag for the proband was 26% and 18.60%, respectively, both being lower than normal references. Seven members from the pedigree also had lower PC:A, six had lower PC:Ag. A compound heterozygous missense mutation, including a T to G transition at position 6128 of exon 7, which results in Phe139Val, and a G to C transition at position 8478 in exon 9, which results in Asp255His, were identified in the proband. The paternal grandma, father and two aunts were heterozygous for g.6128 T to G, whilst the mother, the second uncle, sister and son were heterozygous for g.8478 G to C. There were lower PC:A in family members with g.8478 G to C.</p><p><b>CONCLUSION</b>The proband had inherited two independent mutations of the PROC gene including g.6128 T to G in exon 7 and g.8478 G to C in exon 9 from her father and mother, respectively. The resulting compound heterozygous mutation has caused a serious hereditary protein C deficiency.</p>

Protective effect of sirolimus against renal fibrosis via blockage of mTOR in rat model and its mechanism / 解放军医学杂志

Objective To investigate the protective effects of sirolimus on unilateral ureteral obstruction (UUO) induced renal fibrosis by blockage of mTOR and its mechanism. Methods Forty-two female rats were randomized to 3 groups: UUO group, sirolimus group (Sir group), and control group. UUO rats underwent unilateral ureteral ligation to reproduce renal fibrosis model. Sir group received sirolimus 2mg/kg wt per day (0.4ml, intragastric administration) from one day before the UUO procedure to the end of study. The control group underwent surgery but without ureteral ligation. Obstructed kidneys were harvested on 7th and 14th day, and histological examination was performed for observing and comparing the degree of renal and renal tubule expansion. The concentrations of sodium, potassium and calcium ion in the urine obtained from the pelvis of the kidneys with ligated ureters were determined. At the same time, the expression of proliferating cell nuclear antigen (PCNA) and apoptosis were observed with immunohistochemical method and TUNEL respectively. MicroRNAs quantities (mir-29c, mir-143, and mir-155) were assayed by quantitative PCR. Results At abovementioned two time-points, swollen kidneys and expanded renal tubules were observed in UUO and Sir groups as compared to control group, however, kidney in Sir group showed significantly less swelling and than that in UUO group (P<0.01). Histological observation found tubular injury, cellular infiltration, and fibrosis were more marked in UUO group as compared to Sir group. Na+, K+ and Ca2+ of retention urine were significantly lower in Sir group than in UUO group (P<0.05). PCNA-positive cell ratio and apoptosis ratio were higher in UUO group than in Sir and control groups (P<0.01). No significant difference in expression of miR-155 or miR-143 was found between 3 groups, however, miR-29c expression in UUO group was down-regulated and significantly lower than that in control or Sir group (P<0.01). Conclusion With obstruction of the ureter, blocking mTOR pathways by sirolimus can attenuate fibrosis in the affected kidney.

Protective effect of sirolimus against renal fibrosis via blockage of mTOR in rat model and its mechanism / 解放军医学杂志

Objective To investigate the protective effects of sirolimus on unilateral ureteral obstruction (UUO) induced renal fibrosis by blockage of mTOR and its mechanism. Methods Forty-two female rats were randomized to 3 groups: UUO group, sirolimus group (Sir group), and control group. UUO rats underwent unilateral ureteral ligation to reproduce renal fibrosis model. Sir group received sirolimus 2mg/kg wt per day (0.4ml, intragastric administration) from one day before the UUO procedure to the end of study. The control group underwent surgery but without ureteral ligation. Obstructed kidneys were harvested on 7th and 14th day, and histological examination was performed for observing and comparing the degree of renal and renal tubule expansion. The concentrations of sodium, potassium and calcium ion in the urine obtained from the pelvis of the kidneys with ligated ureters were determined. At the same time, the expression of proliferating cell nuclear antigen (PCNA) and apoptosis were observed with immunohistochemical method and TUNEL respectively. MicroRNAs quantities (mir-29c, mir-143, and mir-155) were assayed by quantitative PCR. Results At abovementioned two time-points, swollen kidneys and expanded renal tubules were observed in UUO and Sir groups as compared to control group, however, kidney in Sir group showed significantly less swelling and than that in UUO group (P<0.01). Histological observation found tubular injury, cellular infiltration, and fibrosis were more marked in UUO group as compared to Sir group. Na+, K+ and Ca2+ of retention urine were significantly lower in Sir group than in UUO group (P<0.05). PCNA-positive cell ratio and apoptosis ratio were higher in UUO group than in Sir and control groups (P<0.01). No significant difference in expression of miR-155 or miR-143 was found between 3 groups, however, miR-29c expression in UUO group was down-regulated and significantly lower than that in control or Sir group (P<0.01). Conclusion With obstruction of the ureter, blocking mTOR pathways by sirolimus can attenuate fibrosis in the affected kidney.

Pregnancy estrogen drives the changes of T-lymphocyte subsets and cytokines and prolongs the survival of H-Y skin graft in murine model / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Estrogen as well as CD4(+)Foxp3(+) regulatory T cells were shown to have a protective role not only in maintaining maternal-fetal tolerance but also against autoimmune diseases. We aimed to investigate whether the pregnancy levels of estrogen are enough to induce transplant tolerance as to maintain fetal-maternal tolerance.</p><p><b>METHODS</b>We established H-Y skin graft transplantation in C57BL/6 ovariectomized mice that reconstituted with estrogen. Subsequently, consecutive daily estrogen injection was administrated. Tregs and the cytokines in the peripheral blood were detected by flow cytometry and ELISA pre- and post-transplant.</p><p><b>RESULTS</b>The results indicated that pregnancy levels of estrogen could promote Tregs in secondary lymphoid organs and peripheral blood (P < 0.05) but not thymus (P > 0.05). The estrogen-treated recipients accepted H-Y skin grafts for more than 35 days (median survival time (MST): (44.0 ± 1.2) days) compared with estrogen-untreated mice (MST: (23.0 ± 1.6) days) (P < 0.05). It was also observed that estrogen up-regulated the expression of Foxp3, but did not affect CD3(+)CD8(+) effector T-cells in non-transplant mice. While in the presence of H-Y antigens, the expression of Foxp3 was more significant and CD3(+)CD8(+) effector T cells were decreased significantly (P < 0.05). Meanwhile, the up-regulated IL-10 and IL-4, and down-regulated IFN-γ could be observed (P < 0.05).</p><p><b>CONCLUSIONS</b>Pregnancy levels of estrogen may promote the conversion of peripheral Tregs in secondary lymphoid organs, but show no effect on the natural Tregs production, differentiation and maturity in central lymphoid organs. Furthermore, pregnancy levels of estrogen could significantly prolong the survivals of H-Y skin grafts by the expansion of Tregs, suppression of CD3(+)CD8(+) effector T-cells and immune shift towards Th2 cytokines.</p>

Establishment and preliminary application of polymerase chain reaction with confronting two-pair primers for the singe nucleotide polymorphisms of metabolic enzymes / 中华流行病学杂志

Objective To develop a Simple,accurate,rapid,economic,large-scale detection method for the detection of singe nucleotide polymorphisms (SNPs) metabolic enzymes,using polymerase chain reaction with confronting two-pair primers (PCR-CTPP).Methods The primers of CYP1A1 (A4889G),EPHX1 (A416G) and NQO1 (C609T) were designed for PCR-CTPP,and the PCR conditions were optimized.The results of genotyping were verified by DNA sequencing.The above SNPs were detected by the PCR-CTPP detection method in a randomly selected 183 healthy individuals of Han ethnicity.The genotype frequencies were analyzed and compared with people from other ethnicities.Results The allele-specific bands of CYP1A1 (A4889G),EPHX1 (A416G) and NQO1 (C609T) were successfully amplified by PCR-CTPP under the optimal conditions and the results of genotyping were consistent with DNA sequencing.Among 183 healthy Han individuals,the genotypic distributions of CYP1A1 (A4889G),EPHX1 (A416G) and NQO1 (C609T) showed that the wild-type,homozygous variants,and heterozygotes were 103 (56.3%),8 (4.4%),72 (39.3%) and 142 (77.6%),4 (2.2%),37(20.2% ),60(32.8% ),32 (17.5%),91 (49.7%) respectively.The distributions of genotypes were all in accordance with the Hardy-Weinberg equilibrium (P＞0.05),with statistical differences and with other ethnic populations(P＜0.05).Conclusion The SNPs of metabolic enzymes can be detected by PCR-CTPP method which is simple,accurate,rapid,economic and with large scale.PCR-CTPP can be used for large scale clinical and epidemiological screening.

Pretreatment of dondor dendritic cells with Ad-IL-12p35siRNA on the survival of allograft recipients / 中华器官移植杂志

Objective To explore the effects of donor dendritic cells(DC)treated with Ad-IL- 12p35siRNA on the survival of allogragft recipients.Methods The recombinant adenoviral vectors carrying IL-12p35siRNA and HKsiRNA were transfected into bone marrow derived DC of BALB/C murine.C57BL/6 recipients were infused with DG(Ad-IL-12p35siRNA DC,Ad-HKsiRNA DC and control DC)from BALB/C donors 7 days before cardiac allograft,the survival time of murine and the change of T_H 1 and T_H2(IL-2,IL-4,IL-10 and IFN-?)cytokine were observed.Results The survival time of p35 group(20.17?2.71)days was longer than that of control group(7.81?1.61)days and HK group(7.17?1.60)days.The concentration of IL-2 and IFN-?in p35 group were significantly lower than those of control group and HK group,otherwise were the concentration of IL-4 and IL-10. Conclusion Pretreatment of dondor dendritic ceils with Ad-IL-12p35siRNA could prolonged cardiac allograft survival in recipicents.