Association of human leukocyte antigen class Ⅰ, Ⅱ allelic and haplotypic polymorphisms with leukemia in Han nationality of southern China / 白血病·淋巴瘤

Objective:To investigate the association of human leukocyte antigen (HLA) class Ⅰ (A, B and C), class Ⅱ (DRB1, DQB1 and DPB1) allelic and haplotypic polymorphisms with acute lymphoblastic leukemia (ALL), acute myeloid leukemia(AML) and chronic myeloid leukemia (CML) in Han nationality of southern China.Methods:The peripheral blood samples of 845 leukemia patients (323 cases of ALL, 350 cases of AML and 172 cases of CML) and 745 healthy blood donors from Han nationality of southern China in Shenzhen Blood Center were collected. The HLA-A, -B, -C, -DRB1, -DQB1 and -DPB1 genes were genotyped by using polymerase chain reaction-reverse sequence specific oligonucleotide (PCR-rSSO) and polymerase chain reaction-sequence based typing (PCR-SBT) methods to identify the first 4 digits of HLA alleles. The Arlequin 3.5 software was used to analyze HLA haplotypes. The correlations between HLA allelic and haplotypic polymorphisms and three types of leukemia were statistically analyzed at HLA low-resolution level (the first 2 digits of alleles) and high-resolution level (the first 4 digits of alleles), respectively.Results:P-values were adjusted by Bonferroni correction. In ALL group, the frequencies of A*02 (36.22% vs. 28.26%, χ 2 = 13.41, PC < 0.01) and its haplotype A*02-B*46-C*01 (15.35% vs. 10.23%, χ 2 = 10.90, PC = 0.02), DRB1*12 (15.79% vs. 11.10%, χ 2 = 9.02, PC = 0.03), A*02:03 (9.75% vs. 5.32%, χ 2 = 14.25, PC = 0.002) and its haplotype A*02:03-B*38:02-C*07:02 (3.80% vs. 1.51%, χ 2 = 10.41, PC = 0.02) were higher than those in healthy controls, which implied that these factors could confer risk effect in ALL. In AML group, the frequency of A*11-B*15-C*08-DRB1*15-DQB1*06-DPB1*02 (1.34% vs. 0.07%, χ 2 = 12.54, PC = 0.003) was significantly higher than that in healthy controls, suggesting that the risk effect might be conferred by this haplotype. In CML group, the frequencies of A*02 (36.63% vs. 28.26%, χ 2 = 9.33, PC = 0.02) and its haplotype A*02-B*15-C*04 (2.17% vs. 0.29%, χ 2 = 11.74, PC = 0.02), and DRB1*03:01-DQB1*02:01-DPB1*02:01 (1.86% vs. 0.14%, χ 2 = 13.10, PC = 0.01) were higher than those in healthy controls, which implied that these factors could confer risk effect in CML, whereas the frequency of DRB1*13 (1.45% vs. 5.25%, χ 2 = 9.29, PC = 0.03) was lower than that in healthy controls, suggesting that it was a CML antagonistic gene. Conclusion:Leukemia susceptible or antagonistic HLA alleles and haplotypes are found at low-resolution and high-resolution levels of HLA, which might provide reference for investigating the pathogenesis of leukemia and guiding formulation of effective treatment strategies in Han nationality of southern China.

Identification of a novel KIR3DL3*064 allele by cDNA cloning and sequencing / 中华医学遗传学杂志

OBJECTIVE@#To report on a novel KIR3DL3 allele identified in a southern Han Chinese individual.@*METHODS@#Peripheral blood sample was collected from a voluntary blood donor with inconclusive result by KIR3DL3 sequence-based typing (SBT). Total mRNA was extracted and subjected to reverse transcription to obtain KIR3DL3 cDNA, which was then amplified by PCR with a pair of KIR3DL3-specific primers. The product was subjected to cDNA cloning and sequencing.@*RESULTS@#cDNA cloning and sequencing have identified a wide-type KIR3DL3*00802 allele and a novel KIR3DL3*064 allele. The latter differed from KIR3DL3*00601 by a missense variant at codon 374[c.1184 C>T (p.Thr374Ile)] in exon 9. The novel KIR3DL3 allele has been officially assigned by the KIR subcommittee of World Health Organization Nomenclature Committee for factors of HLA system.@*CONCLUSION@#cDNA cloning and sequencing may be used to distinguish inconclusive results in KIR3DL3 SBT in order to identify novel KIR alleles.

Allelic diversity of KIR2DL4 gene and identification of five novel alleles among southern Han Chinese population / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study the polymorphisms of the KIR2DL4 gene in a southern Han Chinese population.</p><p><b>METHODS</b>Genomic DNA isolated from 306 unrelated individuals was amplified by using KIR2DL4-specific PCR primers. The PCR products were genotyped for the entire coding sequence by sequencing-based typing (SBT). Assignment of allelic profile was accomplished with an Assign 3.5 software. For samples with inconclusive SBT results, the RT-PCR products covering the entire coding sequence of the KIR2DL4 gene were subjected to cloning and haplotype sequencing.</p><p><b>RESULTS</b>Among the 306 individuals, 11 alleles were detected, of which 5 novel alleles were officially named by the KIR subcommittee of the World Health Organization Nomenclature Committee for factors of HLA system. The observed frequencies for the 11 alleles were KIR2DL4 *00102 (75.5%), *00103 (8.2%), *00501 (34.0%), *00503 (0.7%), *00504 (0.7%), *00602 (14.4%), *00801 (11.4%), *011 (22.2%), *032 (0.3%), *033 (0.3%) and *034 (0.3%). The ratio of 10A type alleles including 2DL4*00102, *00103, *00501, *00503, *00504, *00602, *032, *033, *034 and 9A type alleles including 2DL4*00801, *011 were 97.0% and 33.0%, respectively, with a ratio of 2.9:1.</p><p><b>CONCLUSION</b>The allelic diversity of the KIR2DL4 gene in southern Han Chinese has been elucidated, which may provide valuable data for research on transplantation, reproductive immunity, KIR-associated disease and evolution.</p>

Genetic polymorphisms of KIR2DS4 gene among ethnic Hans from southern China / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To study genetic polymorphisms of the KIR2DS4 gene among ethnic Hans from southern China.</p><p><b>METHODS</b>Genomic DNA was isolated from 306 unrelated individuals and amplified with KIR2DS4-specific PCR primers. KIR2DS4-positive samples were genotyped for the entire coding sequence by sequencing-based typing (SBT). Assignment of allelic genotypes was accomplished by using Assign 3.5 software. For samples with inconclusive SBT results, RT-PCR products covering the entire coding sequence of the KIR2DS4 gene were subjected to cloning and haplotype sequencing.</p><p><b>RESULTS</b>Among all tested samples, 297 were demonstrated to have carried the KIR2DS4 framework gene. For KIR2DS4-positive samples subjected to SBT for the entire coding sequences, no background was observed with the obtained sequences. Three of the seven identified alleles were of novel types, which were officially named by the KIR subcommittee of the World Health Organization Nomenclature Committee for Factors of HLA System. The observed frequencies for the 7 alleles were KIR2DS4*00101 (78.8%), *003 (10.5%), *004 (16.0%), *010 (23.2%), *017 (0.3%), *00105 (0.3%) and *018 (0.7%), respectively. Allele KIR2DS4*007 was not found. The overall frequency for normal cell-surface expression KIR2DS4 alleles including 2DS4*00101, *017 and *00105 was 79.4%, and that for non cell-surface expression alleles including 2DS4*003, *004, *010 and *018 was 50.4%. The ratio between the two was 1.6:1.</p><p><b>CONCLUSION</b>The present study has elucidated the allelic diversity of KIR2DS4 among ethnic Hans from southern China, which may provide valuable data for transplantation as well as studies on KIR-associated disease and evolution.</p>

Preliminary study on polyetheretherketone implant applied to cranioplasty of skull defect / 天津医药

Objective To study on the procedure, safety and effectiveness of polyetheretherketone (PEEK) implant applied to cranioplasty of skull defect. Methods A total of 11 cases (10 male, 1 female) of unilateral skull defect, more than 6 months post operation, were included in this study. PEEK implant was custom-made by three-dimensional numerically controlled processing depended on the data obtained from 1 mm-slice CT scan before cranioplasty individually. Autoclaved implants were applied to cranioplastic surgeries under general anesthesia. Findings of imaging examination and vital signs were compared before and after operation. Vital sign changes and circumstances during procedure were noted, and following-up reviews were performed on 2-week, 3-month and 1 year after operation respectively. Results Wounds healing were uneventful in 11 cases, no postoperative paresthesia presented. Surgical complications including temporary subcutaneous exudates were cleared up by puncture and aspiration in 5 cases, subcutaneous hematoma duo to drainage removal in 1 case, of which an emergency evacuation was performed and the implant was still with instant incision closure and then smooth recovery eventually. No significant abnormal fluctuation of laboratory examination was reported, moreover no artifact interference was found on postoperative CT scan or MR image. The subjective feelings and external sensory effects are satisfactory in patients. No emerging dysfunction of central nervous system or other organs was found, and no long-term complication was appeared. Conclusion It is feasible and safe to apply PEEK implant to cranioplasty without additional operative difficulty. This kind of material is an ideal alternative for repairing skull defect to patients with good financial condition and specific demands for postoperative status especially.

Cloning and sequencing of KIR2DL1 framework gene cDNA and identification of a novel allele / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To develop an assay for cDNA cloning and haplotype sequencing of KIR2DL1 framework gene and determine the genotype of an ethnic Han from southern China.</p><p><b>METHODS</b>Total RNA was isolated from peripheral blood sample, and complementary DNA (cDNA) transcript was synthesized by RT-PCR. The entire coding sequence of the KIR2DL1 framework gene was amplified with a pair of KIR2DL1-specific PCR primers. The PCR products with a length of approximately 1.2 kb were then subjected to cloning and haplotype sequencing.</p><p><b>RESULTS</b>A specific target fragment of the KIR2DL1 framework gene was obtained. Following allele separation, a wild-type KIR2DL1*00302 allele and a novel variant allele, KIR2DL1*031, were identified. Sequence alignment with KIR2DL1 alleles from the IPD-KIR Database showed that the novel allele KIR2DL1*031 has differed from the closest allele KIR2DL1*00302 by a non-synonymous mutation at CDS nt 188A>G (codon 42 GAG>GGG) in exon 4, which has caused an amino acid change Glu42Gly. The sequence of the novel allele KIR2DL1*031 was submitted to GenBank under the accession number KP025960 and to the IPD-KIR Database under the submission number IWS40001982. A name KIR2DL1*031 has been officially assigned by the World Health Organization (WHO) Nomenclature Committee.</p><p><b>CONCLUSION</b>An assay for cDNA cloning and haplotype sequencing of KIR2DL1 has been established, which has a broad applications in KIR studies at allelic level.</p>

A strategy to clarify ambiguities during genotyping of functional KIR framework genes by sequencing-based typing among ethnic Hans from southern China / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To categorize ambiguous allelic combinations encountered in genotyping of functional KIR genes by sequencing-based typing of the entire coding sequence and develop an efficient approach to identify such ambiguities.</p><p><b>METHODS</b>Fourteen KIR functional framework genes from 306 ethnic Chinese individuals were genotyped for the entire encoding sequence. The number of ambiguities was directly counted. Based on the differences within each ambiguous allelic combination, group-specific PCR primers and sequencing primers for amplifying the target allelic sequence were designed. The PCR products were then subjected to sequencing in order to identify the ambiguities.</p><p><b>RESULTS</b>The 14 functional KIR genes were subjected to sequencing-based typing (SBT) for the entire encoding sequence. Six ambiguous allelic combinations were identified. The most common ambiguity 3DL2*(002, 007/010, 015) has accounted for 12.09% of the 306 tested samples. The remaining 5 ambiguities were (2DL5A*001, 2DL5B*006/2DL5A*012, 2DL5B*008), 3DL3*(001, 010/009, 048), 3DL2*(007, 008/016, 027), 3DL3*(00801, 048/01001, 026) and 3DL3*(00802, 048/01002, 026) have accounted for 5.88% (18/306), 3.59% (11/306), 2.29% (7/306), 1.31% (4/306) and 1.31% (4/306) of all samples, respectively. For two ambiguities (2DL5A*001, 2DL5B*006/2DL5A*012, 2DL5B*008) and 3DL2*(007, 008/016, 027) subjected to group-specific PCR and re-sequencing, only one demonstrable genotype was identified, While for in each of other four ambiguities subjected to group-specific PCR and re-sequencing, two different genotypes were identified.</p><p><b>CONCLUSION</b>An efficient approach by group-specific PCR and sequencing retest has been established to clarify the ambiguities during SBT testing for functional KIR framework genes, which may have a broad application in KIR sequencing-based typing.</p>

Effects of simvastatin versus alendronate on bone loss and biomechanical property deterioration in ovariec-tomized rats / 医学研究生学报

Objective Alendronate is widely used as an anti-osteoporosis agent , while simvastatin , as a lipid-lowering drug , is being considered beneficial for bone formation .The present study aims to compare the effects of simvastatin and alendronate on bone loss in ovariectomized ( OVX) rats. Methods Thirty-two female SD rats were equally randomized into a sham operation , an OVX mod-el, a simvastatin ( OVX +S), and an alendronate ( OVX +A) group.All the rats but those in the sham operation group underwent dual ovariectomy .The animals of the OVX model and OVX +S groups were treated intragastrically with vehicle and simvastatin at 5 mg per kg of the body weight per day , respectively , while those of the OVX+A group injected subcutaneously with alendronate at 70μg per kg of the body weight per week .After 12 weeks of intervention , all the rats were sacrificed for detection of the serum concentrations of PINP and ICTP by ELISA, analysis of bone mineral density and bone histo-morphometry parameters of L 4 vertebrae , determination of the maximum loading and elastic modulus of L 5 vertebrae by compression test. Results The serum concentrations of P1NP and ICTP were significantly lower in the sham operation than in the other three groups (P0.05).Bone histomorphometry showed significantly lower BV /TV in the OVX model than in the sham operation and OVX +A groups ([19.9 ±1.69]%vs [29.03 ±2.59]%and [27.05 ±1.91]%, P<0.05), but markedly higher Tb.Sp in the former than in the latter two groups ([357.33 ±26.55] μm vs [211.17 ±16.56] μm and [252.50 ±19.35] μm, P<0.05).The maximum load and elastic modulus of L5 vertebrae were significantly lower in the OVX model rats than in the other three groups (P <0.05). Conclusion Both simvastatin and alendronate can inhibit bone loss in OVX rats , with comparable effects of preventing the deteriora-tion of biomechanical properties , but the latter is evidently more effective in maintaining bone mineral density .

Effect of simvastatin on bone mineral density and biomechanical properties of ovariectomized rats / 中国组织工程研究

BACKGROUND:Osteoporosis and its complications severely threaten the elder’s health. Simvastatin, widely accepted as a lipid-lowering drug, is reported to potentialy promote bone formation, but it is in debate when oraly administered, and there is no evidence to support whether this is due to the region difference. OBJECTIVE:To investigate the effect of oraly administered simvastatin on bone mass and biomechanical properties of the femur and vertebrae in osteopenia rats induced by ovariectomy (OVX). METHODS: Twenty-four 6-month-old female Sprague-Dawley rats were subjected to OVX+oraly administered saline vehicle (OVX group,n=8), OVX+oraly administered simvastatin (5 mg/kg/d; intervention group,n=8) or sham surgery (sham group,n=8). After 8 weeks of treatment, al rats were sacrificed and the level of procolagen type I N-terminal propeptide in blood serum was assessed by ELISA. Bone mineral density was determined in the L5 vertebra and left femur using dual-energy X-rays. Furthermore, the biomechanical properties of the L4 vertebra and right femur, including maximum load and elastic modulus, were detected by compression testing and three-point bending test, respectively. RESULTS AND CONCLUSION: The serum level of procolagen type I N-terminal propeptide in the sham group was significantly lower than that in the other two groups. OVX rats showed significantly lower bone mineral density in both the L5 vertebra and left femur than sham rats (P < 0.05). Rats in the intervention group showed higher bone mineral density than those in the OVX group, with statisticaly significant difference in the L5 vertebra (P < 0.05), but insignificant difference in the femur. Maximum load and elastic modulus of the L4 vertebra in the OVX group were significantly lower than those in the sham and intervention group. Markedly lower elastic modulus of the femur was found in the OVX group than the sham and intervention groups. These findings demonstrate that simvastatin treatment can partialy prevent bone loss in OVX rats with more notable effect on the vertebrae than the femur, and for this model, the vertebra is superior to the femur used in biomechanical test.

Simvastatin effects on the expressions of specific osteogenic genes in bone marrow stromal stem cells / 中国组织工程研究

BACKGROUND:Previous studies have demonstrated that simvastatin that can promote osteogenic differentiation of bone marrow stromal stem cel s in vitro, is likely to be a new osteogenic drug. While it is stil unknown whether there is time-dependent stimulation of simvastatin on the expressions of bone morphogenetic protein 2 and col agen type I. OBJECTIVE:To investigate the expressions of bone morphogenetic protein 2 and col agen type I in rat bone marrow stromal stem cel s in vitro stimulated by simvastatin at different time points. METHODS:Passage 1 bone marrow stromal cel s were divided into control and simvastatin group, fol owed by cultured in osteogenic differetiation medium with or uithout 10-7 mol/L simvastatin. After 7-day intervention, expression of alkaline phosphatase was detected in passage 3 cel s. Passage 4 cel s were divided and cultured as described above, and afterwards, RNA and proteins were extracted at 12 and 36 hours to detect the expressions of bone morphogenetic protein 2 and col agen type I using real-time PCR and western blot assay. RESULTS AND CONCLUSION:Both two groups could express alkaline phosphatase, while the rate of positive cel s significantly increased in the simvastatin group compared with the control group (P<0.05);at 12 and 36 hours after intervention, mRNA expressions of bone morphogenetic protein 2 and col agen type I in the simvastatin group were significantly higher than those in the control group (P<0.05). Besides, western blot assay showed:at both 12 and 36 hours, simvastatin significantly enhanced the expression of bone morphometric protein 2, while the expression of col agen type I significantly increased at 12 hours (P<0.05), but not at 36 hours. In conclusion, simvastatin can promote the expressions of bone morphometric protein 2 and col agen type I in rat bone marrow stromal cel s, with more favorable outcomes after 12-hour treatment.

Role of ERS in Astragaloside Ⅳ-induced cardioprotection against ischemia/reperfusion injury in rats / 中国药理学通报

Aim To explore the role of endoplasmic re-ticulum stress( ERS) in Astragaloside Ⅳ-induced car-dioprotection against ischemia/reperfusion injury in rats. Methods A model of myocardial ischemia 30 min followed by 120 min reperfusion was made by liga-ting coronary artery in male Wistar rats. Rats were di-vided randomly into 4 groups: sham group, ischemia/reperfusion group, ERS inhibitor TUDCA group, As-tragaloside Ⅳgroup. Myocardial samples were collect-ed from the risk zones during ischemia and reperfu-sion, ERS was determined by measuring levels of glu-cose regulated protein 78 ( GRP78 ) , an established marker of ERS with Western blot. Immunofluorescence study was used to test GRP78 intensity with laser scan-ning confocal microscopy, TTC method was used to measure the infarct size,hematoxylin-eosin staining was used to observe the changes of morphological changes of myocardium. Results There was no statistical difference in GRP78 expression during ischemia com-pared to the sham group, but was markedly increased upon reperfusion. Astragaloside Ⅳ could mimic TUD-CA and significantly decreased the GRP78 expression, reduced infarct size and improved the morphology of myocardial tissue with a significant statistical difference compared with the control group ( P<0. 05 ) . Conclu-sions ERS is induced upon reperfusion but not during ischemia in isolated rat hearts. Astragaloside Ⅳ pre-vents myocardial reperfusion injury presumably by the inhibition of ERS.

Effects of simvastatin treatment with different intervention programs on the bone quality of osteoporotic rats / 中国比较医学杂志

Objective To investigate the effects of simvastatin on the bone loss and deterioration of bone quality with different intervention programs. Methods Thirty two 3?month?old female Sprague?Dawley ( SD ) rats were randomized into 4 groups of 8 animals in each: All rats but those in the sham group ( A) received bilateral ovariectomy, and treated by vehicle (B), simvastatin (5 mg/kg/day) at first half period (C) or at latter half period (D). The rats in group C received simvastatin by a gavage after the OVX operation immediately, and stopped at 10 weeks after OVX. The rats in group D began to receive simvastatin treatment from 10 weeks after OVX and ended at 20 weeks after OVX. At week 20, all rats were sacrificed and the concentrations of serum PINP and ICTP were detected by ELISA, L3 vertabra was used to detect the bone mineral density, L4 vertebra was used to analyze the maximum loading and elastic modulus by compression test, and the microstructure of the L5 vertebra was detected by micro?computed tomography. Results 1. ELISA analysis:the concentrations of serum P1NP and ICTP in the groups A, B and C were significantly higher than that of group A (P<0?05). 2. BMD test showed that the rats in group B had significantly lower BMD than the other 3 groups (P<0?05), while the BMD of groups C and D were markedly lower than that of group A (P<0?05). 3. Biomechanical test:the maximum load and elastic modulus of L4 vertebrae in the group B were significantly lower than the other 3 groups ( P<0?05), and those of the groups C and D were markedly lower than that in the group A (P<0?05). 4. micro?CT:BV/TV and Tb. N in the sham operated rats were significantly higher than those of the other 3 groups, while the opposite trends was found in Tb. Sp (P<0?05), and the Tb. Sp in the groups C and D were significantly lower than that of group B (P<0?05). Conclusions Our data demonstrate that bone loss and deterioration of bone micro?structure and biomechanical properties occurred at 20 weeks after ovariectomy, both the first?half?period and latter?half?period treatment by simvastatin may partially prevent these changes, but can not restore the BMD to normal level.

Effect of bortezomib on migration and invasion in cervical carcinoma HeLa cell

Objective: To explore the effect of bortezomib on migration and invasion of cervical carcinoma HeLa cell and specific molecular mechanism. Methods:The effect of bortezomib on the viability of HeLa cell was measured by MTT assay. The effect of bortezomib on cell migration and invasion was measured by Transwell assay and invasion experiment respectively. The activation of Akt/mTOR signaling pathway and expression level of MMP2, MMP9 were assayed by western blot. Results:MTT assay indicated bortezomib (2.5μM, 5μM, 10μM) could inhibit HeLa cell viability, and the inhibitory rate was highest at 48 h. Transwell assay and invasion experiment results showed that bortezomib inhibited HeLa cell migration and invasion. Western blotting assays presented bortezomib could suppress the phosphorylation of Akt and mTOR, and down-regulate the expression of MMP2 and MMP9. Conclusions:These results suggested bortezomib could inhibit migration and invasion in cervical carcinoma HeLa cell, which might be related to Akt/mTOR signal pathway.

Expression of matrix metalloproteinase-3 and tissue inhibitor of metalloproteinase 1 in knee articular cartilage / 中国组织工程研究

BACKGROUND:Tissue inhibitor of matrix metaloproteinase 1 (TIMP-1) is the corresponding antagonist of matrix metaloproteinase 13 (MMP-13), and their balance between expression and functional activity exerts an important role in the metabolic state of the extracelular matrix. During the development of osteoarthritis, however, TIMP-1 and MMP-13 expressions and their expression ratio show unclear changes in DH guinea pigs. OBJECTIVE:To explore the expression levels of MMP-13 and TIMP-1 in DH guinea pigs with different ages, and to analyze the relationship between the ratio of MMP-13 to TIMP-1 and the age-dependent degenerative changes in the articular cartilage. METHODS:Twenty-four female Dunkin Hartley guinea pigs were sacrificed at age of 2, 4, 8, 12 months separately, with six animals at each time point. The knee joints were colected and gross visual appearance of the articular cartilage was observed, then were decalcified and prepared for paraffin sections. VG staining and Mankin score were used to analyze the histological changes. Immunohistochemistry was conducted to assess the expression levels of MMP-13 and TIMP-1 in the cartilage. Integrated absorbance values were used as the quantitive analysis calculated by Image pro-Plus 6.0. Linear regression analysis was done to analysis the relationship between Mankin score and the ratio of MMP-13/TIMP-1. RESULTS AND CONCLUSION:Normal appearance in the articular cartilage was observed in 2-month-old DH guinea pigs, while degenerative changes in the articular cartilage were shown in 4-month-old animals, which became severer with age. Significant difference was found in Mankin score between any two groups (P < 0.05). The ratio of MMP-13 to TIMP-1 increased with age, and the ratio was positively correlated to the Mankin score (P < 0.05). Age-related articular cartilage degeneration occurred in Dunkin Hartley guinea pigs at 4 months of age, and devoloped with age, which is related with the imbanlance of the expression ratio of MMP-13 to TIMP-1.

Human telomerase reverse transcriptase gene-modified bone marrow mesenchymal stem cell transplantation for cerebral infarction / 中国组织工程研究

BACKGROUND:Studies have shown that human telomerase reverse transcriptase (hTERT) has the ability to enhance cel proliferation, maintain telomere length, prolonged cel life cultured in vitro. OBJECTIVE:To observe the effect of hTERT gene-modified bone marrow mesechymal stem cel transplantation on neural function recovery of rats with cerebral infarction. METHODS:Rat models of middle cerebral artery occlusion were established and randomized into model group, cel transplantation group and hTERT-modified cel transplantation group, with 20 rats in each group. Rats in the three groups were respectively injected via tail vein with 1 mL PBS, passage 9 bone marrow mesenchymal stem cel suspension (2.5×107/L) and hTERT-modified passage 9 bone marrow mesenchymal stem cel suspension (2.5×107/L), respectively. Modified neurological severity scores were determined before and after transplantation; RT-PCR and western blot assay were used to measure hTERT expression at gene and protein levels; TUNEL method was adopted to detect cel apoptosis in the brain. RESULTS AND CONCLUSION:hTERT-modified bone marrow mesenchymal stem cels had prolonged cel cycle, and with the increase in passage number, the cels showed good growth with no changes in morphology. The expressions of hTERT mRNA and protein were superior in the hTERT-modified cel transplantation group than the cel transplantation group, and there was a significant difference (P < 0.05). Modified neurological severity scores and number of apoptotic cels were decreased significantly in the hTERT-modified cel transplantation group compared with the other two groups (P < 0.05). These findings indicate that hTERT-modified bone marrow mesenchymal stem cels can promote neural functional recovery of rats with cerebral infarction.

Hyperbaric oxygen therapy improves nerve regeneration microenvironment and promotes rat nerve function recovery after cerebral infarction / 中国组织工程研究

BACKGROUND:Numerous clinical studies have confirmed that the microenvironment at a spinal cord injury site can be obviously improved through hyperbaric oxygen therapy; however, what effect does hyperbaric oxygen have on the microenvironment of the injured brain? OBJECTIVE:To investigate the effect of hyperbaric oxygen therapy on nerve regeneration microenvironment and the recovery of rat nerve function after focal cerebral infarction. METHODS:Rat models of focal cerebral infarction were established by occlusion of the middle cerebral artery and subjected to hyperbaric oxygen therapy. Sham group and model group were established as comparison. In the sham group, rat models of focal cerebral infarction were established but did not receive any treatment. Rats in the model group were placed in a hyperbaric oxygen therapy chamber but the pressure and oxygen concentration were not administered. RESULTS AND CONCLUSION:Compared with the model group, the score of rat limb function at 16 days after treatment and the expression of growth associated protein 43 in the rat cerebral infarcted area at postoperative 14 days were significantly increased , but infarct volume at postoperative 24 hours was al significantly decreased in the hyperbaric oxygen therapy group (alP < 0.05). These results confirmed that hyperbaric oxygen therapy can improve nerve regeneration microenvironment and promote the recovery of rat nerve function after focal cerebral infarction.

Effects of α1-PDX, a furin inhibitor, on growth, invasion, and tumorigenicity of cervical cancer HeLa cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of the furin inhibitor α1-PDX on the growth, invasion, and tumorigenicity of cervical cancer cells and explore the mechanisms.</p><p><b>METHODS</b>The changes in the growth, migration and invasion of α1-PDX-transfected HeLa cells were observed using MTT assay, Boyden migration and invasion assay. The protein levels of furin and MT1-MMP were measured using Western blotting and furin activity was detected by enzyme activity assay in the transfected cells. HeLa cells were seeded subcutaneously in nude mice and the tumor volume changes were recorded.</p><p><b>RESULTS</b>Compared with the control cells, α1-PDX-treated cells showed a significant growth inhibition by 18.4% at 24 h (P<0.01) with obviously lowered migration ability and cell invasiveness (P<0.01). Treatment with α1-PDX significantly reduced furin enzyme activity and MTI-MMP protein levels in HeLa cells. In nude mice, α1-PDX-treated HeLa cells exhibited a delayed and lowered tumorigenicity with reduced size of the tumors.</p><p><b>CONCLUSION</b>α1-PDX can inhibit the growth, metastasis and tumorigenicity of HeLa cells, the mechanism of which may involve a decreased furin activity and MTI-MMP expression.</p>

The Application of Fourier Transform Infrared Spectrometer to theSmoke Determination / 分析化学

The smoke of anti-infrared smoke agent in a self-made miniature smoke chamber was determined by using Forier transform infrared spectrometer. The smoke changing and settling regulation with time were obtained by measuring the IR spectra of smoke at different time dynamically. Through testing the smoke of different quantity of the same smoke agent, it was found that the relationship between the concentration of the main smoke products and the absorbance obeyed Lambert-Beer′s Law. Finally, the relationship between the smoke screening ability and reactant agent amounts was obtained by calculating the screening ability of different amounts of reactant agent in three “atmosphere windows”.

Effects of hypoxia on expression of VEGF, iNOS and eNOS mRNA in human umbilical vein endothelial cells from Tibetan and Han / 中国病理生理杂志

AIM: To compare the effects of hypoxia on expression of vascular endothelial growth factor (VEGF), iNOS and eNOS mRNA in cultured umbilical vein endothelial cells (UVECs) obtained from Tibetan and Han. METHODS: UVECs were obtained from native Tibetan and immigrant Han, respectively and cultured under hypoxia conditions (0.5% oxygen) for 2 h, 4 h, 12 h, and 24 h and normoxic conditions. VEGF, iNOS and eNOS mRNAs were detected with methods of RT-PCT. RESULTS: VEGF and iNOS mRNAs were up-regulated while eNOS mRNA depressed by hypoxia similarly in Tibetan and Han UVECs. CONCLUSION: Our results suggest that the changes of VEGF, iNOS and eNOS mRNA expression are common pathways in the mechanisms of hypoxic responses.