AIM: To research the effect of serum from patients with blood stasis syndrome(BSS) associated with hypertension disease on vWF、TM、EPCR excreted by ECV-304 and Tanshinone Ⅱ_A's intervention. METHODS: Cultivated human umbilical vein endothelial cells(CRL-1730) were divided into four groups,according to different serum of patients with BSS associated with hypertension disease,BSS(model) group,non-blood stasis group,health group and control group.The incubation time was 24 h.Tanshinone Ⅱ_A was of different concentrations(40、20、10、5 ?g/mL),all Tanshinone Ⅱ_A groups' cells were incubated by patients' serum and Tanshinone Ⅱ_A for 24 h.vWF,TM and EPCR were assayed by enzyme-linked immunosorbent assay(ELISA). RESULTS: vWF,TM and EPCR secreted were higher in the model group,the non-blood stasis group and the health group than those in the control group;vWF,TM and EPCR secreted were higher in the model group,the non-blood stasis group than those in the health group;Thereinto,the difference was distinct between the model group and the health group(P

BACKGROUND: Evidence exists that inhibition of matrix metanoproteinase-2(MMP-2) secretion in the proliferating hernangioma tissue by transfection of adenovirus-active MMP-2(Ad-aMMP-2) cDNA would become an important means for treatment of proliferating hemangioma.OBJECTIVE: To investigate the influences of Ad-aMMP-2 cDNA transfection on human proliferating hemangioma growth in nude mice.DESIGN, TIME AND SETTING: A randomized, grouping, and controlled observation was performed in West China Hospital of Sichuan University between August 2003 and September 2004.MATERIALS: Eighteen BALB/c-nu/nu nude mice, weighing approximately 20 g, were included. Cavernous hemangioma specimen pathologically confirmed as proliferating hemangioma was resected from one 52-day-old female child patient.METHODS: The freshly reseoted human proliferating hemangioma specimen was sliced into small pieces with a size of 5 mm×4 mm×3 mm and subcutaneously implanted into the back of 18 nude mice within 1 hour to develop mouse models of hemangioma.Forty-five days after hemangioma implantation, 15 successful hemangioma nude mice were treated by intratumoral administration of adenovirus green fluorescent protein (Ad-GFP1 n = 51 Ad-GFP group), adenovirus-active MMP-2 (n = 5, Ad-aMMP-2 group), or the same amount of phosphate buffered saline (PBS1 n = 51 control group). Intratumoral administration was performed once every other day, for a total of 4 times.MAIN OUTCOME MEASURES: Observation of tumor volume and compadson of tumor necrosis area among 3 groups; detection of GFP expression in nude mouse; gross, hematoxylin-eosin staining, and transmission etectron microscope observation of tumor tissue morphology; determination of MMP-2 cDNA expression and microvascular density by immunohistochemistry; and detection of growth cycle and apoptosis of tumor cells by flow cytometry.RESULTS:①Ad-aMMP-2 could inhibit hemangioma growth in vivo, without marked adverse reactions. Tumor necrosis of different degrees was found in each group, and tumor necrosis area was significantly greater in the Ad-aMMP-2 group than in the control and Ad-GFP groups (P < 0.01). ②Histological sections displayed GFP gene expression in the Ad-GFP group. ③Gross observation results revealed relatively large tumor tissue in the control and Ad-GFP groups and relatively small tumor tissue in the Ad-aMMP-2 group. Hernatoxylin-eosin staining results showed that in the control and Ad-GFP groups, endothelial cells aggregated together in strip-shaped or lump-shaped appearance, and in the Ad-aMMP-2 group, there were many necrotic loci arranging in lamellar-shape appearance. Transmission electron microscope results revealed vascular endothelial cells with normal morphology in the control group and tumor cells with apparent nucleoli in the Ad-GFP group, while in the Ad-aMMP-2 group, some vascular endothelial cells exhibited chromatin pycnosis in the nucleus, forming apoptotic bodies.④ MMP-2 expression and microvascular density were significantly reduced in the Ad-aMMP-2 group than in the Ad-GFP and control groups (P < 0.05). ⑤The percentage of tumor cells in G0/G1 phase was significantly higher (P < 0.05), while the proliferating index was significantly decreased, in the Ad-aMMP-2 group than in the Ad-GFP and control groups. The Ad-aMMP-2 group exhibited higher apoptosis rate of tumor cells (P < 0.05), as well as more markedly increasing apoptosis index, than the control and Ad-GFP groups.CONCLUSION: It is feasible to block human proliferating hemangioma growth by transfeotion of Ad-aMMP-2 cDNA. The included mechanisms are to inhibit vascular endothelial cells to secrete MMP-21 thereby leading to local ischemia.

Objective To study the effects of use of cyclamic acid on monitoring D-Dimer and predicting deep venous thrombosis on lower limbs during total hip arthroplasty.Methods Ninety-three cases patients who received total hip arthroplasty operations at Joint Surgery Department of the Central Hospital of Zhuzhou city and Zhuzhou Hospital Affiliated to Xiangya Medical College of Central South University from December 2015 to May 2016 were selected as subjects and randomly assigned into study group with 50 cases and control group with 43 cases.During the operations,cyclamic acid was used intravenously and locally as a routine in the study group while saline was utilized instead in the control group.The D-Dimer was dynamically monitored before operation and 1,3,5,7,9 d after the operation,and venous color ultrasonography of both lower limbs were taken 3,6,9 d after the operation to check the conformation of thrombosis.Results The total blood loss after treatment in the study group was (350.5 ± 65.2) ml,intraoperative blood loss of (129.3 ± 43.1) ml,postoperative drainage volume of (80.9± 12.6) ml,occult blood loss of (141.9± 20.6) ml,corresponding to the control group were (560.8±60.6) ml,(208.9± 57.8) ml,(150.8 ± 18.9) ml,(202.9±23.9) ml.The above indicators were lower in the study group than in the control group,the differences between the two groups were statistically significant(t =16.02,7.59,21.24,13.22,P＜0.05).There were significant differences in terms of D2-dimer level of 1,3,5 d after the operation between the study group and the control group (P＜0.05),but at 7,9 d after the operation,the difference between the two groups were not statistically significant(P＞0.05).The study group and the control group at 9 d after operation with color Doppler ultrasound examination showed that there were 1 cases of patients with calf vein thrombosis both in two groups,there was no significant difference between the two groups(2.00％ vs.2.32％,x2 =0.012,P＞ 0.05).Conclusion The proper use of cyclamic acid can reduce blood loss and will not increase the risk of thrombosis.Monitoring dynamically on D-Dimer and deep venous color ultrasonography on lower limbs is helpful for early detection of thrombosis after total hip arthroplasty.However,the use of cyclamic acid during total hip arthroplasty will affect the monitoring on D-Dimer and therefore needs to be taken seriously.

ObjectiveTo analyze the relationship between pancreatic cancer (PC) and diabetes mellitus (DM),and the clinical and pathological features of pancreatic cancer in patients with DM.MethodsFrom January,2008 to December,2010,151 patients with PC and 195 comparable patients without PC were enrolled in a case-control study to analyze the relationship between PC and DM.ResultsThe OR was 5.91 (95％ CI 3.03-8.00,P＜0.05) in PC patients with DM for less than 2 years,and 1.308 (95％ CI 0.37-4.60,P＞0.05) in patients with DM for 2 to 5 years,and 1.16(95％ CI 0.44-3.19,P＞0.05) for patients with DM for more than 5 years.There was no significant difference between PC patients with and without DM in gender,age,body-mass index (BMD,obstructive jaundice,tumour location and tumour metastases (P＞0.05).ConclusionsThere was a significant correlation between PC and DM.De novo DM may be a clinical manifestation in patients with PC.PC patients with concurrent DM have no particular clinical and pathological features.

Chronic kidney disease (CKD) is becoming a global social problem. It is important to slow down the progression of CKD for economic and social concerns. In recent years, it has been found that colon is one of the vital organs which produce uremic toxins. And enterogenous uremic toxins are closely related to the prognosis of CKD. Theory of gut-kidney axis for the slowdown of CKD progression was raised by foreign scholars and became the research hot spot. Colon therapy with traditional Chinese medicine (TCM) has been widely used in clinical practice and is believed to slow down the progression of CKD by numerous clinical reports. However, low re-search quality and ambiguous results limited its further application. Under the guidance of senior TCM Professor Huang Chunlin, who emphasized the method of draining turbidity through bowels in the management of CKD, from the Nephrology Center, Guangdong Provincial Hospital of Chinese Medicine, as well as the modern theory of gut-kidney axis, we had carried out a series of exploratory researches which will provide data and methodology support for further confirmatory studies and improve its effectiveness.

OBJECTIVETo explore the source of small supernumerary marker chromosome in a case.

METHODSG-banded karyotyping, fluorescence in situ hybridization, multiple sequence tagged sites (STS) of the Y chromosome, and Illumima Human Cyto SNP-12 Beadchip analysis were carried out.

RESULTSThe karyotype was mos 46,X,+mar1[21]/46,X,+mar2[78]. Y chromosome STS analysis has displayed the presence of sy84, sY86, USP9Y and DDX3Y genes from the AZFa region, and sY1227 of the AZFb region, while sY1228, sY1015, sY127, sY134 from the AZFb region, and sY254 and sY255 from the AZFc region were missing. FISH analysis has verified both of the marker chromosomes to be Y chromosome fragments. Mar1 was ish.idic(Y)(q11.2)(SRY++,DXZ1+,DYZ3++,DYZ1-), while mar2 was ish.del(Y)(q11.2)(SRY+,DXZ1+,DYZ3+,DYZ1-). Single nucleotide polymorphism (SNP) microarray analysis showed that the Yq11.2-Yq12 has lost a 10.81 Mb fragment.

CONCLUSIONThe marker chromosomes were verified to be aberrant Y chromosomes, with the breakage and recombination occurring in Yq11.2. Mar 1 was an isodicentric Y chromosome (idic(Y)pter to q11.2::q11.2 to pter), and mar2 was del(Y)(q11.2). The karyotype was mos 46,X,ish idic(Y)(q11.2)(DYZ3++,SRY++,DXZ1+,DYZ1-)[21]/46,X,ish del(Y)(q11.2)(DYZ3+,SRY+,DXZ1+,DYZ1-)[78]. Combined FISH, Y chromosome STS analysis, SNP microarray analysis and other technologies can facilitate determination of the nature of marker chromosomes.

Adult ; Chromosomes, Human, Y ; genetics ; Cytogenetics ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Polymorphism, Single Nucleotide ; Sex Chromosome Aberrations ; Sex Chromosome Disorders ; genetics

OBJECTIVETo analyze the characteristics of azoospermia factor(AZF) deletions in Y-chromosome.

METHODSBased on the AZF microdeletion screening on 272 cases of azoospermia and 240 cases of severe oligozoo spermia, 49 cases were investigated using 23 sequence-tagged sites (STS) in AZFa, AZFb and AZFc. For some cases, single nucleotide rarians (SNV) method was applied to identify the single nucleotide polymorphism (SNPs) in four DAZ gene copies and to determine the copy number of the DAZ gene.

RESULTSIn 6 cases with deletions of AZFb+c, there was 1 case with sY98/sY1206 deletion, 4 cases with P5/distal-P1 recombination and 1 with P4/distal-P1 recombination. In 3 cases with deletions in AZFb, 1 case showed P5/P3 deletion and 2 cases showed P5/proximal-P1 recombination with DAZ1 and DAZ2 deletions. b2/b4 recombination was observed in all the 40 cases with deletions in AZFc. A fraction of patients with AZFb and AZFb+c deletions showed oligospermia and spermatogenic failure by testicular biopsy.

CONCLUSIONBreakpoint localization of deletions in AZF regions may help elucidating the mechanisms of microdeletions, and analysis of the characteristics and quantity of deleted genes essential for normal spermatogenesis may evaluate the association of phenotype with spermatogenic failure.

Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; Deleted in Azoospermia 1 Protein ; Gene Dosage ; Genetic Loci ; Humans ; Male ; Oligospermia ; genetics ; physiopathology ; RNA-Binding Proteins ; genetics ; Seminal Plasma Proteins ; genetics ; Spermatogenesis

OBJECTIVETo analyze chromosome aberration in a child with mental retardation and abnormalities and its parents.

METHODSChromosome G banding, multiplex ligation-dependent probe amplification, fluorescence in situ hybridization and single nucleotide polymorphisms array were employed for analysis.

RESULTSKaryotype analysis revealed that the child was 46,XX and the father was 46,XY, while the mother was 46,XX, add (12)(p13). Subtelomeric region analysis with MLPA displayed that the child has reduced ACP1 gene copy number in 2p25 region and increased SLC6A12,KDM5A gene copy numbers in 12p11 region. SNP-array has fine mapped the duplication to 12p13.33-p12.3, a 15.142 Mb region, and a deletion to 2p25.3 for 3.194 Mb, which resulted in duplication of 9 genes including SLC6A12 as well as deletion of 11 genes including SNTG2, respectively. FISH analysis revealed that the child was 46,XX,ish,der(2),t(2;12)(p25;p13)mat, or partial monosomy 2p25 and partial trisomy 12p13. In addition,the mother was a carrier with cryptic balanced translocation chromosome, 46,XX,isht(2;12) (p25;p13). Mental abnormalities and retardation of the child may be attributed to heterozygous deletion of SNTG2, MYT1L genes and duplication of SLC6A12 gene.

CONCLUSIONCombined use of MLPA, FISH and SNP-array can facilitate accurate diagnosis of cryptic rearrangement at genomic level.

Adolescent ; Adult ; Carrier Proteins ; genetics ; Child ; Child, Preschool ; Chromosome Banding ; Chromosome Deletion ; Chromosomes, Human, Pair 12 ; genetics ; Chromosomes, Human, Pair 2 ; genetics ; Female ; Gene Rearrangement ; Humans ; Intellectual Disability ; diagnosis ; genetics ; Male ; Pedigree ; Protein Tyrosine Phosphatases ; genetics ; Proto-Oncogene Proteins ; genetics ; Translocation, Genetic ; Trisomy ; Young Adult

【Objective】 To report the antibody specific identification process of a pregnant woman who had no history of blood transfusion but presented high-frequency anti-Jr^a antibodies. 【Methods】 Antibody screening and identification were performed by saline and indirect Coomb’s technique (microcolumn gel card, PEG). ABO, Rh and other blood group antigens were identified by saline. Further antibody identification tests were performed by the reaction between cells treated with various enzymes and patient plasma. Jr^a antigen was identified by human anti-Jr^a antibody. JR blood type genotyping was performed by MALDI-TOF mass spectrometry detection system. Antibody titer in serum was tested. 【Results】 The patient′s blood type was O with RhD(+ ) and CcDEe. The plasma reacted negatively with antibody screening and identification cells by saline, but positively by indirect globulin test. The self-control was negative. The patient′s Jr^a antigen was negative in serological tests and mass spectrometry blood type genotyping. Mass spectrometry revealed a homozygous nonsense mutation (c.376C>T) in exon 4. The anti-Jr^a antibody titer was 1∶2. 【Conclusion】 The patient developed high-frequency anti-Jr^a antibodies during pregnancy.

OBJECTIVE: Hypertension is a low-grade inflammation state of the disease and was easily complicated by kidneys' inflammatory response. Mangiferin (MGF), a pharmacologically active compound in various plants including Mangifera indica, has a strong anti-inflammatory activity. However, the effects of MGF on renal inflammatory injury in spontaneously hypertensive rats (SHRs) remain unclear. The purpose of this study was to investigate the protective effects and mechanisms of MGF on renal inflammatory injury in SHRs. METHODS: MGF was used in SHRs at the doses of 10, 20, 40 mg/kg/d for 8 weeks consecutively. The blood and urine were collected for assessment of renal function. Renal tissues were collected for histological, immunohistochemistry, ELISA, Western blot and real time reverse transcription PCR (RT-PCR) analysis. RESULTS: The results showed that the levels of interleukin 6 (IL-6), tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1) and recombinant chemokine C-C-Motif receptor 2 (CCR2) were increased in SHRs, meanwhile, the level of IL-10 was decreased in SHR. Treatment of MGF inhibited the expression of IL-6, TNF-α, MCP-1 and CCR2, and promoted the expression of IL-10. Furthermore, the content of blood urea nitrogen (BUN) and serum uric acid (SUA) was significantly increased in the model group, and treatment of MGF had no obvious effects on these parameters at all dose levels. CONCLUSION Our study proved that the kidneys of SHRs had significant inflammatory injury, and MGF had the protective effects on renal inflammatory injury in SHRs; The protective mechanism may be mediated partly by the MCP-1/CCR2 signaling pathway. Thus, it is a potential new drug for the treatment of hypertension.