OBJECTIVETo identify the fibrinogen (Fg) gene mutations in a Chinese pedigree of congenital afibrinogenemia.

METHODSThe plasma Fg activity and protein of the proband and his family members were detected. Genomic DNA was isolated from the peripheral blood mononuclear cells. All the exons and exon-intron boundaries of fibrinogen gene were amplified by PCR and sequenced thereafter.

RESULTSTwo mutations, 7972 del G in FGB and T2543A in FGG, were found in the proband.

CONCLUSIONSFGG2543 is a polymorphism site, which lead to the polymorphism of gamma144 I/K. The G deletion at base 7972 of FGB contributes to the frameshift mutation after amino acid 419, resulting in the truncated beta chain without the terminal 27 amino acids. The latter may contributes to the pathogenetic mechanisms in Chinese congenital afibrinogenemia patients. The G deletion at base 7972 of FGB is identified for the first time.

Adult ; Afibrinogenemia ; congenital ; genetics ; metabolism ; Base Sequence ; Blotting, Western ; DNA Mutational Analysis ; Exons ; genetics ; Female ; Fibrinogen ; genetics ; Humans ; Introns ; genetics ; Male ; Mutation ; Pedigree ; Polymerase Chain Reaction

AIMTo design and synthesize new methoxyl carbazole analogues as antitumor compounds.

METHODSMethoxyl-nitrobiphenyls (3a-3c) were prepared through the Ullmann reaction of 4,5-dimethoxyl-2-bromonitrobenzene and methoxyl-iodobenzene compounds with the catalysis of copper powder, and then reduced by P(EtO)3 to obtain methoxyl carbazoles 4a-4c. The modification at 9-position of the methoxyl carbazoles (4a-4c) gives 16 carbazole derivatives (5a-5p). These compounds were confirmed by 1HNMR, MS, IR and elemental analysis.

RESULTIn vitro antitumor activities evaluation in vitro demonstrated that IC50 value of the target compounds 4c, 5a, 5b, 5g, 5h, 5i, 5l, 5n and 5p against HT-29 cells were 12.1, 10.6, 8.1, 3.1, 4.4, 10.1 and 9.2 micromol x L(-1) respectively, and IC50 value of the target compound 4a against KB was 17.7 micromol x L(-1).

CONCLUSIONSome of the target compounds have better inhibitory effects against H-29 and KB cells.

Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Carbazoles ; chemical synthesis ; chemistry ; pharmacology ; HT29 Cells ; drug effects ; Humans ; KB Cells ; drug effects ; Molecular Structure

OBJECTIVETo compare the effect of autograft, cage and autograft plus internal fixation with plate on maintenance of disc height and recovery of spinal cord function.

METHODSFrom March 1998 to June 2004, 117 cervical spondylotic patients who received anterior fusion surgery were reviewed. There were 65 males and 52 females with an average age of 57 years (range from 31 to 72 years). One level was involved in 91 patients, and two levels in 26. Fusion methods included autogenous tri-cortical iliac bone grafting alone in 31 patients, cage with autograft bone in 38 and autogenous tri-cortical iliac bone grafting combined with self-locking plate in 49. The measured fusion length was determined as the distance from the midpoint between the anterior and posterior aspects of the superior endplate of the top level of the fusion to the midpoint of the inferior endplate of the bottom level of the bottom level of the fusion. JOA rating criteria was used for cord function evaluation. One-way variation analysis was used to compare the disc height loss and cord function among the 3 groups.

RESULTSAll patients were followed-up for an average period of 23 months (range, 13 to 59 months). At final follow- up, disc height loss averaged 1.94 mm in autogenous tri-cortical iliac bone grafting group, greater than that in the other two groups (1.48 mm in cage group and 1.25 mm in instrumented group) with statistical significance. Recovery of spinal cord function was satisfactory in all groups at one year postoperatively with no statistical difference. Three implants failure occurred in self-locking plate group and was salvaged with uneventful recovery.

CONCLUSIONSIn anterior cervical fusion surgery, maintenance of disc height could be achieved by proper application of cage or internal fixation with plate.

Adult ; Aged ; Bone Plates ; Cervical Vertebrae ; pathology ; surgery ; Decompression, Surgical ; instrumentation ; methods ; Female ; Humans ; Ilium ; transplantation ; Male ; Middle Aged ; Retrospective Studies ; Spinal Fusion ; instrumentation ; methods ; Spinal Osteophytosis ; surgery ; Transplantation, Autologous ; Treatment Outcome

Adult ; Arthroscopy ; methods ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged ; Shoulder Joint ; diagnostic imaging ; pathology ; surgery ; Tomography, X-Ray Computed ; Treatment Outcome

This study was purposed to analyze and identify a novel HLA allele in Chinese population. A new HLA-B allele which is closely related to HLA-B*35:03:01 was initially detected by PCR-SSOP, then DNA sequencing was performed to identify the difference between the novel allele and HLA-B*35:03:01 allele. The result showed that the sequence of the new allele was different from all other known sequence. It differs from the closest matching HLA-B*35:03:01 by a single substitution at position 387 C→G in exon 3, no resulting in amino acid change. It is concluded that this allele is a novel one and has been officially named B*35:03:07 by the WHO Nomenclature Committee.
Alleles ; Asian Continental Ancestry Group ; genetics ; HLA-B Antigens ; genetics ; Humans ; Male ; Sequence Analysis, DNA

Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Hepatitis B ; epidemiology ; Humans ; Infant ; Middle Aged ; Seroepidemiologic Studies ; Young Adult

BACKGROUND: The preliminary study found that domestic porous tantalum is conducive to the early adhesion and proliferation of MG63 cells, which can be used as a scaffold material for bone tissue engineering. As an optimized product of platelet-rich plasma, platelet lysate is more suitable for bone induction in the bone repair. OBJECTIVE: To further investigate the effect of platelet lysate and domestic porous tantalum scaffold constructs on the proliferation of MG63 cells and expression of integrin β1 (ITGβ1)/Vinculin/F-actin signaling pathway based on our previous findings. METHODS: MG3 cells were cultured and inoculated onto domestic porous tantalum scaffolds with the addition of 3%, 5%, 7% and 10% platelet lysates. Then, 7% as the best volume fraction of platelet lysate was screened by cell counting kit-8 method. There were four experimental groups including blank group (normally cultured MG63 cells), platelet lysate group (MG63 cells were cultured in 7% platelet lysate), porous tantalum scaffold group (MG63 cells were cultured on the domestic porous tantalum scaffold), and combined group (MG63 cells were cultured with 7% platelet lysate and porous tantalum scaffold. Scanning electron microscope was used to observe the surface morphology of domestic porous tantalum and platelet lysate-porous tantalum scaffold-MG63 cell complex. Cell counting kit-8 method was used to detect the proliferation of MG63 cells. Real-time fluorescence quantitative PCR (qPCR), immunocytochemical staining and western blot were used to detect the expression of ITGβ1, Vinculin, F-actin in MG63 cells at mRNA and protein levels. RESULTS AND CONCLUSION: Under the scanning electron microscope, MG63 cells adhered well to the scaffold surface. Compared with the blank group, the proliferation of MG63 cells could be significantly promoted by either platelet lysate or porous tantalum scaffold (P < 0.05). Moreover, the proliferation of MG63 cells was significantly improved in the combined group compared with the other three groups (P < 0.05). Findings from qPCR, immunocytochemical staining and western blot showed the highest expression of ITGβ1, Vinculin, F-actin mRNA and protein in the combined group (P < 0.05). These results indicate that platelet lysate and the domestic porous tantalum scaffold can synergistically promote the proliferation of MG63 cells, and up-regulate the expression of ITGβ1, Vinculin and F-actin mRNA and protein. Activation of the ITGβ1/Vinculin/F-actin signaling pathway may contribute to the proliferation, adhesion and differentiation of MG63 cells.

OBJECTIVETo identify gene mutations of a pedigree with inherited factor V (FV) deficiency.

METHODSThe activated partial thromboplastin time (APTT), prothrombin time (PT), FV activity (FV:C) and FV antigen (FV:Ag) tests were performed for phenotypic diagnosis. The genomic DNA was extracted from the peripheral blood of the proband and all the 25 exons and their flanks of FV gene were amplified by polymerase chain reaction (PCR). The PCR products were screened by direct sequencing and the mutations were further confirmed by restriction enzyme digestion.

RESULTSAPTT, PT, TT, FV:C, FV:Ag of the proband were 249.2 s, 46.6 s, 17.9 s, 0.1% and 1.5%, respectively. FII, FVII, FVIII, FIX, FX activities, vWF and Fg were within normal ranges. Taking the GenBank Z99572 sequence as the reference, four mutations were identified in FV gene of the proband. They were a heterozygous two bases deletion in exon 13 (2238 approximately 2239delAG) introducing a frameshift and a premature stop at codon 689, and a heterozygous missense mutation in exon 23 (G6410T) resulting in the substitution of Gly for Val at codon 2079, respectively. The proband's father and mother were heterozygous for G6410T and for 2238 approximately 2239delAG, respectively.

CONCLUSIONThe severe FV deficiency of the proband is caused by a frameshift mutation of 2238 approximately 2239delAG and a missense mutation of G6410T, which haven't been identified before.

Adult ; Base Sequence ; DNA Mutational Analysis ; Exons ; genetics ; Factor V ; genetics ; metabolism ; Factor V Deficiency ; genetics ; Female ; Frameshift Mutation ; Heterozygote ; Humans ; Infant ; Male ; Mutation, Missense ; Partial Thromboplastin Time ; Pedigree ; Phenotype ; Prothrombin Time ; Thrombin Time

To investigate the clinical features and prognostic significance in myelodysplastic syndrome (MDS) patients with DTA (DNMT3A,TET2,ASXL1) mutations. Clinical characteristics of 140 patients diagnosed as de novo MDS at People′s Hospital of Xinjiang Uygur Autonomous Region from September 2015 to December 2019 were retrospectively analyzed. Next-generation sequencing was used to detect 34 related genes in MDS patients. DTA mutations and the correlation with progression-free survival (PFS) and overall survival (OS) in MDS patients were evaluated. Among 140 MDS patients, DTA mutations was detected in 62 (44.3%) patients. And the positive rate of DTA mutations in IPSS-R lower-risk group was 65.4%, significantly higher than that of higher-risk group (31.8%)( P=0.000). Compared with the non-mutated group, patients with DTA mutations had a lower rate of conversion to leukemia (9.7% vs . 29.5%, P=0.004).Survival analysis showed that PFS in patients with DTA mutations was comparable as that in MDS patients without DTA mutations ( P=0.787), but the median OS was significantly shorter (16 months vs . 20 months, P=0.022).According to IPSS-R classification, the median OS in patients with and without DTA mutation was only statistically significant in the higher-risk group (15 months vs. 18 months, P=0.034).Among 62 patients with DTA mutations, 60 (96.8%) had additional gene mutations. DTA mutations were not independent prognostic factors when mutation frequency is greater than 10% were considered in Cox regression model ( P>0.05). DTA mutations often developed in the early stage of MDS, therefore they were more common in IPSS-R lower-risk subgroup which was correlated to the low rate of conversion to leukemia. In conclusion, DTA mutations are not associated with disease progression, but predict unfavorable survival when other add-on genes are mutated.

OBJECTIVE: To investigate the hypothesis that hydrogen could ameliorate cecal ligation and puncture (CLP)-induced lung injury of rats by inhibiting cystathionine-gamma-lyase/hydrogen sulfide (CSE/HS) system. METHODS: A total number of 24 healthy male SD rats weighting 250～300 g were randomly divided into four groups (n=6 in each group): sham operation group(sham group), hydrogen-rich saline control group(H group), CLP group and hydrogen-rich saline treatment group(CLP+H group). The rats were treated with hydrogen-rich saline or saline 10 min before CLP or sham operation. At 8 h of sham or CLP operation, lung samples were obtained to detect the changes of the CSE/HS system using biochemical and RT-PCR methods. In order to further confirm the role of HS during hydrogen improve the lung injury of CLP rats, we also observed the effect of hydrogen-rich saline on the lung injury induced by HS donor-sodium sodium hydrosulfide (NaHS). Thirty-two healthy male SD rats (250~300 g) were randomly divided into four groups (n=8 in each group): control group, HS group, HS+H group and H group. Saline(10 mg/kg) or NaHS(HS donor, 56 μmol/kg) was injected intraperitoneally (10 mg/kg) respectively into rats in the control rats or HS group. For rats in the HS+H and H group, hydrogen-rich saline (10 mg/kg) was injected 10 min before saline or NaHS administration. Eight hours after the LPS saline or NaHS administration, lung coefficient, MDA content, and MPO activity were detected. The contents of TNF-α, IL-6 and IL-10 in lung tissue were measured, and the morphological changes of lung tissue were also observed. RESULTS: CSE/HS system up-regulating were observed in animals exposed to CLP. Hydrogen-rich saline treatment significantly inhibited CSE/HS system as indicated by significantly reduced HS production in lung, along with a decreased CSE activity and CSE mRNA expression (all P＜0.05). Importantly, the results showed that lung injury and lung tissue inflammation were observed in animals exposed to NaHS. Hydrogen-rich saline treatment significantly attenuated lung injury as indicated by significantly improved histological changes in lung, significantly reduced index of quantitative assessment (IQA), MDA content and lung coefficient (all P＜0.05). MPO activity in lung tissue was significantly reduced along with decreased productions of TNF-α and IL-6, and an increased production of IL-10 in the presence of hydrogen (all P＜0.05), demonstrating antioxidant and anti-inflammatory effect of hydrogen in NaHS-induced ALI. CONCLUSION These results indicate that hydrogen-rich saline peritoneal injection improves the lung injury induced by CLP operation. The therapeutic effects of hydrogen-rich saline may be related to suppressing the production of HS.
Animals ; Cecum ; surgery ; Cystathionine gamma-Lyase ; metabolism ; Cytokines ; metabolism ; Hydrogen ; pharmacology ; Hydrogen Sulfide ; metabolism ; Ligation ; Lung Injury ; therapy ; Male ; Punctures ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Saline Solution ; pharmacology