OBJECTIVETo identify the genetic defect underlying congenital afibrinogenemia in a Chinese family.

METHODSPlasma fibrinogen (Fg) was assessed by both Clauss method and immunonephelometry. Genomic DNA was isolated from peripheral blood of the proband and 13 members of her family. All the exons and exon-intron boundaries of the three fibrinogen genes (FGA, FGB, FGG) were amplified by PCR followed by direct sequencing. Restriction endonuclease analysis was performed for the PCR products of the family members and 50 healthy donors to exclude gene polymorphism.

RESULTSNo Fg was detected in the plasma of the proband and her father by Clauss method, while low levels (< 0.02 g/L) were detected by immunonephelometry. A homozygous C to T mutation was found in the two cases at nucleotide 3108 in exon 4 of FGA gene, resulting in a null mutation which encoded severely truncated alpha-chains owing to its premature termination at the Gln 150 codon. The C-->T mutation eliminated a unique recognition site for restriction enzyme RsaI. The PCR amplified fragments of the proband and her father could not be digested by RsaI, showing that they are homozygous. Her mother and some family members are heterozygous at this site since the fragment could partly be digested, while the same fragment of controls could be completely digested as expected.

CONCLUSIONThe Gln (CAG)-->150stop (TAG) nonsense mutation in FGA gene is a novel genetic defect of congenital afibrinogenemia which, to our knowledge, has not been described before.

Adolescent ; Afibrinogenemia ; congenital ; genetics ; Base Sequence ; Codon, Nonsense ; DNA Mutational Analysis ; Exons ; genetics ; Female ; Fibrinogen ; genetics ; Humans ; Male ; Pedigree

OBJECTIVETo observe the dynamic expression of calcium-sensing receptor(CaSR) in myocardium of diabetic rats.

METHODSThirty male Wistar rats were randomly divided into 3 groups including control, diabetic-4 week and diabetic-8 week groups(n = 10). The type 2 diabetes mellitus models were established by intraperitoneal injection of streptozotocin (STZ, 30 mg/kg) after high-fat and high-sugar diet for one month. The cardiac morphology was observed by electron microscope. Western blot analyzed the expression of CaSR, phospholamban (PLN), a calcium handling regulator, and Ca+-ATPase(SERCA) in cardiac tissues.

RESULTSCompared with control group, the expressions of CaSR and SERCA were decreased, while the expression of PLN was significantly increased in a time-dependent manner in diabetic groups. Meanwhile diabetic rats displayed abnormal cardiac structure.

CONCLUSIONThese results indicate that the CaSR expression of myocardium is reduced in the progression of DCM, and its potential mechanism may be related to the imnaired intracellular calcium homeostasis.

Animals ; Calcium-Binding Proteins ; metabolism ; Diabetes Mellitus, Experimental ; complications ; Diabetes Mellitus, Type 2 ; Diabetic Cardiomyopathies ; metabolism ; physiopathology ; Disease Progression ; Heart ; physiopathology ; Male ; Myocardium ; metabolism ; pathology ; Rats ; Rats, Wistar ; Receptors, Calcium-Sensing ; metabolism ; Sarcoplasmic Reticulum Calcium-Transporting ATPases ; metabolism ; Streptozocin

OBJECTIVETo identify the genetic defect of inherited coagulation factor (F) deficiency in a Chinese family and to explore its molecular mechanism.

METHODSThe activity and antigen of plasma F were measured by photometric test and enzyme-linked immunosorbent assay, and rocket-electrophoresis, respectively. All the exons and exon-intron boundaries of the FA subunit gene were amplified by PCR and then DNA sequencing was performed. Restriction endonuclease analysis was used for the PCR products of the family members and 80 healthy donors to exclude gene polymorphism.

RESULTSRapid dissolution of the proband's fibrin clot occurred within 30 minutes, and antigen of his plasma F was significantly decreased, two compound heterozygous missense mutations (a C to T transition at nucleotide 177,246 which caused Arg703Trp, and a A to G transition at nucleotide 177,286 which caused His716Arg) in exon 15 of FA subunit gene were found. The possibility of gene polymorphism was excluded by restriction endonuclease analysing. Each of these two missense mutations was respectively found in his mother and father. Molecular modeling based on 3D crystallographic data predicted that the mutant protein decreased stability and was likely to be rapidly degraded.

CONCLUSIONSThe inherited F deficiency in the Chinese family is caused by two compound heterozygous missense mutations-Arg703Trp and His716Arg in the FA subunit, which to our knowledge, are reported for the first time.

Base Sequence ; Child ; Exons ; Factor XIII ; genetics ; Factor XIII Deficiency ; genetics ; Heterozygote ; Humans ; Male ; Molecular Sequence Data ; Mutation, Missense ; Pedigree

OBJECTIVETo investigate the effects of losartan on left ventricular hypertrophy (LVH) and plasma transforming growth factor-beta1 (TGF-beta1) in elderly patients with essential hypertension (EH).

METHODSThe elderly patients with EH were divided into two groups, namely EH+LVH group and EH group according to the data of echocardiogram. The systolic and diastolic blood pressures of the patients were monitored. Plasma TGF-beta1 was measured before and after 6 months' treatment with losartan, and the relationship between TGF-beta1 and other index were analyzed.

RESULTSAfter 6 months' treatment, the blood pressure of EH+LVH group and EH group were significantly lowered (P<0.01). Significant improvement of IVSTd, LVPWd, E/A, and LVMI (P<0.01) and obvious reduction of plasma TGF-beta1 (P<0.01) occurred in EH+LVH group after 6 months' treatment. Correlation analyses indicated that the plasma TGF-beta1 level was positively correlated to LVMI (P<0.01).

CONCLUSIONLosartan can reversed LVH in elderly patients with EH partially by lowering plasma TGF-beta1 level.

Aged ; Antihypertensive Agents ; therapeutic use ; Female ; Humans ; Hypertension ; blood ; complications ; drug therapy ; Hypertrophy, Left Ventricular ; blood ; drug therapy ; etiology ; Losartan ; therapeutic use ; Male ; Middle Aged ; Transforming Growth Factor beta1 ; blood

OBJECTIVETo assess the diagnostic value of 8 equations using different variables for determining the estimated glomerular filtration rate (eGFR) in patients with cardiovascular diseases.

METHODSGFR was estimated in 208 patients with cardiovascular diseases by (99m)Tc-DTPA dynamic renal imaging, and the eGFR was derived from 8 equations using different variables.

RESULTSIn patients with chronic kidney disease (CKD) stages 1-3, the eGFR calculated suing serum creatinine (SCr)-based equation was better correlated to GFR estimated by (99m)Tc-DTPA renal imaging than that derived from cystatin C (Cys C)-based equations, whereas in patients with CKD stages 4 and 5, the estimates by the latter equation showed a better correlation to GFR. Compared with (99m)Tc-DTPA renal imaging, MDRD-based equation and simple MDRD equation resulted in a higher eGFR in patients with CKD stages 4 and 5, the Rule equation had a lower eGFR in CKD stages 1 and 2, the Macisaac equation yielded a higher eGFR in CKD stages 2-5, and the Tan equation showed a higher eGFR in CKD stages 2 and 3. In patients with mild renal dysfunction, the Scr-based equation had a higher AUC(ROC) than Cys C-based equation, which was reversed in patients with severe renal dysfunction; the AUC(ROC) of the two equations were comparable in patients with moderate renal dysfunction. Compared with (99m)Tc-DTPA renal imaging, the modified MDRD equation and Arnal-Dade equation showed no significant difference in the eGFR in patients with CKD stages 1-5.

CONCLUSIONModified MDRD equation (or simple MDRD equation) and Arnal-Dade equation are superior to other calculation methods for estimating the GFR in Chinese patients with cardiovascular disease.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cardiovascular Diseases ; physiopathology ; Child ; Chronic Disease ; Creatinine ; blood ; Cystatin C ; blood ; Female ; Glomerular Filtration Rate ; physiology ; Humans ; Kidney Diseases ; physiopathology ; Male ; Middle Aged ; Young Adult

OBJECTIVETo evaluate the efficacy and safety of irinotecan combined with xeloda (CAPIRI regimen) in patients with metastatic colorectal cancer after failure of chemotherapy with oxaliplatin.

METHODSTotally 38 patients with metastatic colorectal cancer after failure of chemotherapy with oxaliplatin were enrolled. Patients received xeloda 1 000 mg/m2 orally twice daily on day 1 to 14 and intravenous irinotecan 100 mg/m2 on day 1 and 8 every 3 weeks.

RESULTSThe median age of 38 patients was 58.5 (27-77) years. CAPIRI regimen was used 11.0 (3.0-24.0) months after the diagnosis of metastatic colorectal cancer (CAPIRI regimen as second-line chemotherapy in 33 patients, third-line in 4 patients, and fourth-line in 1 patient). A total of 121 cycles of chemotherapy (median 3.0) were administered. Thirty-four patients were evaluable for response. The overall response rate and disease control rate were 5.9% and 61.8%, respectively. The median time to progression and overall survival were 4.5 months (95% CI, 3.4-5.6 months) and 11.0 months (95% CI, 10.2-11.8 months), respectively. All 38 patients were evaluable for safety. The most common adverse events were leukopenia (73.7%), neutropenia (65.8%), nausea and vomiting (60.5%), and diarrhea (28.9%). The occurrence rates of these grade 3-4 events were 10.5%, 13.2%, 10.5%, and 7.9%, respectively. All adverse events were tolerable.

CONCLUSIONCAPIRI regimen is effective and well-tolerated in Chinese patients with metastatic colorectal cancer after failure of chemotherapy with oxaliplatin.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Camptothecin ; administration & dosage ; analogs & derivatives ; Capecitabine ; Colorectal Neoplasms ; drug therapy ; Deoxycytidine ; administration & dosage ; analogs & derivatives ; Female ; Fluorouracil ; administration & dosage ; analogs & derivatives ; Humans ; Male ; Middle Aged ; Neoplasm Metastasis ; Organoplatinum Compounds ; therapeutic use ; Treatment Outcome

BACKGROUNDGrowing preclinical evidence shows that zoledronic acid (ZOL) exhibits direct antitumor activity in various cancer cell lines. However, the cytotoxic effects of ZOL on human hepatocellular carcinoma (HCC) cells have not been established. In the present study, we investigated the effect of ZOL on HCC both in vitro and in vivo.

METHODSCytotoxicity and cell cycles were assessed with Sulforhodamine B colorimetric assay and flow cytometry. Expression levels of cell cycle phase-linked proteins were examined. The effect of ZOL on HCC in vivo was explored based on H22-subcutaneous injection (s.c.) and H22-intraperitoneal injection (i.p.) mice model.

RESULTSZOL inhibited the growth of SK-HEP-1 and H22 cells and induced S-phase arrest through downregulating cdc2 protein and upregulating cyclin A. It inhibited the growth of s.c tumors, and increased the survival of both H22-s.c. and H22-i.p. mice in vivo.

CONCLUSIONZOL inhibits growth of HCC cells in vitro and in vivo.

Animals ; Carcinoma, Hepatocellular ; drug therapy ; pathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Diphosphonates ; pharmacology ; therapeutic use ; Female ; Humans ; Imidazoles ; pharmacology ; therapeutic use ; Liver Neoplasms ; drug therapy ; pathology ; Mice ; Xenograft Model Antitumor Assays

OBJECTIVETo compare the differences in the efficacy on distant version of naked eye in the patients of juvenile myopia between rotating manipulation and lifting-thrusting manipulation of acupuncture.

METHODSOne hundred and twenty cases (240 eyes) were randomized into a rotating manipulation group and a lifting-thrusting manipulation group, 60 cases (120 eyes) in each group. Additionally, a corrective lenses group, 60 cases (120 eyes), was set up as the control. In both manipulation groups, Cuanzhu (BL 2),Yuyao (EX-HN 4), Sizhukong (TE 23), Taiyang (EX-HN 5), Fengchi (GB 20), Zusanli (ST 36), Guangming (GB 37) and Sanyinjiao (SP 6) were punctured, but stimulated with rotating manipulation and lifting-thrusting manipulation respectively three times per week, 10 times as a treatment session and totally one session was required. In the corrective lenses group, the glasses were applied at daytime. The clinical efficacy and the changes in distant vision of naked eye before and after treatment were compared among the three groups.

RESULTSThe total effective rate was 87.5% (105/120) in the rotating manipulation group, which was better than 69.2% (83/120) in the lifting-thrusting manipulation group (P < 0.05). The distant vision of naked eye was improved apparently in the rotating manipulation group and the lifting-thrusting manipulation group after treatment (both P < 0.05). But it was not improved in the corrective lenses group (P > 0.05). The distant vision of naked eye was improved more apparently after treatment in the rotating manipulation group as compared with that in the lifting-thrusting manipulation group (0.75 +/- 0.23 vs 0.68 +/- 0.24, P < 0.05). For 96 cases (192 eyes) with acupuncture treatment, in 3-month follow-up, 87.0% (167/192) of the cases maintained the stable vision as the original level and 13.0% (25/192) of them were reduced in the vision In the acupuncture groups, it was found that the improvement of distant vision of naked eye was more obvious after treatment with younger age, better basic vision and shorter duration of sickness (all P < 0.05).

CONCLUSIONAcupuncture achieves the positive and sustainable clinical effect on juvenile myopia, and the results of rotating manipulation are superior to that of lifting-thrusting manipulation. Age, basic vision and duration of sickness impact the clinical efficacy.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; methods ; Adolescent ; Child ; Female ; Humans ; Myopia ; therapy ; Young Adult

OBJECTIVETo summarize the clinical pathological characteristics and treatment patterns of breast cancer in elderly women.

METHODSA total of 87 patients (≥ 60 years) admitted to our hospital between January and December 2007 were included in this retrospective study. The patients were divided into 60-69-year group and ≥ 70-year group, and their clinical pathological data and treatment modes were summarized and compared.

RESULTSThe tumor size (T2-T3), number of involved axillary lymph nodes,and positive rates of estrogen/progesterone receptors,over-expression of epidermal growth factor receptor 2, and ≥ 2 complication were not significantly different between two groups (P > 0.05). The ≥ 70-year group tended to have similar p53 gene mutation and Ki-67 labeling index with the 60-69-year group, although the P values were close to 0.05 (P = 0.09, P = 0.08,respectively). In the ≥ 70-year group, 33.3% of patients underwent extended resection,while in the 60-69-year group, all patients received modified radical treatment (P < 0.005). The percentages of adjuvant chemotherapy were 25% and 56.9% in the ≥ 70-year group and the 60-69-year group (0.005). The percentages of adjuvant endocrine therapy applied after surgery were similar in 2 groups (77.8% and 68.6% separately, P=0.347). Binary logistic regression showed that age,number of involved axillary lymph nodes,and estrogen receptor-positive rate were independently associated with adjuvant chemotherapy,while the pathological tumor size and complication were irrelevant. The 2-year disease-free survival rates of 2 groups were not significantly different.

CONCLUSIONSThe clinical pathological characteristics of breast cancer were similar in elderly patients who are 60-69 years old or ≥ 70 years. In the treatment pattern,patients who are ≥ 70 years tend to receive endocrine therapy rather than adjuvant chemotherapy.

Aged ; Aged, 80 and over ; Breast Neoplasms ; pathology ; therapy ; Female ; Humans ; Middle Aged ; Retrospective Studies

OBJECTIVETo observe the effect of dopamine receptor (DR2) activation on hypoxia/reperfusion injury (HRI) in the neonatal rat cardiomyocytes, and to explore its mechanism.

METHODSThe hypoxia/reperfusion (H/R) injury model was established in primarily cultured neonatal rat cardiomyocytes, and randomly assigned: control, H/R, bromocriptine (Bro) and haloperidol (Hal) groups. The cell apoptosis was detected using inverted microscope, transmission electron microscope and flow cytometry (FCM). The lactate dehydrogenase(LDH) and superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in cell medium were analyzed. The expression of mRNA and protein of caspase-3, caspase-8, caspase-9, Fas, Fas-L, Cyt C and Bcl-2 were detected by RT-PCR and Western blot, respectively.

RESULTSCompared with the control group, apoptosis rate, LDH activity, MDA content and the expression of pro-apoptotic factors and anti-apoptotic factors were increased, but SOD activity was decreased in H/R group. Compared with the H/R group, all index above-mentioned were down-regulated or reversed in Bro-group, and had no obvious differences in Hal-group.

CONCLUSIONThe neonatal rat cardiomyocytes injury and apoptosis caused by hypoxia/reperfusion can be inhibited with DR2 activation, which mechanism is related to scavenging oxygen radical.

Animals ; Animals, Newborn ; Apoptosis ; Cell Hypoxia ; Myocardial Reperfusion Injury ; etiology ; metabolism ; Myocytes, Cardiac ; cytology ; metabolism ; Oxidative Stress ; Rats ; Rats, Wistar ; Receptors, Dopamine D2 ; metabolism