Isaridin E Protects against Sepsis by Inhibiting Von Willebrand Factor-Induced Endothelial Hyperpermeability and Platelet-Endothelium Interaction.

Endothelial hyperpermeability is pivotal in sepsis-associated multi-organ dysfunction. Increased von Willebrand factor (vWF) plasma levels, stemming from activated platelets and endothelium injury during sepsis, can bind to integrin αvß3, exacerbating endothelial permeability. Hence, targeting this pathway presents a potential therapeutic avenue for sepsis. Recently, we identified isaridin E (ISE), a marine-derived fungal cyclohexadepsipeptide, as a promising antiplatelet and antithrombotic agent with a low bleeding risk. ISE's influence on septic mortality and sepsis-induced lung injury in a mouse model of sepsis, induced by caecal ligation and puncture, is investigated in this study. ISE dose-dependently improved survival rates, mitigating lung injury, thrombocytopenia, pulmonary endothelial permeability, and vascular inflammation in the mouse model. ISE markedly curtailed vWF release from activated platelets in septic mice by suppressing vesicle-associated membrane protein 8 and soluble N-ethylmaleide-sensitive factor attachment protein 23 overexpression. Moreover, ISE inhibited healthy human platelet adhesion to cultured lipopolysaccharide (LPS)-stimulated human umbilical vein endothelial cells (HUVECs), thereby significantly decreasing vWF secretion and endothelial hyperpermeability. Using cilengitide, a selective integrin αvß3 inhibitor, it was found that ISE can improve endothelial hyperpermeability by inhibiting vWF binding to αvß3. Activation of the integrin αvß3-FAK/Src pathway likely underlies vWF-induced endothelial dysfunction in sepsis. In conclusion, ISE protects against sepsis by inhibiting endothelial hyperpermeability and platelet-endothelium interactions.

Antiplatelet and Antithrombotic Effects of Isaridin E Isolated from the Marine-Derived Fungus via Downregulating the PI3K/Akt Signaling Pathway.

Isaridin E, a cyclodepsipeptide isolated from the marine-derived fungus Amphichorda felina (syn. Beauveria felina) SYSU-MS7908, has been demonstrated to possess anti-inflammatory and insecticidal activities. Here, we first found that isaridin E concentration-dependently inhibited ADP-induced platelet aggregation, activation, and secretion in vitro, but did not affect collagen- or thrombin-induced platelet aggregation. Furthermore, isaridin E dose-dependently reduced thrombosis formation in an FeCl3-induced mouse carotid model without increasing the bleeding time. Mechanistically, isaridin E significantly decreased the ADP-mediated phosphorylation of PI3K and Akt. In conclusion, these results suggest that isaridin E exerts potent antithrombotic effects in vivo without increasing the risk of bleeding, which may be due to its important role in inhibiting ADP-induced platelet activation, secretion and aggregation via the PI3K/Akt pathways.

Predictive and prognostic values of preoperative platelet parameters in patients with gynecological tumors.

BACKGROUND: Platelets play a role in tumor cell growth, metastasis, and angiogenesis, and the present study aimed to evaluate diagnostic and prognostic values of platelet parameters in patients with gynecological tumors. METHODS: A total of 1062 women were included. Differences of platelet parameters (platelet count [PLT], plateletcrit [PCT], mean platelet volume [MPV], platelet-large cell rate [P-LCR], and platelet distribution width [PDW]) between different categories were analyzed by nonparametric test. The optimal cutoff value was calculated with receiver operating characteristic analysis. Overall survivals were analyzed with Kaplan-Meier method and log-rank tests for univariate analysis. RESULTS: Platelet count and PCT were significantly increased, and MPV and P-LCR were significantly reduced in malign and benign gynecological tumor groups compared with the controls (P < .001); PDW had no significant differences. There were no significant differences in PLT, PCT, MPV, P-LCR, and PDW between different tumor locations and pathologic types. The optimal cutoff values of PLT, PCT, MPV and P-LCR were 274, 0.26, 10.08, and 24.8 (AUC: 0.661, 0.643, 0.593, 0.562), and PCT had preferable sensibility and specificity (50.84% and 70.42%) in predicting the presence of gynecological tumors. According to survival analysis, increased PLT (≥274 × 109 /L) and PCT (≥0.26), and induced MPV (<10.08 fL) and P-LCR (<24.8%) were associated with shorter overall survival. CONCLUSIONS: Platelet count, PCT, MPV, and P-LCR can be used as preferable auxiliary parameters for predicting the presence of gynecological tumors. Increased PLT and PCT, or decreased MPV and P-LCR indicated a heavier tumor burden and shorter overall survival.

Correlation between bone marrow dose volumes and acute hematological toxicity in postoperative gynecological cancer patients.

OBJECTIVE: To identify the association between radiation dose volume and acute hematological toxicity (HT) in postoperative gynecological cancer patients receiving whole pelvic radiotherapy (RT) or intensity-modulated RT (IMRT), a principal component regression model was used to calculate HT. METHODS: Women (n=100) receiving with or without chemotherapy RT were retrospectively analyzed, 52 of whom received chemotherapy (paclitaxel and nedaplatin). The pelvis and lumbar vertebrae, defined as the prolong-pelvic bone marrow, were divided into the (1) combined ilium, ischium and pubis and the (2) lumbar vertebrae and the sacrum. The V5-V40 of subsides was calculated. The complete blood counts were recorded weekly. The principal component analysis was performed on volumes which generated the principal components (PCs), followed by using a logistic regression model. RESULTS: Forty-seven patients presented with grade 2-3 HT during RT. Chemotherapy increased the incidence of HT compared with RT alone (70.21% vs. 29.79%; p=0.001). Fifty-three patients with persistent HT developed more serious HT at an earlier stage of RT. The chemotherapy cycles and three PCs associated with grade 2-3 HT was identified to form the resulting principal logistic regression model. CONCLUSION: A new method to calculate the NTCP was achieved by PCs logistic regression.

[Identification and functional analysis of a novel missense mutation Ser250Phe underlying congenital coagulation factor â ¦ deficiency].

OBJECTIVE: To identify and characterize a missence mutation Ser250 Phe underlying coagulation factor Ⅶ (FⅦ) deficiency in a Chinese patient and his family. METHODS: The FⅦ gene (F7) was analyzed by DNA sequencing, and the FⅦ levels (including antigen and activity) in patient's plasma were determined with enzyme-linked immunoabsorbent assay (ELISA) and one stage prothrombin time based method. In addition, a FⅦ-250 Phe mutant corresponding to the identified mutation was expressed in HEK293 cells, and a subcellular localization experiment in CHO cells was performed to clarify the molecular mechanism of FⅦ deficiency caused by the FⅦ-250 Phe mutation. RESULTS: The patient had a prolonged prothrombin time (PT: 36.5 s) and low levels of both FⅦ antigen and activity (130.2 ng/mL and 4.0%, respectively). Two heterozygous mutations were identified in the F7 gene (NG-009262.1), which included a g.15975 G>A mutation at the splice receptor site of intron 6 (IVS6-1G>A) and a novel g.16750 C>T mutation in exon 8, which resulted in replacement of Ser (TCC) 250 with Phe (TTC)250 in the vicinity of a charge-stablizing system. By gene expression experiments, the antigen and activity levels of FⅦ-250 Ser and FⅦ-250 Phe in the culture medium were (37.77 ± 2.30) ng/mL and (4.02 ± 0.52) ng/mL, respectively. ELISA and Western blotting analyses indicated that expression of the mutant FⅦ-250 Phe and wild type FⅦ-250 Ser was (130.51 ± 2.32) ng/mL and (172.45 ± 2.25) ng/mL, respectively. FⅦ-250 Phe was found in endoplasmic reticulum and Golgi apparatus, suggesting that the mutant FⅦ-250 Phe could be normally synthesized in the cells but was inefficiently secreted. CONCLUSION: Compound heterozygous mutations in F7 gene (g.15975G>A and g.16750C>T) may be responsible for the FⅦ deficiency in this patient. The novel FⅦ 250 Phe can be transported from endoplasmic reticulum to Golgi apparatus, but may be degraded or inefficient.

[Dexketoprofen trometamol in the treatment of chronic prostatitis/chronic pelvic pain syndrome].

OBJECTIVE: To evaluate the clinical efficacy and safety of dexketoprofen trometamol in the treatment of patients with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS). METHODS: A total of 115 patients with CP/CPPS were divided into a dexketoprofen trometamol group (n = 40), treated with dexketoprofen trometamol (25 mg, tid) and terazosin (2 mg, qn), an indometacin group (n = 40) given indometacin (25 mg, tid) and terazosin (2 mg, qn), and a terazosin group (n = 35) administered terazosin (2 mg, qn) only, all treated for 4 weeks. Scores on the NIH-chronic prostatitis symptom index (NIH-CPSI) were obtained before and after the treatment, and the efficacy and adverse events were observed and compared. RESULTS: The NIH-CPSI scores were significantly improved after the treatment in all the three groups. The clinical efficacy was significantly better in the dexketoprofen trometamol and indometacin groups than in the terazosin group (P < 0.05), but with no significant difference between the former two (P > 0.05). The rates of adverse events were 10.00%, 18.57% and 27.50% in the dexketoprofen trometamol, terazosin and indometacin groups, significantly lower in the former two than in the latter one (P < 0.05). CONCLUSION: The combination of dexketoprofen trometamol with terazosin could effectively improve the clinical symptoms of CP/CPPS, better than terazosin in therapeutic efficacy and than indometacin in drug tolerance.

[Genetic Diagnosis and Phenotype Analysis for 3 Patients with Hereditary Coagulation Factor â ¦ Deficiency].

OBJECTIVE: To investigate the gene mutations types and the clinical characteristics in 3 patients with hereditary coagulation factor Ⅶ deficiency. METHODS: The phenotype diagnosis was validated by detecting the coagulation parameters including prothrombin time (PT)，activated partial thromboplastin time (APTT), fibrinogen (FIB), FⅦ activity (FⅦ: C) and specific antigens (FⅦ: Ag) of proband and its family members. All exons, exon-intron boundaries, 5＇ untranslated regions and 3＇ untranslated regions of F7 gene were amplified with PCR. Potential mutations were detected by direct sequencing of purified PCR products. Suspected mutations were confirmed by sequencing of the opposite strand. RESULTS: A total of 5 different mutations were identified in 3 patients with hereditary coagulation factor Ⅶ deficiency and family members, including 4 misssense mutations and 1 splice site mutation. Out of 3 cases of hereditary coagulation factor Ⅶ deficiency 2 had double heterozygous mutation, I had homozygous mutations. Patient 1 had p.His408Gln with p.Arg413Gln double heterozygous mutations, her sister had p.His408Gln with p.Arg413Gln double heterozygous mutations, another one had p.His408Gln mono-heterozygous mutation, their correspo FⅦ: C were 5%, 3%, 75%. Patient 2 had p.Arg364Gln with p.His408Gln double heterozygous mutations, her brother had p.Arg364Gln with IVS6-1G＞A double heterozygous mutations, their corresponding FⅦ: C were 2.0%, 2.0%. Patient 3 had p.Arg337Cys homozygous mutation, FⅦ: C was 3.0%. CONCLUSION: A total of 5 different mutations were identified in 3 patients with hereditary coagulation factor Ⅶ deficiency, the p.His408Gln is a common mutation, the FⅦ: C and FⅦ: Ag have no correlation with clinical phenotypes.

[Effect of different blood glucose levels on the expression of cerebral GLUT3 mRNA in neonatal rats with hypoxia-ischemia].

OBJECTIVE: Concerns of the effect of glucose on perinatal hypoxic-ischemic brain damage are increasing. It was previously considered that the glucose transporter (GLUT) genes and their productions played an important role in the regulation of cerebral energy metabolism. The present study aimed to explore the effect of different blood glucose levels on the expression of cerebral GLUT3 mRNA in neonatal rats with hypoxia-ischemia (HI), and to evaluate the neuroprotective effect of glucose against HI insults. METHODS: A total of 250 7-day-old neonatal SD rats were randomly divided into 10 groups (n=25 each): Normal control, Sham-operated, HI, Hypoglycemia, Hypoglycemia pre- and post-HI, Mild hyperglycemia pre- and post-HI, Severe hyperglycemia pre- and post-HI. Blood glucose levels of normal, hypoglycemia, mild hyperglycemia and severe hyperglycemia were defined as 5-7 mmol/L, 3-4 mmol/L, 10-15 mmol/L and 16-25 mmol/L, respectively. The expression of GLUT3 mRNA was detected with RT-PCT technique at 2, 24, 48 and 72 hrs and at 7 days after HI. RESULTS: There was a correlation between increases in GLUT3 mRNA expression and postnatal age in the Normal control group. HI significantly enhanced the expression of GLUT3 mRNA from 2 hrs, peaking at 24 hrs after HI, and then significantly decreased at 72 hrs and 7 days after HI when compared with the Normal Control group (P < 0.01). GLUT3 mRNA expression in the Hypoglycemia pre-HI group was the lowest among all groups with HI at each time point after HI, and a statistically significant difference was found at 72 hrs after HI when compared with the HI group (P < 0.05). The expressional levels of GLUT3 mRNA in the Severe hyperglycemia pre-HI group were strikingly higher than those in any other groups with HI (P < 0.05 or 0.01). The GLUT3 mRNA expression patterns in the Mild and Severe hyperglycemia post-HI and the Hypoglycemia post-HI groups were similar to the Hypoglycemia pre-HI group. CONCLUSIONS: GLUT3 mRNA expression and the synthesis of GLUT3 can be down-regulated by hypoglycemia pre-HI, coupled with aggravation of cerebral pathology, but up-regulated by higher hyperglycemia pre-HI, coupled with improvement of cerebral pathology. This suggested that adequate glucose supplement before HI can improve the cerebral function against HI insults in neonatal rats.

Hepatoprotective function of Penthorum chinense Pursh.

This research was to figure out the hepatoprotective constituents of Penthorum chinense Pursh, a typical species both for food and medicine, using carbon tetrachloride-induced chronic hepatotoxicity in HL-7702 cells. Cell viability, levels of microsomal enzymes, alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and the concentration of malondialdehyde (MDA) were determined in homogenates of the liver cells. Results showed that the cell viability increased significantly in all test groups in a concentration-dependent manner and the content of the peroxidation product MDA decreased significantly, similar to the serum levels of hepatic enzyme biomarkers (ALT and AST). The effects of 70% ethanol extracts and Vc were better than 95% ethanol and water extracts, and the ethyl acetate extracts further obtained from 70% ethanol fraction showed the highest hepatoprotective activity, even better than Vc. The ethyl acetate fraction from 70% ethanol extracts is responsible for the hepatoprotective function of P. chinense.

Antioxidant and anticomplement functions of flavonoids extracted from Penthorum chinense Pursh.

Penthorum chinense Pursh is rich in flavonoids, which have strong antioxidant and anticomplement activities. In order to optimize their extraction conditions, various extraction parameters were chosen to identify their effects on flavonoids extraction. Single factor and Box-Behnken experimental designs consisting of 24 experimental runs and five replicates at zero point were applied to obtain the optimal extraction yield. The optimization conditions for flavonoids extraction were determined as follows: ethanol concentration 60.89%, extraction time 68.15 min, temperature 52.89 °C and liquid/solid ratio 19.70 : 1. The corresponding flavonoids content was 7.19%. The regression equation was found to fit well with the actual situation. Furthermore, the antioxidant activity (the free radical scavenging ability and ferric reducing/antioxidant power) and anticomplement ability of the flavonoids from P. chinense were determined. Results showed that the flavonoids of P. chinense displayed significant antioxidant and anticomplement activities. Its antioxidant activity can compete with ascorbic acid (Vc), whereas its anticomplement activity (IC50 = 111.6 µg ml(-1)) surpassed the effect of heparin (IC50 = 399.7 µg ml(-1)) which was used as the positive control, suggesting that P. chinense flavonoids and their related products could potentially be used as a promising natural agent in the treatment of humoral effector inflammation.

Isolation and identification of compounds from Penthorum chinense Pursh with antioxidant and antihepatocarcinoma properties.

Penthorum chinense Pursh, widely distributed in eastern Asia, has long been used in China for both food and medicine due to its various bioactivities. The aim of this study was to isolate its active compounds with antioxidant and antihepatocarcinoma properties. P. chinense was extracted with 95% ethanol, 70% ethanol, and water, respectively, and then the 70% ethanol extract was re-extracted, resulting in petroleum ether, ethyl acetate, n-butanol, and water fractions, subsequently. Results showed that the antioxidant and antihepatocarcinoma activities of ethanol extracts were stronger than those of aqueous extract, and the ethyl acetate fraction of 70% ethanol extract showed the highest activities. Four compounds, ß-sitosterol, quercetin, pinocembrin-7-O-[3-O-galloyl-4″,6″-hexahydroxydiphenoyl]-ß-glucose (PGHG), and thonningianins A (Th A), were isolated from the ethyl acetate fraction and identified by UV, MS, and NMR spectroscopic analysis. Th A was isolated from P. chinense for the first time. PGHG and Th A exhibited higher antioxidant and antihepatocarcinoma activities than did other isolated parts of P. chinense . The antihepatocarcinoma activity of Th A was much higher than that of positive control (5-fluorouracil). PGHG and Th A were suggested to be the active chemical compositions responsible for potent antioxidant and antihepatocarcinoma properties of P. chinense , which are worthy of further study.

[In vitro effects of hemocoagulase atrix and its effective components on blood coagulation of patients with bleeding disorders].

This study was aimed to investigate the pro coagulation effects of hemocoagulase atrix and its effective components (batroxobin and factor X activator) on plasma of normal subjects and patients with bleeding disorders and their mechanisms. Activated partial thromboplastin time (APTT) and prothrombin time (PT) were measured. The factor (F)X activation and thrombin generation were analyzed by using chromogenic substrate method. The results showed that the plasma APTT of normal subjects was shortened by hemocoagulase atrix, batroxobin and FX activator, and the effect of FX activator was found to be concentration-dependent (r = 0.889, P < 0.05). The prolonged APTT of plasma from patients with bleeding disorders could be corrected by hemocoagulase atrix, batroxobin and FX activator, but PT showed no great changes resulted from the treatments. FX activator could promote FX activation and thrombin generation, while neither hemocoagulase atrix nor batroxobin showed such abilities. It is concluded that hemocoagulase atrix promotes coagulation process, and corrects coagulation abnormalities in patients with bleeding disorders, its main component batroxobin directly acts on fibrinogen, and FX activator promotes thrombin generation through activating FX.

[Clinical features and gene analyses of six patients with MYH9-related disease].

OBJECTIVE: To investigate clinical features and to identify gene mutations in six patients with nonmuscle myosin heavy chain 9 gene (MYH9)-related disease. METHODS: The platelet counts were measured using automated complete blood cell counter and manual manner. The size of platelets and inclusion bodies were observed under light microscopy. All the 40 exons and exon-intron boundaries of MYH9 gene were amplified by PCR and then DNA sequencing was performed. Restriction endonuclease analysis and polyacrylamide gel electrophoresis (PAGE) were used for polymorphism analysis. RESULTS: Six patients all shared the common features of thrombocytopenia with giant platelets and granulocyte inclusions. Four MYH9 gene mutations were found in the six patients: T97C (W33R) in exon 1, 4335Insert CAGAAGAAG (1445InsQKK) and G4269A (D1424N) in exon 30 and G5833T (E1945Stop) in exon 40. The former two were novel mutations which have not been reported in the literature. The results of restriction endonuclease analysis and PAGE could exclude the possibility of nucleotide polymorphisms. CONCLUSIONS: The MYH9 gene mutations were identified in six patients with MYH9 related disorders, and T97C (W33R) and 4335InsCAGAAGAAG (1445InsQKK) were novel mutations. MYH9 related disease should be considered in individuals with persistent thrombocytopenia which is non-responsive to corticosteroids and immuno-repressive agents.

Comparative GC/MS analysis of essential oils extracted by 3 methods from the bud of Citrus aurantium L. var. amara Engl.

UNLABELLED: A comparative study of steam distillation extraction (SDE), reflux extraction (RE), and ultrasound-assisted extraction (USE) was conducted for the extraction of essential oils from the bud of Citrus aurantium L. var. amara Engl. Each method was evaluated in terms of qualitative and quantitative composition of the isolated essential oil by gas chromatography/mass spectrometry (GC/MS). The extract yields of essential oil were 0.16%, 2.18%, and 2.34%, respectively. A total of 82 compounds were identified by GC/MS. The main components obtained by SDE were terpinen-4-ol (20.98%), dipentene (11.67%), terpinene (9.24%), those by RE were palmitic acid (20.61%), 2-chloroethyl linoleate (14.54%), tetracosane (12.26%), and α-linolenic acid (11.24%), and those by USE were tetracosane (11.32%), heneicosane (11.06%), and palmitic acid (8.76%). Comparative analysis indicated that SDE was favorable for the extraction of monoterpene hydrocarbons, sesquiterpene hydrocarbons, alcohols, and carbonyl compounds, RE and USE had certain advantages in the extraction of aliphatic saturated hydrocarbons, organic acids, and esters. It is concluded that different extraction methods may lead to different yields of essential oils; the choice of appropriate method is very important to obtain more desired components with higher physiological activities. PRACTICAL APPLICATION: C. aurantium oils from different plant parts have great economic, medicinal, and nutritional values because of their wide-spectrum biological activities. The essential oil from C. aurantium L. var amara is one of the best C. aurantium oils. The data presented in this article will help us understand the relationship between essential oils and its extraction methods and know more about the aromatic components of Citrus aurantium bud. The methods established in this study will provide useful reference information for further studies, and offer essential oil industries with helpful guidance in practice.

[Clinical and gene study of three pedigrees of phytosterolemia associated with macrothrombocytopenia and hemolysis].

OBJECTIVE: To study the clinical features and ABCG5/ABCG8 gene mutations of three pedigrees of phytosterolemia presented with macrothrombocytopenia and hemolysis. METHODS: Erythrocyte and platelet morphology were examined under light microscope. Plasma sterol levels were measured by high pressure/performance liquid chromatography method. All of ABCG5 and ABCG8 exons and intron-exon boundaries were directly sequenced to identify mutations, the corresponding gene mutation sites of three families members and healthy individuals were detected. RESULTS: All the patients presented macrothrombocytopenia, hemolysis, splenomegaly and xanthomas. The blood smears showed large platelets, some as large as erythrocytes, and abnormal erythrocyte shapes, such as stomatocytes. Plasma concentrations of phytosterols, especially sitosterol were markedly elevated (30 fold) in the affected patients. Four mutations were identified in these three pedigrees, ABCG5 C20896T (R446X) and A20883G, ABCG8 del43683-43724 and del1938C-1939G/ins1938T. The latter three were novel mutations reported for the first time. CONCLUSIONS: Phytosterolemia associated with macrothrombocytopenia and hemolysis is a new subtype of this disease. Plasma phytosterols and related gene analysis should be performed when ever an unexplained macrothrombocytopenia, especially combined with haemolysis or/and stomatocytosis.

[Analysis of clinical features and gene mutations in 6 patients with Wiskott-Aldrich syndrome].

OBJECTIVE: To investigate clinical features, laboratory alterations and gene mutations of 6 patients with Wiskott-Aldrich syndrome (WAS). METHODS: T lymphocyte subtypes were measured by flow cytometer. The routine blood tests including platelet count and mean platelet volume were performed by complete blood analyzer Sysmex XE2100. Serum immunoglobulin was measured by immunoturbidimetry. Mutations in WAS protein (WASP) gene (including all the exons and exon-intron boundaries and 3', 5' untranslation region) of 6 patients and their family members were identified by PCR and sequencing. RESULTS: The patients presented with petechiae, easy bruise, eczema, bloody diarrhea, recurrent infection and fever, and the clinical scores were 3 or 4. They were thrombocytopenia with smaller mean platelet volume, anemia and leukocytosis. Megakaryocyte number was normal or slightly increased in bone marrow. In the probands, the percentage of CD3+ T cells was decreased, the CD4+/CD8+ ratio was abnormal, while the fractions of CD19+ and CD16+ CD56+ cells were in normal range. In most of the patients, the serum levels of IgG and IgA were increased. Six mutations were identified in the patients, including 10250 C-->T, and five novel mutations: 6783 C-->G,10216-10221 Ins G, 9964 Del T,10192-10203 Del GCCTGCCGGGG and 10052-10059 del GCTACTG. The 6783 C-->G in exon 3 resulted in premature stop at Tyr102, and the remaining four mutations in exon 10 resulted in frame shift and premature stop. CONCLUSION: The main characteristics of these WAS patients were thrombocytopenia with smaller mean platelet volume and immunological disturbance. Their gene mutations were deletion, insertion or nonsense mutations. All the patients had been misdiagnosed as ITP, indicating the importance of differential diagnosis.

[Genotype and phenotype analysis of congenital coagulator factor VII deficiency in four Chinese pedigrees].

OBJECTIVE: To investigate the clinical manifestation and gene mutation in four Chinese pedigrees with the congenital coagulation factor VII deficiency. METHODS: Prothrombin time (PT), activated partial thromboplastin time, thrombin time and plasma fibrinogen were measured using STAGO STA-R automatic coagulation analyzer, and the coagulation activity of factor VII (FVII:C) was determined by a PT-based one stage method, and factor VII antigen (FVII:Ag) level by a sandwich enzyme-linked immunoabsorbsent assay. All exons, exon-intron boundaries and 3',5'untranslated regions of the FVII gene from the genomic DNA of the probands and their families were amplified by PCR, and then sequenced. RESULTS: PT was significantly prolonged, and FVII:C and FVII:Ag were decreased and the following mutations were identified in the four probands: a homozygous transversion of 18041 T→G resulting in His408→Gln substitution in exon 8 in proband 1, a homozygous double nucleotide deletion, del CT (5078 - 5079) in exon 1 in proband 2, a double heterozygous of IVS6-1G→A and Gln426→stop in proband 3, and a double heterozygous of IVS6-1G→A and Arg364Gln in prohand 4. CONCLUSION: Two missense mutations, His408Gln, Arg364Gln and one nonsense, Gln426stop in the catalytic domain of FVII and one double nucleotide deletion, del CT (5078 - 5079) in exon 1 and one splicesome mutation, IVS6-1G→A in intron 6 were separately identified in four Chinese pedigrees with inherited coagulation factor VII deficiency. The Gln426stop and IVS6-1G→A were first identified in the world and the homozygous del CT (5078 - 5079) and His408Gln were first found in China.

Immunoregulatory actions of polysaccharides from Chinese herbal medicine.

IMPORTANCE OF THE FIELD: Polysaccharides, one of main classes of bioactive substances from Chinese herbal medicine (CHM), have been indicated to have wide pharmacological activities, especially broad immunomodulatory and antitumour effects. However, their immunoregulatory mechanisms are still not fully understood yet. AREAS COVERED IN THIS REVIEW: Polysaccharides from CHM (CHMPS) are reviewed with focus on their immunoregulatory function, describing their immunoregulatory actions on immune organs, immune cells and immune molecules, and discussing their effects on cell surface receptors and cell signaling pathways. WHAT THE READER WILL GAIN: A better understanding of the immunoregulatory effects of CHMPS and their structure-function relationship. TAKE HOME MESSAGE: CHMPS can active or regulate the immune system including innate and adaptive responses, and have profound effects on different diseases. Through examining the molecular mechanisms of the immunomodulating effects of CHMPS, it can be shown that CHMPS have the potential to be an adjuvant in cancer therapies.

[Inherited dysfibrinogenemia caused by Arg16His mutation in alpha chain of fibrinogen.].

OBJECTIVE: To analyze the phenotype and genotype of a family with inherited dysfibrinogenemia. METHODS: Assays of coagulation, including activated partial thromboplastin time (APTT), prothrombin time (PT) and thrombin time (TT), were carried out with Stago Compact in the proband and his family members. The activity and antigen of fibrinogen in plasma were determined by Clauss and immunoturbidimetry, respectively. Fibrinogen and its constituent were analyzed by Western blot with nonreducing 4%-20% SDS-polyacrylamide gel electrophoresis (PAGE). All exons and exon-intron boundaries of fibringen genes FGA, FGB and FGG were analyzed by PCR and then direct sequencing. RESULTS: The proband had normal APTT and PT, but prolonged TT. The activity of fibrinogen in plasma was decreased while its antigen level was normal. These abnormalities were also found in his mother and a sister. Genetic analysis revealed heterozygous G1233A in the exon 2 of FGA originating from his mother, which resulted in Arg16His missense mutation. CONCLUSION: Inherited dysfibrinogenemia was caused by Arg16His mutation in exon 2 of FGA, and this is the first case reported in a Chinese family.