Clinical diagnosis and treatment of hereditary thrombocytopenia and purpura: a report of five cases and literature review / 中华血液学杂志

Objective: To report the clinical manifestations and laboratory features of five patients with congenital thrombotic thrombocytopenic purpura (cTTP) and explore its standardized clinical diagnosis and treatment along with a review of literature. Methods: Clinical data of patients, such as age of onset, disease manifestation, personal history, family history, and misdiagnosed disease, were collected. Treatment outcomes, therapeutic effects of plasma infusion, and organ function evaluation were observed. The relationship among the clinical manifestations, treatment outcomes, and ADAMTS13 gene mutation of patients with cTTP was analyzed. Additionally, detection of ADAMTS13 activity and analysis of ADAMTS13 gene mutation were explored. Results: The age of onset of cTTP was either in childhood or adulthood except in one case, which was at the age of 1. The primary manifestations were obvious thrombocytopenia, anemia, and different degrees of nervous system involvement. Most of the patients were initially suspected of having immune thrombocytopenia. Acute cTTP was induced by pregnancy and infection in two and one case, respectively. ADAMTS13 gene mutation was detected in all cases, and there was an inherent relationship between the mutation site, clinical manifestations, and degree of organ injury. Therapeutic or prophylactic plasma transfusion was effective for treating cTTP. Conclusions: The clinical manifestations of cTTP vary among individuals, resulting in frequent misdiagnosis that delays treatment. ADAMTS13 activity detection in plasma and ADAMTS13 gene mutation analysis are important bases to diagnose cTTP. Prophylactic plasma transfusion is vital to prevent the onset of the disease.

Analysis on Prescription Research and Historical Evolution of Mahuangtang / 中国实验方剂学杂志

Based on the ancient Chinese medical literature and modern clinical application literature， Mahuangtang was systematically analyzed， including its origin， composition， processing requirements and others， in order to provide support for the development of this famous classical formula. It was found that there were some doubts about Mahuangtang， such as the original variety， the processing of decoction pieces， and the dose conversion ratio. On the basis of fully considering the actual needs of the development of famous classical formula preparations and the usage habits of modern clinical practice， the development suggestions of the author were as follows：Ephedrae Herba should be selected Ephedra sinica without removing knots， Cinnamomi Ramulus should be selected from Cinnamomum cassia， Glycyrrhizae Radix et Rhizoma should select Glycyrrhiza uralensis and be roasted with honey， and Armeniacae Semen Amarum should be selected as Dan-processed products. The total dose of Mahuangtang was 24 g after conversion according to 1 Liang equal to 3 g as following the transition phenomenon and law. The preparation and usage method was as follows：took 1.8 L of water， first added Ephedrae Herba in water for decocting， evaporated 400 mL of water， removed the upper foam， then added the other three drugs， cooked over gentle heat to 500 mL， filtered the residue， took three times a day， about 160 mL warm decoction once. The major function recorded of Mahuangtang in ancient books was exterior sthenia syndrome of exogenous wind cold， it often used in the clinical treatment of cold， acute bronchitis， bronchial asthma and other diseases through the flexible application of the later doctors.

Acupuncture based on meridian diagnosis for chronic atrophic gastritis / 中国针灸

OBJECTIVE@#To compare the efficacy between acupoint selection of meridian diagnosis and regular acupoint selection for chronic atrophic gastritis (CAG).@*METHODS@#A total of 70 cases of CAG were randomly divided into an observation group (35 cases, 5 cases dropped off) and a control group (35 cases, 5 dropped off). In the observation group, according to the hand diagnosis of meridians and the results of 80-channels energy determinator, based on the principle of child-mother relation acupoint combination, the -connecting point and back- points were added for excess syndrome, and the -primary point, front- points and -sea point of foot meridians were added for deficiency syndrome; in addition, the acupoints of the eight extraordinary meridians were added based on the nature of acupoints. In the control group, Zhongwan (CV 12), Neiguan (PC 6), Zusanli (ST 36) and Gongsun (SP 4) were selected as the primary acupoints, and additional acupoints were added according to syndrome differentiation. The two groups were treated twice a week (Tuesday and Thursday, respectively), totally for 6 months. Six months after treatment, the follow-up was conducted. The clinical symptom score, gastroenteropathy patient reported outcomes (PRO) scale score before treatment, after treatment and during follow-up as well as the score of pathological changes of gastric mucosa before and after treatment were compared between the two groups.@*RESULTS@#After treatment and during follow-up, the clinical symptom scores and gastroenteropathy PRO scale scores were decreased in the two groups (<0.01, <0.001); at the follow-up, the gastroenteropathy PRO scale score in the observation group was lower than that in the control group (<0.01). After treatment, the scores of pathological changes of gastric mucosa in the two groups were decreased (<0.01), and the score in the observation group was lower than that in the control group (<0.05).@*CONCLUSION@#The acupoint selection of meridian diagnosis is superior to regular acupoint selection for CAG, which has better efficacy, more significant improvement on gastric mucosa pathology, and more stable long-term effect.

Analysis of the clinical effect on post-stroke shoulder hand syndrome stage Ⅰ treated with the along-meridian trochar acupuncture therapy / 针刺研究

OBJECTIVE: To compare the differences in the clinical effect on post-stroke shoulder hand syndrome (SHS) stage Ⅰ between the along-meridian trochar acupuncture therapy and the routine acupuncture therapy with filiform needles. METHODS: A total of 80 patients with post-stroke SHS stage I were divided into a treatment group (41 cases) and a control group (39 cases) according to the random number table. In the control group, the common filiform needles were used to stimulate Jianyu (LI15), Jianliao (TE14), Jianzhen (SI9), Jianzhongshu (SI15), Jianwaishu (SI14), 5 times a week, 3 weeks as 1 course. In the treatment group, along-meridian trochar acupuncture therapy was applied, 3 times a week, 3 weeks as 1 course. The patients in both groups were all treated with basic medications and routine rehabilitation training. Pain degree, edema degree, upper limb motor function and activity of daily living were observed in the two groups before the treatment, at the end of the treatment and in follow-up. At the end of treatment and in follow-up, the therapeutic effect was evaluated respectively in the patients of the two groups. RESULTS: Compared with the values before treatment, the VAS score of the upper limb was reduced obviously (P< 0.001), the score of the upper limb motor function and Barthel index were increased obviously (P<0.001, P<0.05) in the patients of the two groups, the score of edema degree of the affected limb was reduced after treatment in the patients of the treatment group (P<0.001). Compared with the control group, VAS score of the upper limb and the score of edema degree of the affected limb were obviously lower (P<0.001), and the score of the upper limb motor function and Barthel index were obviously higher in the treatment group (P<0.001). The total effective rate was 66.7% (26/39) after treatment and was 74.4% (29/39) in follow-up in the treatment group and they were 20.5% (8/39) and 28.2% (11/39) respectively in the control group. The total effective rates after treatment and in follow-up in the treatment group were all obviously higher than those in the control group respectively (P<0.001). CONCLUSION: The along-meridian trochar acupuncture therapy remarkably relieves pain and edema and improves the upper limb motor function and the activity of daily living in the patients with post-stroke shoulder hand syndrome and its clinical therapeutic effect is definite.

Clinical observation of desmopressin in the treatment of 15 patients with DDAVP / 中华血液学杂志

Objective: To assess the significance of DDAVP use in the diagnosis and treatment of VWD. Methods: An analysis of 15 VWD cases who referred to Hematology Division of First affiliated Hospital of Soochow University and treated with DDAVP from March 2016 to August 2018 was conducted. Efficacy and treatment response of DDAVP were monitored by observations of changes in factor Ⅷ procoagulant (FⅧ∶C) and von Willebrand Factor (VWF) related indicators before and 2 h after DDAVP injection. Results: Of 15 cases with VWD, 7 males and 8 females with a median age of 23 (6-46) years, 7 of 9 type I VWD patients achieved complete response (CR) , 1 type 2A VWD case CR, 5 type 3 VWD ones no response (NR) . The VWF multimer analysis in 5 patients combined with other plasma VWF values were in accordance with the known diagnosis. Conclusions: DDAVP was effective in most type 1 patients, and ineffective in some type 2 and almost all type 3 cases. It was helpful for diagnosis and subsequent treatment planning.

Preliminary study on standardization of production and processing of Scutellaria baicalensis pieces / 中国中药杂志

Decoction pieces are important raw materials in the production of traditional Chinese medicine( TCM),and their quality could directly affect the clinical efficacy and medication safety. Research on the production and processing technology of TCM is the basis for the normalization and standardization of Chinese medicine decoction pieces. At present,the production and processing standards for Scutellaria baicalensis pieces are non-regulated,lacking data foundation. In this study,with baicalin,baicalein,wogonoside and wogonin contents as evaluation indicators,single factor experiment was designed to optimize the softening,drying and cutting processes of S. baicalensis,providing a basis for the standardization of their production and processing. The effects of different softening,drying and cutting processes on the contents of the main components in S. baicalensis were comprehensively analyzed by the summation of relative differences. RESULTS:: showed that the contents of the four components and comprehensive indexes were affected by different softening methods and drying temperatures. The content of wogonin in boiling method was higher than that in boiling with cold water,and the content of glycosides in 70 ℃ drying condition was higher than that in other groups. The content of baicalin was significantly affected by different cutting thicknesses,but not by comprehensive index. Eventually,the optimal preparation process for S. baicalensis was determined as follows: boiled in boiling water for 20 min,cut into thin slices( 1-2 mm),and then dried at 70 ℃ in blast drier. This process was close to the actual production,practical and feasible and meanwhile,it was of great significance to improve the quality of S. baicalensis pieces.

Establishment of biological assess for quality control of Fuzi based on determination of premature ventricular contractions in rats / 中国中药杂志

Aconiti Lateralis Radix (Fuzi) is a toxic traditional Chinese medicine with definite efficacy. In order to improve the quality control of its different prepared products and ensure the security in clinic, it is significant to establish a method of quality evaluation related to clinic adverse effects. Aiming at the important biological marker of early cardiac toxicity reaction, there was no method to detect it. In this manuscript, a novel approach for measuring the minimal toxic dose (MTD) of premature ventricular contractions (PVC) poisoning of rats was established. Then, the determination methodology and conditions were optimized to meet the needs of the quality and biological assessment, including animal sex, weight, stability of standards and test solutions. Using this method, the MTD value of different Fuzi products were determined, such as Heishunpian, Baifupian, Zhengfupian, Baofupian, and Paotianxiong. The results showed that the MTD of Fuzi was significantly decreased after detoxification processed (P<0.05) and the MTD of Heishunpian, Zhengfupian, Baofupian and Baifupian was as much as 15.76, 22.36, 19.65 and 20.97 times to that of unprocessed Shengfuzi. In addition, Paotianxiong could not induce PVC in rats, which indicated that Paotianxiong was nontoxic and safe.This method could appropriately reflects the cardiotoxity of Fuzi and its prepared samples. Together with the chemical composition analysis, the contents of diester alkaloids were explored including aconitine, mesaconitine and hypaconitine as well as monoester alkaloids in Fuzi and its prepared products were significantly associated with PVC. Furthermore, there may be some components undetermined facilitating arrhythmia to be worth exploring. This research provides an overall and comprehensive approach to diagnose early clinical cardiotoxity and control the quality of Fuzi, which could not only be a complementary solution for the chemical evaluation, but a new method to ensure its efficacy and security of clinical application.

Effects of diammonium glycyrrhizinate on hepatic and intestinal UDP-Glucuronosyltransferases in rats: Implication in herb-drug interactions / 中国天然药物

Glycyrrhizin is a major bioactive component of liquorice, which exerts multiple biochemical and pharmacological activities and is frequently used in combination with other drugs in the clinic. Mycophenolate mofetil (MMF), an immunosuppressant widely used in transplant patients, is metabolized by UDP-glucuronyltransferases (UGTs). Although significant evidence supports that glycyrrhizin could interact with the cytochrome P450s (CYPs), few studies have addressed its effects on UGTs. The present study aimed at investigating the regulatory effects of diammonium glycyrrhizinate (GLN) on UGTs in vitro and in vivo. We found that long-term administration of GLN in rats induced overall metabolism of MMF, which might be due to the induction of UGT1A protein expression. Hepatic UGT1A activity and UGT1A mRNA and protein expression were significantly increased in GLN-treated rats. UGT1A expression levels were also increased in the intestine, contradicting with the observed decrease in intestinal UGT1A activities. This phenomenon may be attributed to different concentrations of glycyrrhetinic acid (GA) in liver and intestine and the inhibitory effects of GA on UGT1A activity. In conclusion, our study revealed that GLN had multiple effects on the expression and activities of UGT1A isoforms, providing a basis for a better understanding of interactions between GLN and other drugs.

Research on contents of anthraquinones in Cassiae Semen by principal component analysis / 中国中药杂志

Cassiae Semen is a common traditional Chinese medicine, and contents of anthraquinones of Cassiae Semen different significantly from area to area. According to Chinese Pharmacopoeia (2010 edition), only contents of aurantio obtusin and chrysophanol were used to evaluate the quality of Cassiae Semen, another data could be added later. Ten batches of Cassiae Semen from different areas were determined, and total anthraquinones, total free anthraquinones and total combined anthraquinones contents were assessed by ultraviolet visible spectrophotometer, contents of aurantio obtusin, rhein, aloe emodin, emodin, chrysophanol and physcion were determined by HPLC. After that, principal components analysis was used to evaluate these data determined previous by dimension reduction analysis. At last, the result suggests that three main components were found out, it shows that content of aloe emodin could be used to evaluate the quality of Cassiae Semen as well as contents of aurantio obtusin and chrysophanol. And Cassiae Semen from Hebei province posseses higher quality than Cassiae Semen from other different areas. All these results can provide a good reference for quality evaluating of Cassiae Semen medicinal materials at a certain extent.

The Expression of Programmed Death-1 in Circulating CD4+ and CD8+ T Cells during Hepatitis B Virus Infection Progression and Its Correlation with Clinical Baseline Characteristics

BACKGROUND/AIMS: Programmed death-1 (PD-1) expression was investigated in CD4+ and CD8+ T cells from hepatitis B virus (HBV)-infected patients at the chronic hepatitis B (CHB) infection, liver cirrhosis (LC), and hepatocellular carcinoma (HCC) stages. METHODS: PD-1 expression in circulating CD4+ and CD8+ T cells was detected by flow cytometry. The correlations between PD-1 expression and HBV viral load, alanine aminotransaminase (ALT) levels and aspartate aminotransferase (AST) levels were analyzed using GraphPad Prism 5.0. RESULTS: PD-1 expression in CD4+ and CD8+ T cells was significantly increased in both the CHB group and advanced-stage group (LC plus HCC). In the CHB group, PD-1 expression in both CD4+ and CD8+ T cells was positively correlated with the HBV viral load, ALT, and AST levels. However, in the LC plus HCC group, significant correlations between PD-1 expression and the clinical parameters were nearly absent. CONCLUSIONS: PD-1 expression in peripheral CD4+ and CD8+ T cells is dynamic, changes with HBV infection progression, and is related to HBV viral load and liver function, especially in CHB. PD-1 expression could be utilized as a potential clinical indicator to determine the extent of virus replication and liver injury.

Congenital afibrinogenemia caused by a novel insertion mutation in the FGB gene / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the genetic defect and its mechanism in a patient with congenital afibrinogenemia.</p><p><b>METHODS</b>The plasma fibrinogen activity and antigen of the patient was determined using the Clauss method and immuno-nephelometric assay, respectively. Genomic DNA was isolated from peripheral blood of the proband and his related family members. All exons and exon-intron boundaries of the three fibrinogen genes (FGA, FGB, FGG) were amplified by PCR followed by direct sequencing. Thrombin fibrin aggregation curve were detected in the plasma of the patient. Wild-type and mutation type fibrinogen vectors were constructed, and then transfected into COS-7 cells. The wild-type and mutant proteins from the culture media and cell lysates were tested by Western blot and ELISA.</p><p><b>RESULTS</b>APTT, PT, TT were significantly longer in the proband. Plasma fibrinogen activity and antigen of the patient could not be detected using the Clauss method and immuno-nephelometry, respectively. Gene analysis revealed that a novel homozygous GTTT insertion between nucleotides 2833 and 2834 in FGB exon 2 in the proband. The proband's father, mother, brother and son were heterozygous. The polymerization curves of the patient did not show a lag phase or final turbidity, compared with the normal controls. Western blot analysis showed the lack of complete half-molecules of the fibrinogen molecule and fibrinogen in patient's plasma under non-reducing conditions. It also could not detect the truncated Bβ chain under reducing conditions. Abnormal fibrinogen molecule (molecule weight>340 000) were found in transfected COS-7 cells by Western blot, which indicated that the mutation caused the abnormal intracellular fibrinogen molecule assembly. The fibrinogen band was absent in culture media transfected by the mutation. Fibrinogen levels of mutant fibrinogen were no significant different from those of wild-type fibrinogen in cell lysates by ELISA analysis [(2.47 ± 0.30) μg/ml vs (2.65±0.60) μg/ml, P=0.0889]; However, the levels of the mutant fibrinogen were statistically significant lower than those of wild type fibrinogen in culture media [(0.01 ± 0.01) μg/ml vs (3.80±0.80) μg/ml, P=0.0001].</p><p><b>CONCLUSION</b>Congenital afibrinogenemia was caused by this frameshift mutation in exon 2 of FGB. This novel mutation impaired fibrinogen assembly and secretion.</p>

Identification and functional analysis of a novel missense mutation Ser250Phe underlying congenital coagulation factor Ⅶ deficiency / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To identify and characterize a missence mutation Ser250 Phe underlying coagulation factor Ⅶ (FⅦ) deficiency in a Chinese patient and his family.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

Clinical efficacy observation on abdominal acupuncture for cyclomastopathy / 中国针灸

<p><b>OBJECTIVE</b>To verify clinical efficacy of abdominal acupuncture for cyclomastopathy.</p><p><b>METHODS</b>One hundred and twenty-one cases of cyclomastopathy were randomly divided into an abdominal acupuncture group (64 cases) and a routine treatment group (57 cases). In the abdominal acupuncture, basic treatment (including psychological counseling and Chinese patent medicine Rupixiao) and abdominal acupuncture at Zhongwan (CV 12), Xiawan (CV 10), Qihai (CV 6), Guanyuan (CV 4) and Huaroumen (ST 24) were applied at the same time. In the routine treatment group, only basic treatment was applied. Before and after the treatment, visual analogue scale (VAS) of main symptoms and WHO Quality of Life-100BREF score were observed, also clinical efficacy of both groups was compared.</p><p><b>RESULTS</b>The scores of VAS and WHO Quality of Life-100BREF in both groups had statistical significances before and after the treatment (both P < 0.05), indicating two treatments could both effectively relieve pain and improve life quality. The total effective rate was 84.4% (54/64) in the abdominal acupuncture group, which was superior to 68.4% (39/57) in the routine treatment group (P < 0.01). The abdominal acupuncture had best effect for moderate pain (P < 0.05).</p><p><b>CONCLUSION</b>The abdominal acupuncture could improve clinical symptoms and life quality of patients with cyclomastopathy.</p>

The identification and pharmacokinetic studies of metabolites of salvianolic acid B after intravenous administration in rats / 中国天然药物

AIM@#To identify and quantify the major metabolites of salvianolic acid B (SAB) after intravenous injection in rats.@*METHODS@#LC-IT/TOF-MS was used to identify the metabolites in rat bile, plasma, and urine; LC-MS/MS was used to quantify the two major metabolites.@*RESULTS@#In rat bile, plasma, and urine, nine metabolites were identified, including methylated metabolites of SAB, lithospermic acid (LSA), the decarboxylation and methylation metabolites of LSA, salvianolic acid S (SAS), and dehydrated-SAS. The t1/2 of monomethyl-SAB and LSA were both very short, and monomethyl-SAB had a larger AUC than LSA in rats.@*CONCLUSION@#Nine metabolites were found, the metabolic pathway was described, and the pharmacokinetic profiles of LSA and monomethyl-SAB were studied, thereby clarifying that methylation was the dominant metabolic pathway for SAB in rats.

Expression of platelet membrane glycoproteins in pediatric idiopathic thrombocytopenic purpura and its clinical significance / 中华实用儿科临床杂志

Objective To explore the relationship between the expression of platelet membrane glycoprotein in pediatric idiopathic thrombocytopenic purpura (ITP) and its clinical significance.Methods A modified monoclonal antibody immobilization of platelet antigen (MAIPA) method was used to detect the positive expression rates of 4 platelet membrane glycoproteins (GP Ⅰ b/Ⅸ,GP Ⅰ b,GP Ⅲ a,and GP Ⅰ b) in 80 pediatric patients with ITP.The correlation was explored between the GP positive rate and the clinical efficacy in pediatric ITP.Trying to observe the correlationship between the GP positive rate of pediatric ITP in the total,the different gender,the acute and chronic and the treatment response rate in pediatric ITP respectively.Results There was a significant difference in curative rate statistically between the GP positive group and the GP negative group(x2 =8.535,P ＜ 0.01) in 80 pediatric ITP patients,but no statistic difference in curative rate existed between the 36 female and 44 male(x2 =0.013,P ＞0.05).Markedly statistic difference was found in the female(x2 =4.433,P ＜ 0.05),the same to the male (x2 =4.156,P ＜ 0.05).Meanwhile,there was an extremely statistic difference between 67 acute and 13 chronic patients(x2 =23.513,P ＜ 0.001).Apparently statistic difference also occurred in the acute (x2 =4.157,P ＜ 0.05),but not in the chronic cases (x2 =0.410,P ＞ 0.05).Conclusions The clinical response rate is significantly correlated with the GP positive rate in pediatric ITP,but not correlated with gender.The GP positive rate can reflect the disease status of pediatric ITP to a certain extent and be used as an indicator for judging the efficacy and monitor prognosis of pediatric ITP.

Clinical features and gene analyses of six patients with MYH9-related disease / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate clinical features and to identify gene mutations in six patients with nonmuscle myosin heavy chain 9 gene (MYH9)-related disease.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>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.</p>

In vitro effects of hemocoagulase atrix and its effective components on blood coagulation of patients with bleeding disorders / 中国实验血液学杂志

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.

Prenatal diagnosis for two families of congenital factor V deficiency / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To provide genetic consulting and prenatal diagnosis for two families with congenital factor V deficiency based on the known mutations of factor V gene (G16088C and G69969T).</p><p><b>METHODS</b>Chorionic DNA was obtained at 12 weeks of gestation and analyzed to exclude maternal cell contamination through microsatellite DNA analysis. It was then amplified with PCR and sequenced to determine the presence of mutations in exons 3 and 23. Factor V activity of the blood was assayed at 22 weeks of gestation and 6 months after birth.</p><p><b>RESULTS</b>The fetus in case 1 was found to be a heterozygous carrier of the G16088C mutation, for whom factor V activity of the cord blood and peripheral blood were 15% and 53%, respectively. Fetus 2 did not carry the familiar G69969T mutation, for whom the factor V activity of cord blood and peripheral blood has measured 32% and 93%, respectively. Follow-up studies demonstrated that the two infants were both in good health without a tendency for bleeding.</p><p><b>CONCLUSION</b>In both cases, the genotypes were consistent with the phenotypes. This is the first report of prenatal diagnosis of congenital factor V deficiency.</p>

Analysis of clinical features and gene mutations in 6 patients with Wiskott-Aldrich syndrome / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate clinical features, laboratory alterations and gene mutations of 6 patients with Wiskott-Aldrich syndrome (WAS).</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

Clinical and gene study of three pedigrees of phytosterolemia associated with macrothrombocytopenia and hemolysis / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the clinical features and ABCG5/ABCG8 gene mutations of three pedigrees of phytosterolemia presented with macrothrombocytopenia and hemolysis.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>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.</p>