1.Extract Astragalus Polysaccharides by Ultrasonic Cell Disrupter
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(03):-
Objective To study the effect on the extract of Astragalus polysaccharides by ultrasonic cell disrupter. Methods Astragalus polysaccharides was extracted by Ultrasonic Disrupter,boiling in water and soakage in alkali,and compared the content of polysaccharides respectively. Result The content was the highest by ultrasonic disrupter,the second was by boiling in water,the worst was by soakage in alkali. Conclusion Ultrasonic disrupter is suitable to extract Astragalus polysaccharides.
2.Study on hemostatic and coagulative variations during orthotopic liver transplantation
Xuefeng WANG ; Jian HUANG ; Guanqun XU
Chinese Journal of Organ Transplantation 2003;0(06):-
Objective To investigate the hemostatic and coagulative variations during orthotopic liver transplantation (OLT).Method The blood platelet count, coagulant and anti-coagulant functions were assessed pre- and intra-operation of OLT.Results During the operation, activated partial thromboplastin time (aPTT) and prothrombin time (PT) were prolonged, platelet count (PLT), activities of most of the coagulation factors and levels of antithrombin (AT), plasminogen (PLG), plasminogen activator inhibitor-1 (PAI-1) and ?2 -antiplasmin (?2-AP) were reduced, while the levels of tissue plasminogen activator (t-PA), plasmin-?2-antiplasmin complex (PAP) and thrombin-antithrombin complex (TAT) were increased. The variations in the neohepatic phase were more significant than that in the pre-operation phase. Conclusion In the entire process of OLT operation, the coagulant and anti-coagulant functions were decreased, and the fibrinolytic functions were sthenic in the anhepatic phase and the neohepatic phase.
3.Establishment of variables-specific reference ranges of von Willebrand factor antigen and activity
Lili LI ; Wei ZOU ; Guanqun XU ; Liwei ZHANG ; Yun SHEN ; Linmin XU ; Xiaohong CAI ; Xuefeng WANG
Chinese Journal of Blood Transfusion 2017;30(7):690-692
Objective To analyze the screening results of yon Willebrand factor among patients before blood transfusion in Ruijin Hospital and discuss von Willebrand factor in ABO blood group and the relationship between age and gender,refine the classification of vWF antigen and activity by reference factors.Methods The von Willebrand factor among 247 cases of patients before blood transfusion in Ruijin Hospital with no clinical manifestations of abnormal blood clots and routine coagulation as laboratory tests for normal surgical patients.The vWF:Ag and vWF:Act were measured by immune turbidimetric method and ABO blood group was identified by blood type serology.Furthermore,the differences between A,B,O,AB different blood groups,sex and high (≥40 years) and low age group (<40 years) were compared by statistical methods.Results The levels of vWF:Ag in different blood groups were as follows:A blood type:98.5-142.00,B blood type:97.90-160.30,O blood type:82.13-125.45,A B blood type:103.00-135.80.The levels of vWF:Act in different blood groups were as follows:A blood type:76-130.14,B blood type:78.06-144.3,O blood type:60.89-116.11,AB blood type:88.99-124.09.O blood type vWF:Ag and vWF:Act were lower significantly (P<0.05) than non-O blood type,the difference was.Besides,young vWF:Ag and vWF:Act were lower significantly than in the elderly.There was no significant difference in vWF:Ag and vWF:Act levels between male and female groups.At last,the reference range of four groups of vWF activity (antigen) was obtained.Conclusion Plasma vWF antigen and activity levels were significantly affected by ABO blood type and age,and the refined reference range established for these influencing factors was beneficial for more detailed diagnosis of VWD and predicting vWF levels associated with bleeding and thrombosis risk.
4.Genetic diagnosis of four combined deficiency of factor Ⅴ and Ⅷ patients
Yeling LU ; Xuefeng WANG ; Qiulan DING ; Jing DAI ; Guanqun XU ; Dandan HUANG ; Xiaodong XI ; Hongli WANG
Chinese Journal of Laboratory Medicine 2010;33(7):618-624
Objective To investigate the genetic diagnosis and molecular pathogenesis of four patients with combined deficiency of coagulation factor Ⅴ and Ⅷ and their family members. Methods The APPT, FT, FⅤ: C, FⅧ: C were detected for phenotypic diagnosis. Thrombin generation assay was applied to determine the generation condition of thrombin in patients and healthy controls. Cenomic DNA was extracted from peripheral blood using the TianGen RelaxCene Blood DNA System;amniotic fluid DNA was extracted with phenol-ethyl ether method. The LMAN1 and MCFD2 genes were analyzed by PCR. Gene mutations were detected with nucleotid sequences by using end-labeling dideoxy method. Results The APTT of Proband 1 was significantly prolonged to 88. 2s and her PT was prolonged to 19. 6 s. The combined deficiency was identified with FⅧ (FⅧ: C 24. 2% ) and FV(FⅤ: C 9. 1% ). Proband 2 and 3 were sisters. The coagulation studies revealed that both of them had prolonged APTT (71.6 s and 74.6 s respectively) and PT (22. 1 s and 18. 3 s respectively). The combined deficiency of FⅤ (FⅤ: C 7. 6% and 14. 5% respectively) and FⅧ( FⅧ: C 25% and 19.6% respectively) were identified. Proband 4 was detected to have the prolonged APTT (70.3 s),PT (18.2 s) and the deficiency of FⅤ(FⅤ: C 9. 4% ) and FⅧ (15. 7% ). The remaining phenotype indicators test of the 4 probands were normal. The diagnosis for the 4 probands was combined deficiency of factor Ⅴ and Ⅷ. The proband 1 was detected to have compound heterozygous mutations in LMAN1 gene while having the LMAN1 and MCFD2 direct gene sequencing. One mutation was a small insertion located on exon 8 [ nt912insA (X71661. 1)] that resulted in p. 305frameshiftX20 and her mother was detected to have the same heterozygous mutation on the the locus. The other mutation was located on exon 11: nt1366C > CT ( X71661. 1 ) , p. 456Arg > Stop which was inherited from her father. Amniocyte DNA was detected to have only one heterozygous mutaion [nt1366C > CT (X71661. 1) , 456Arg > Stop] inherited from the father. No mutation in MCFD2 gene was found in proband 1 and her parents. The analysis of the MCFD2 gene in proband 2 and 3 revealed a novel homozygous single base substitution (nt411T>C) in exon 4, which results in the exchange of the amino acid isoleucine by the amino acid threonine at amino acid position 136 (p. Ile136Thr). Sequencing of the whole LMAN1 gene showed that the proband 4 had one homozygous nonsence mutation in the exon 5 of the LMAN1 ( nt615C >T,p. 202 Arg> Stop). All of the 4 probands with combined deficiency of FⅤ and FⅧ showed declined endogenous thrombin potential in the thrombin generation tests. Conclusion The combined deficiency of FⅤ and FⅧ in the proband 1 results from the compound heterozygous mutations ( nt1366C > CT and nt912insA) in LMAN1 gene, which are inherited from her parents respectively. The prenatal genetic investigation for the patient mother with preganency indicates that the fetus is a female carrier with one mutation (nt1366C > CT) inherited from the father. The homozygous missence mutation ( nt411T > C, p. Ile136Thr) in the MCFD2 gene accounts for the proband 2 and 3. The daughter of the proband 2 is a carrier with a heterozygous mutation inherited from her mother. The homozygous nonsence mutation in the LMAN1 gene of the proband 4 results in the deficency of F Ⅴ and FⅧ.
5.Molecular analysis of the genotypes and phenotypes in three pedigrees with inherited antithrombin defidency
Yingting WU ; Guanqun XU ; Liwei ZHANG ; Jing DAI ; Qiulan DING ; Xiaodong XI ; Xuefeng WANG ; Hongli WANG
Chinese Journal of Laboratory Medicine 2009;32(11):1257-1261
Objective To investigate the clinical phenotype and genotype in three probands with antithmmbin(AT)deficiency and their families,and to identify the molecular mechanism of AT deficiency.Methods Chromogenic substrate method and immunoturbidimetry assay was used to detect the plasma levels of AT:A and AT:Ag,respectively.Genomic DNA was extracted from the peripheral blood.All 7 exons and the flanking sequences were amplified by PCR.and the abnormal mutant genes were analyzed by direct sequencing.Western blot was used to detect the AT levels and thrombin generation tests were used to detect coagulation status.Results The plasma levels of AT:A and AT:Ag of the three probands declined by 50%.G7386C(Trp225Cys)mutation in exon 4,C2591G(Ser36stop)in exon 2 and C9819T(Arg359stop)in exon 5 were characterized in the three prebands and they could result in W(Trp)225C(Cys)missense mutation,S(Set)36X(stop)nonsense mutation and R(Arg)359X(stop)nonsense mutation respectively,The testing results of phenotype and genotype from some of their family members showed consistent with results from the probands.Western blot results indicated that the Icyels of PC:Ag were lower compared with the normal pooled plasma.The hypercoagulative status was present in the probands using thrombin generation tests.Conclusions Type Ⅰ hereditary AT deficiency was found in these three families.The 3 heterozygous mutations.W225C,S36X and R359X are genetic defects of hereditary AT deficiency.W225C and S36X have not been described before.
6.Mutation analysis of coagulation factor Ⅺ gene of inherited factor Ⅺ deficiency
Leiming DONG ; Qiulan DING ; Wenman WU ; Xuefeng WANG ; Guanqun XU ; Hongli WANG
Chinese Journal of Laboratory Medicine 2009;32(8):915-919
Ⅺ deficiency in Chinese Han population. Conclusion The 13 mutations of the F Ⅺ gene which were found in this study may unravel the molecular pathogenesis of the F Ⅺ deficiency in Chinese Han population.
7.Perioperative treatment effects of colorectal cancer in elderly patients
Fanghong LI ; Zhixia LI ; Dali AN ; Xiao CHEN ; Xiaohu ZHANG ; Guanqun LI ; Xu NIU
Chinese Journal of Geriatrics 2010;29(3):226-228
Objective To explore the clinical features and perioperative treatment effects of colorectal cancer in elderly patients.Methods Retrospective analysis of clinical data especially focusing on perioperative treatment from 104 elderly patients with coloreetal cancer were performed.Results A total of 99 patients (95.2%) were cured or improved at discharge.Perioperative pulmonary infection occurred in 13 cases (12.5%), of whom 6 cases (5.8%) suffered from respiratory failure.Lower extremity deep venous thrombosis occurred in 3 cases (2.9%), incision infection in 6 cases (5.8%), wound dehiscence in 2 cases (1.9%), intestinal fistula in 1 case (1.0%) and gastroparesis in 1 case (1.0%).About 5 cases (4.8%) died from multiple organ failure.Conclusions Old age is not a contraindication in surgical treatment for colorectal cancer.Paying attention to accurate and timely treatment in perioperative peroid can reduce postoperative complications and improve patients' quality of life.
8.The phenotypic and genotypic diagnosis of three Chinese patients with von Willebrand disease
Linlin JIANG ; Yanan CAO ; Xuefeng WANG ; Qiulan DING ; Guanqun XU ; Liwei ZHANG ; Jing DAI ; Yeling LU ; Hongli WANG ; Xiaodong XI
Chinese Journal of Internal Medicine 2012;51(10):788-792
Objective To analyze the phenotype and genotype of three patients with yon Willebrand disease (vWD),and to explore its molecular pathogenesis.Methods Bleeding time (BT),APTT,ristocetin induced platelet aggregation (RIPA),von Willebrand factor (vWF):ristocetin cofactor (Rco)(vWF∶ Rco),vWF antigen (vWF∶ Ag),vWF activity (vWF∶ A) test,vWF collagen binding assay (vWF∶ CB) and multimer analysis were detected for phenotype diagnosis.The dynamic process of blood coagulation was evaluated by using the thrombelastography.Genomic DNA was extracted from the peripheral blood.The vWF gene mutation was detected by sequencing.Results APTT,BT were prolonged in the three probands.Plasma vWF∶ Rco,vWF∶ Ag,vWF∶ A and vWF∶ CB were decreased in different degrees.RIPA was reduced in probands B and C.vWF multimer analysis found the lost of the large molecular weight multimers in proband B,while basically normal in probands A and C.The dynamic process of blood coagulation of proband C presented obvious hypocoagulability by using the thrombelastography.Heterozygous missense mutation g.106782G > T resulting in Cys1130Phe in exon 26,g.110988G > A resulting in Gly1579Arg in exon 28 and g.110373C >T resulting in Arg1374Cys in exon 28 were found in the probands A,B and C,respectively.Conclusion Three probands were diagnosed as type 1,type 2A or type 2MvWD by phenotype detection.Heterozygous missense mutation Cys1130Phe,Gly1579Arg and Arg1374Cys induced vWD of three probands,respectively.
9.Compound heterozygous mutations of a family with inherited hypofibrinogenemia
Linlin JIANG ; Xuefeng WANG ; Qiulan DING ; Qi OUYANG ; Guanqun XU ; Liwei ZHANG ; Jing DAI ; Yeling LU ; Xiaodong XI ; Hongli WANG
Chinese Journal of Laboratory Medicine 2012;35(4):322-327
Objective To analyze the phenotype and genotype of a Chinese family with inherited hypofibrinogenemia,and to investigate its molecular mechanism.Methods Peripheral blood was collected from seven people of this family and then plasma was separated.Activated partial thromboplastin time ( APTT),prothrombin time ( PT),thrombin time ( TT),reptilase time ( RT),the activities of antithrombin( AT∶ A ),protein C ( PC ∶ A ) and protein S ( PS ∶ A ) were tested.The activity and antigen of plasma fibrinogen were analyzed by Clauss method and immunoturbidimetry method,respectively.The fibrinogen peptide chain of the proband was semiquantitatively assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE).Thrombin generation test was performed by calibrated automated thromhogram.The dynamic process of blood coagulation was evaluated by the thrombelastography (TEG).Genomic DNA was extracted from the peripheral blood.The sequences of all the exons and exon-intron boundaries of the three fibrinogen genes FGA,FGB and FGG were amplified by polymerase chain reaction ( PCR ) and analyzed by direct sequen(c)ing.Results The activity and the antigen levels of the proband' s plasma fibrinogen were reduced to 0.48 g/L and 0.68 g/L,respectively.TT prolonged to 29.2 s and RT prolonged to 75.8 s.The assays of SDS-PAGE showed no abnormal molecular weight of fibrinogen.Peak height of thrombin generation was reduced to 249.93 nmol/L and endogenous thrombin potential was reduced to 1007.0 nmol · L-1 · min.Hypocoagulability state of the whole blood was found by TEG test.The coagulation index was - 8.6.The proband was diagnosed as inherited hypofibrinogenemia by phenotype analysis.Two mutations (Gln143Pro and g.4642delC) were found in the proband's fibrinogen Aa-chain gene,Gln143Pro came from her mother and g.4642delC came form her father.Conclusion Compound Heterozygous Mutations (Gln143Pro and g.4642delC ) of fibrinogen Aa-chain causes the proband congenital hypofibrinogenemia.
10.The preliminary application of extraperitoneal robot-assisted single-port laparoscopic radical prostatectomy
Guanqun JU ; Zhijun WANG ; Jiazi SHI ; Zhiping CAI ; Zongqin ZHANG ; Zhenjie WU ; Bing LIU ; Linhui WANG ; Dongliang XU
Chinese Journal of Urology 2021;42(1):61-62
There are few reports on the study of extraperitoneal robotic single-port laparoscopic radical prostatectomy in China. In this study, patients with localized prostate cancer were treated with extraperitoneal robotic single-port laparoscopic radical prostatectomy extraperitoneal robot-assisted single-port laparoscopic radical prostatectomy(EpRA-spRP)from April 2019 to June 2019.All patients performed EpRA-spRP successfully without adding additional auxiliary port. The operation time and blood loss were controllable, and hospitalization time was short. It is safe and feasible to perform EpRA-spRP for medium and low-risk prostate cancer. The short-term tumor control and functional recovery are satisfactory.However, the long-term effect needs further follow-up and observation.