The detection of coagulation factor Ⅷ activity plays an important role in the diagnosis, typing, efficacy monitoring and detection of inhibitor titer in hemophilia A, acquired hemophilia A and von Willebrand disease. However, due to the diversity of detecting systems, the difference of reagent composition, the existence of interfering substances and other influence factors, the detection of coagulation factor Ⅷ activity in the laboratories in China still needs to be improved.

Objective:To investigate the inhibitory effect of esomeprazole on proliferation and chemosensitizing effect of breast cancer cells.Methods:Human MBA-MD-231, MCF-7 breast cancer cell line and human Huh7 liver cancer cell line were cultured by conventional methods; cells were treated with different concentrations of esomeprazole, and CCK8 kit was used to detect the proliferation of different tumor cells after stimulation. Cells were treated with different concentrations of esomeprazole, and the effects of esomeprazole on cell cycle of different cells were analyzed by flow cytometry. Cells were treated with different concentrations of paclitaxel and epirubicin combined with esomeprazole, and CCK8 kit was used to detect the proliferation of different tumor cells after stimulation. Measurement data were expressed as mean ± standard deviation ( ± s), and analysis of variance was used for comparison among multiple groups. Results:CCK8 results showed that esomeprazole could inhibit the proliferation of MBA-MD-231 cells, MCF-7 cells and Huh7 cells in a dose-dependent manner. Flow cytometry results showed that cells in G 0/G 1 phase were significantly increased by esomeprazole treatment. Esomeprazole can enhance the inhibitory effect of paclitaxel and epirubicin on the proliferation of MBA-MD-231 cells and MCF-7 cells, and improve the chemosensitivity. Conclusion:Esomeprazole blocks breast cancer cell MBA-MD-231, MCF-7 and liver cancer cell Huh7 in G 0/G 1 phase, thereby inhibiting cell proliferation. Esomeprazole can enhance the inhibitory effect of chemotherapeutic drugs on the proliferation of MBA-MD-231 and MCF-7 cells.

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.

Objective:To establish clinical detection of coagulation factor Ⅷ activity by chromogenic substrate assay and evaluate its clinical application.Methods:A total of 40 hemophilia Apatients, 20 acquired hemophilia A patients, 26 patients with positive lupus anticoagulant and 60 apparently healthy people were enrolled from January 2018 to May 2019 in Ruijin Hospital of Medical College, Shanghai Jiaotong University. According to Clinical and Laboratory Standards Institute(CLSI), the accuracy, within-run and between-run imprecision, lower detection limit, linear range, carryover rate, reference range, and reportable range of chromogenic substrate assay for detecting coagulation factor Ⅷ activity was evaluated, and compared with coagulation assay. The clinical application was evaluated by detecting F Ⅷ activity in acquired hemophilia A patients and patients with lupus anticoagulant by chromogenic substrate assay.Results:The results of two constant quality control products were within range provided by the manufacturer(the bias was 3.93%-6.79%). The within-run imprecision was 1.86%-2.06%(≤5%). The between-run imprecision was 4.83%-6.90%(≤15%). The chromogenic substrate assay had a good performance in sensitivity(CV=11.23%<20%). The recommended reference range was appropriate for our laboratory. The maximum dilution was 1∶16. The linear range was 5.00%-193.50% (a=1.0243, R 2=1.000). The clinical reportable range was 0.50%-387.00%. The method had a low carryover rate (0.04%). The chromogenic substrate assay had good consistency with coagulation assay in detecting coagulation factor Ⅷactivity (R 2=0.961) as well as the titer of FⅧ inhibitor(R 2=0.973).The difference of FⅧ activity in patients with lupus anticoagulant between these two assays was statistically significant(t=9.232,P<0.05). Conclusion:The chromogenic substrate assay has a good performance in clinical detection of coagulation factor Ⅷactivity with wider clinical application and less interference.

Objective:To study the impact of repeat hepatectomy for patients with post-hepatectomy recurrent hepatocellular carcinoma (HCC).Methods:The data of patients who developed post-hepatecotmy recurrent HCC and underwent repeat hepatectomy at the General Surgery Department of Beijing Tongren Hospital from May 2013 to May 2016 (the Recurrence Group), were retrospectively compared with the data from patients who underwent initial hepatectomy for HCC during the same study period (the Primary Group). The general data, perioperative data, postoperative complications and survival of the two groups were compared.Results:The primary group included 179 patients, consisting of 133 males and 46 females, aged (57.3±11.7) years, with a range from 14.0 to 84.0 years. The recurrence group included 36 patients, consisting of 30 males and 6 females, aged (55.9±11.4) years, with a range from 40.0 to 77.0 years. There were no statistically significant differences between the two groups in gender, age, hepatitis virus infection status, preoperative alpha fetoprotein, Child-Pugh score and indocyanine green retention rate at 15 min ( P>0.05). However, there were statistically significant differences ( P<0.05) between the two groups in operative time [(244.2±84.3)min vs. (283.4±66.8)min], intraoperative blood loss[(428.5±151.6)ml vs. (756.2±187.4)ml], anatomic or nonanatomic hepatectomy, single tumor or multiple tumors, and maximum tumor diameter[(5.81±2.24)cm vs. (3.69±1.55)cm]. There were no statistically significant differences between the two groups in incidences of tumor capsular invasion, tumor thrombus and degrees of tumor differentiation ( P>0.05). There were no statistically significant differences in surgical complication rates ( P>0.05), and in 1-year and 3-year overall and disease free survival rates between the two groups ( P>0.05). Conclusions:Repeat hepatectomy for recurrent HCC after hepatectomy was safe and effective. Its long-term survival outcomes were similar to first hepatectomy for HCC.

Objective:To assess the safety and feasibility of single-port robotic radical cystectomy.Methods:During May 2019 and August 2019, nine patients (8 males, 1 female) received single-port robotic radical cystectomy by the same surgeon. The average age was 65.6(56-78)years. After a 4.5-5.5 cm trans-umbilical incision was made, Lagiport was inserted. Da Vinci Si system 1 #, 2 # arms and 30° lens were applied. Radical cystectomy and bilateral pelvic lymphadenectomy were performed without additional ports. Urinary diversion was completed outside the body. Uterus and vaginal anterior walls were also resected for female patient. Results:All 9 surgeries were successfully conducted without additional ports or conversion to laparoscopic and open surgery. The average operation time was 437.8(280-600)min. Urinary diversion methods included 2 orthotopic ileal neobladder, 5 ideal conduit and 2 cutaneous ureterostomy. Average estimated blood loss was 227.8(100-450)ml, without blood transfusion. Average intestinal recovery time was 3.1(2-4)days, drainage duration was 8.3(3-16) days, and postoperative hospital stays was 7.7(6-13) days. Pathological TNM stage: T 2aN 0M 0 6 cases, T 2bN 0M 0 1 case, T 3aN 3M 0 1 case, T isN 0M 0 1 case. All surgical margins were negative. One bowel obstruction was cured with fasting and indwelling gastric tube. During 9-12 months’ follow-up, no tumor recurrence and metastasis were observed. There was no hydronephrosis or ureterostenosis. All surgical incision healed well. Conclusions:For experienced surgeons, single-port robotic radical cystectomy is safe and feasible with small incision and fast recovery. Short-term clinical result is satisfied.

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.

OBJECTIVETo perform phenotypic diagnosis, genetic diagnosis and prenatal diagnosis of inherited coagulation factor XIII (FXIII)deficiency in a Chinese family also provide a review of inherited coagulation F XIII deficiency.

METHODSThe activity levels of F XIII (F XIII:C) of proband and family members were measured by clot solubility test and REA-chrom F XIII kit. The antigen levels of F XIII(FXIII:Ag) were measured by enzyme-linked immunosorbent assay. Thrombelastography (TEG) test was used to make a comprehensive evaluation of coagulation status in the proband. All 15 exons and exon-intron boundaries of the F13A1 gene were amplified by PCR, and DNA sequencing was performed then. The mutation identified in the proband was screened in the family members. Furthermore, the related literatures were reviewed to provide a profile of clinical manifestation, gene mutations, the relationship between the mutations and phenotype, and treatments of inherited coagulation F XIII deficient cases.

RESULTSThe clot solubility test was positive in the proband. The FXIII:Ag level of the proband was less than 1% and the FXIII:C level was below the lower limit of detection (<3%). Two compound heterozygous missense mutations (p.Arg662* and p.Trp665*) were identified in the proband. Family study showed that the two mutations were both inherited from the parents. The fetus also carried two compound heterozygous mutations, the same as the proband, and was diagnosed with severe F XIII deficiency. As reported in the literatures, most mutations were missense mutations and nonsense mutations, and no hot spot was found. The clinical pattern of F XIII deficiency varied among patients, with potentially fatal consequences from severe bleeding complications.

CONCLUSIONBetter understanding of F XIII biochemical properties and function and developing of FXIII laboratory assays and genetic detection could prevent missed diagnosis, and patients moght benefit from better care.

Asian Continental Ancestry Group ; Base Sequence ; DNA Mutational Analysis ; Enzyme-Linked Immunosorbent Assay ; Exons ; Factor XIII ; genetics ; Factor XIII Deficiency ; genetics ; Female ; Heterozygote ; Humans ; Introns ; Mutation, Missense ; Pedigree ; Phenotype ; Polymerase Chain Reaction ; Pregnancy ; Prenatal Diagnosis

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.

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.