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Objective: To analyze the phenotype and genotype of two pedigrees with inherited fibrinogen (Fg) deficiency caused by two heterozygous mutations. We also preliminarily probed the molecular pathogenesis. Methods: The prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT) and plasma fibrinogen activity (Fg∶C) of all family members (nine people across three generations and three people across two generations) were measured by the clotting method. Fibrinogen antigen (Fg:Ag) was measured by immunoturbidimetry. Direct DNA sequencing was performed to analyze all exons, flanking sequences, and mutated sites of FGA, FGB, and FGG for all members. Thrombin-catalyzed fibrinogen polymerization was performed. ClustalX 2.1 software was used to analyze the conservatism of the mutated sites. MutationTaster, PolyPhen-2, PROVEAN, SIFT, and LRT online bioinformatics software were applied to predict pathogenicity. Swiss PDB Viewer 4.0.1 was used to analyze the changes in protein spatial structure and molecular forces before and after mutation. Results: The Fg∶C of two probands decreased (1.28 g/L and 0.98 g/L, respectively). The Fg∶Ag of proband 1 was in the normal range of 2.20 g/L, while it was decreased to 1.01 g/L in proband 2. Through genetic analysis, we identified a heterozygous missense mutation (c.293C>A; p.BβAla98Asp) in exon 2 of proband 1 and a heterozygous nonsense mutation (c.1418C>G; p.BβSer473*) in exon 8 of proband 2. The conservatism analysis revealed that Ala98 and Ser473 presented different conservative states among homologous species. Online bioinformatics software predicted that p.BβAla98Asp and p.BβSer473* were pathogenic. Protein models demonstrated that the p.BβAla98Asp mutation influenced hydrogen bonds between amino acids, and the p.BβSer473* mutation resulted in protein truncation. Conclusion: The dysfibrinogenemia of proband 1 and the hypofibrinogenemia of proband 2 appeared to be related to the p.BβAla98Asp heterozygous missense mutation and the p.BβSer473* heterozygous nonsense mutation, respectively. This is the first ever report of these mutations.

In March 2017, a severely burned male patient aged 36 years with hypovolemic shock was admitted to our hospital. The patient received large quantities of antibiotics and blood products and repeated skin graft after admission, and then he suffered wound errhysis and throat congestion. The patient was healthy before without family history of bleeding or thrombosis disease. Laboratory tests showed that prothrombin time and activated partial coagulation time were remarkably prolonged, blood coagulation factor Ⅴ activity was extremely low, and the result of qualitative test of coagulation factor inhibitor was positive. Acquired blood coagulation factor Ⅴ deficiency was diagnosed. After application of dexamethasone (5 mg, twice per day) and infusion of fresh frozen plasma, blood coagulation indicators of patients recovered in 4 days, the result of qualitative test of coagulation factor inhibitor was negative, and bleeding symptoms were improved.

Introduction: The dormancy of Davidia involucrata seeds normally lasts for an extended period of time and because of this unique property the species is an excellent model for studying the molecular mechanisms of plant dormancy. The use of minimal plant material is desirable for RNA extraction since D. involucrata is a rare plant and it is relatively difficult to collect large amounts of seeds in order to perform molecular biology studies., Objective: To improve the quality of RNA obtained from seeds of D. involucrata by eliminating the oxidation of polyphenols during extraction and by preventing polysaccharides and other impurities from being extracted., Methods: A previously described method was modified by the addition of 4% (w/v) poly(N-vinyl-2-pyrrolidone) to the dry seeds during grinding and by adding 5% (v/v) beta-mercaptoethanol and 28% (v/v) ethanol to the extraction buffer. Two further centrifugation steps (5000 and 8000 rpm) were utilised and one-seventh volume of ethanol was incubated with the supernatant at 4 degrees C for 2-3 h prior to the precipitation of RNA., Results: Following these modifications, an effective method was established for total RNA extraction from a small amount of dry seeds of D. involucrata. The isolated RNA was shown to have high purity and integrity by gel electrophoresis and spectrophotometry, and was confirmed to be suitable for RT-PCR and the construction of cDNA libraries., Conclusion: The modified method reduced the amount of seeds required for extraction of total DNA and was beneficial for preserving the endangered species., (Copyright (c) 2009 John Wiley & Sons, Ltd.)

A amino-terminated G 4.0 dendrimer-like polyamidoamine (PAMAM) immobilized silica gel (PAMAMSG) was prepared with a divergent method by repeating two processes: (1) Michael addition of methyl acrylate (MA) to surface amino groups; and (2) amidation of the resulting esters with ethylenediamine (EDA) from gamma-aiminopropyl silica gel (APSG) core. It was then used for the first time as microcolumn packing for the flame atomic absorption spectrometry (FAAS) determination of trace or ultra trace Pd(II), after flow injection (FI) online preconcentration and separation process. A limit of detection (LOD) of 3.9ngml(-1) was achieved when 0.200mugml(-1) Pd(II) was preconcentrated in 0.2moll(-1) HCl medium with a sampling flow rate of 6.0mlmin(-1) for 60s and the relative standard deviation (R.S.D.) was 1.7%. The proposed method was successfully used for the determination of Pd in two metallurgical samples.

Background: The high incidence of residual stones has been a major problem in the treatment of hepatolithiasis. Although various imaging techniques have been used to locate the stones, and many different postoperative procedures have been used as remedial modalities to remove the residual calculi, results have been far from satisfactory., Objectives: To remove obstinate hepatic stones and reduce the incidence of residual calculi., Design: Prospective clinical trial., Setting: Medical university-affiliated hospital., Patients and Methods: Ten patients who had residual intrahepatic stones after conventional operative procedures underwent intraoperative ultrasound (IOUS)-guided transhepatic lithotomy between July 1988 and July 1995. This surgical technique includes accurately locating stones with IOUS, choosing a surgical approach path under the guidance of IOUS while avoiding critical blood vessels and uninvolved biliary tracts, dividing hepatic parenchyma to the involved biliary ducts, and removing the obstinate calculi using the real-time image of IOUS, which is able to monitor the movement of the lithotomy instruments without interruption., Main Outcome Measures: Clinical practical value of IOUS-guided transhepatic lithotomy in the treatment of residual hepatic stones., Results: Complete removal of the stones was achieved in all 10 patients. There were no severe complications or mortality at a median follow-up of 39 months., Conclusions: IOUS-guided transhepatic lithotomy can greatly decrease the incidence of residual hepatic stones. It is accurate and safe. As a new surgical procedure, IOUS-guided transhepatic lithotomy should be an alternative modality in the management of hepatolithiasis, although the long-term benefits still need to be observed.

Complete clearance of intrahepatic stones has long been a major surgical challenge. To reduce the incidence of residual stones, we used intraoperative ultrasonography (IOUS) to localize them and guide lithectomy in 38 patients with hepatolithiasis between July 1988 and December 1993. All patients had multiple intrahepatic stones; 24 had accompanying extrahepatic calculi. Hepatic stones were confined to the left intrahepatic biliary tract in 13 patients, to the right in 9, and in both lobes in 16. Twenty-three patients underwent common bile duct exploration followed by T-tube drainage, 8 had transhepatic lithotomy with or without choledocholithotomy, 3 had choledocolithotomy and Roux-en-Y side-to-side choledochojejunostomy, and 2 had hepaticojejunostomy, left lobectomy was performed in the remainder. In 35 patients cholecystectomy was performed at the same time. Complete clearance of the stones was achieved in 36 patients (94.7%). The incidence of retained stone was decreased to 5.3%. No associated complications occurred. IOUS can accurately localize intrahepatic calculi, directly orient lithotomy instruments to approach the stones, demonstrate the spatial relation between stone and intrahepatic critical structure, and thereby choose an optical route for transhepatic lithotomy. Imaging can be repeated at any time with no radiation exposure to the patient or the medical staff.

Based on the principles of Shafik's prostatic commissurotomy, we have developed a new transurethral prostatic commissurotomy, which is characterized by cutting, layer by layer and section by section, the urethral mucosa, the bladder muscle layer and the prostate tissue within combined part of the urethra and prostate and by cutting the prostatic capsule with electrocoagulation. The comparison of clinical objective indexes has proved that the method is applicable to high risk patients with prostate hyperplasia and neck contracture leading to urethral obstruction after TURP.

On the basis of anatomical studies of renal vessels, segmental polar nephrectomy has been designed. The key point is to identify blood-loss area of the segmental pole by interruption of the upper or the lower segmental polar artery before partial nephrectomy. The merits of the procedure are non-compression of the renal pedicle vessels, appropriate extent of the segmental polar nephrectomy and less hemorrhage.