Objective:To explore the clinical characteristics and genetic variants underlying Hereditary coagulation factor Ⅴ (FⅤ) deficiency in two Chinese pedigrees.Methods:Seventeen individuals from three generations of the two pedigrees affected with FⅤ deficiency whom had visited Taizhou Hospital of Zhejiang Province respectively in March and June 2024 were recruited as study subjects. One hundred healthy individuals undergoing physical examinations have served as the controls. Relevant coagulation parameters were measured. Thrombin generation was assessed using the calibrated automated thrombogram (CAT) assay. All exons and flanking regions of the F5 gene were amplified by PCR and directly sequenced. Candidate variants were analyzed for evolutionary conservation and potential pathogenicity, and their effects on protein structure were predicted. This study was approved by the Medical Ethics Committee of Taizhou Hospital of Zhejiang Province (Ethics No.: 20230722). Results:The FⅤ activity (FⅤ: C) and antigen levels (FⅤ: Ag) of both probands showed concurrent decrease. By thrombin generation assay, both the lag time ratio and time to peak ratio were significantly increased. Genetic analysis revealed that proband A carried compound heterozygous missense variants c. 911G>A (p.Gly304Glu) and c. 1238T>C (p.Met413Thr), whilst Proband B carried compound heterozygous missense variants c. 1258G>T (p.Gly420Cys) and c. 1538G>A (p.Arg513Lys) of the F5 gene. Conservation analysis revealed that the amino acid residues p. Gly304, p. Gly420, and p. Arg513 are highly conserved across various species. Online bioinformatics tools predicted that both the p. Gly304Glu and p. Gly420Cys variants are pathogenic. Protein modeling demonstrated that all four variants can result in alterations of protein structure or disruption of hydrogen bonding. Conclusion:FⅤ deficiency in these two pedigrees can be attributed to the compound heterozygous variants p. Gly304Glu/p.Met413Thr and p. Gly420Cys/p.Arg513Lys of the F5 gene.

OBJECTIVE: To explore the clinical characteristics and genetic variants underlying Hereditary coagulation factor V (FV) deficiency in two Chinese pedigrees. METHODS: Seventeen individuals from three generations of the two pedigrees affected with FV deficiency whom had visited Taizhou Hospital of Zhejiang Province respectively in March and June 2024 were recruited as study subjects. One hundred healthy individuals undergoing physical examinations have served as the controls. Relevant coagulation parameters were measured. Thrombin generation was assessed using the calibrated automated thrombogram (CAT) assay. All exons and flanking regions of the F5 gene were amplified by PCR and directly sequenced. Candidate variants were analyzed for evolutionary conservation and potential pathogenicity, and their effects on protein structure were predicted. This study was approved by the Medical Ethics Committee of Taizhou Hospital of Zhejiang Province (Ethics No.: 20230722). RESULTS: The FV activity (FV: C) and antigen levels (FV: Ag) of both probands showed concurrent decrease. By thrombin generation assay, both the lag time ratio and time to peak ratio were significantly increased. Genetic analysis revealed that proband A carried compound heterozygous missense variants c.911G>A (p.Gly304Glu) and c.1238T>C (p.Met413Thr), whilst Proband B carried compound heterozygous missense variants c.1258G>T (p.Gly420Cys) and c.1538G>A (p.Arg513Lys) of the F5 gene. Conservation analysis revealed that the amino acid residues p.Gly304, p.Gly420, and p.Arg513 are highly conserved across various species. Online bioinformatics tools predicted that both the p.Gly304Glu and p.Gly420Cys variants are pathogenic. Protein modeling demonstrated that all four variants can result in alterations of protein structure or disruption of hydrogen bonding. CONCLUSION FV deficiency in these two pedigrees can be attributed to the compound heterozygous variants p.Gly304Glu/p.Met413Thr and p.Gly420Cys/p.Arg513Lys of the F5 gene.
Adult ; Female ; Humans ; Male ; Middle Aged ; China/ethnology* ; Factor V/chemistry* ; Factor V Deficiency/genetics* ; Heterozygote ; Pedigree ; East Asian People/genetics*

Objective:To explore the clinical characteristics and genetic variants underlying Hereditary coagulation factor Ⅴ (FⅤ) deficiency in two Chinese pedigrees.Methods:Seventeen individuals from three generations of the two pedigrees affected with FⅤ deficiency whom had visited Taizhou Hospital of Zhejiang Province respectively in March and June 2024 were recruited as study subjects. One hundred healthy individuals undergoing physical examinations have served as the controls. Relevant coagulation parameters were measured. Thrombin generation was assessed using the calibrated automated thrombogram (CAT) assay. All exons and flanking regions of the F5 gene were amplified by PCR and directly sequenced. Candidate variants were analyzed for evolutionary conservation and potential pathogenicity, and their effects on protein structure were predicted. This study was approved by the Medical Ethics Committee of Taizhou Hospital of Zhejiang Province (Ethics No.: 20230722). Results:The FⅤ activity (FⅤ: C) and antigen levels (FⅤ: Ag) of both probands showed concurrent decrease. By thrombin generation assay, both the lag time ratio and time to peak ratio were significantly increased. Genetic analysis revealed that proband A carried compound heterozygous missense variants c. 911G>A (p.Gly304Glu) and c. 1238T>C (p.Met413Thr), whilst Proband B carried compound heterozygous missense variants c. 1258G>T (p.Gly420Cys) and c. 1538G>A (p.Arg513Lys) of the F5 gene. Conservation analysis revealed that the amino acid residues p. Gly304, p. Gly420, and p. Arg513 are highly conserved across various species. Online bioinformatics tools predicted that both the p. Gly304Glu and p. Gly420Cys variants are pathogenic. Protein modeling demonstrated that all four variants can result in alterations of protein structure or disruption of hydrogen bonding. Conclusion:FⅤ deficiency in these two pedigrees can be attributed to the compound heterozygous variants p. Gly304Glu/p.Met413Thr and p. Gly420Cys/p.Arg513Lys of the F5 gene.

Corona virus disease 2019 (COVID-19) patients with trauma are at high risk of venous thromboembolism (VTE), which must be taken seriously in the therapeutic processes. Hypercoagulable state is induced by 2019 novel coronavirus (2019-nCoV) in many ways, such as increasing the level of inflammatory factors and fibrinogen, and inducing endothelial cell injury. The venous wall injuries from trauma and operation directly or indirectly trigger off the exogenous coagulation pathway and the microcirculation can be damaged at the same time, which may initiate the exogenous pathway of VTE. Immobilization of limbs and forced bed rest during the treatment of traumatic patients will slow venous blood flow. Chronic non-communicable diseases such as diabetes in the elderly were independent risk factors for VTE. Furthermore, the persistent fever, severe lung disease, respiratory failure, sepsis and invasive technology application add the risk of VTE and the difficulty of treatment. In order to help effective prevention VTE of for COVID-19 patients with trauma, the authors put forward relevant technical suggestions for prevention and nursing of VTE to provide basis for nursing work during pandemic of COVID-19.

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.

Objective: To determine the prevalence and antimicrobial susceptibility of Salmonella isolated from broiler production process in 4 provinces of China. Methods: Using convenience sampling method, 238 sample sites from broiler whole production process were chosen in Henan, Jiangsu, Heilongjiang and Shandong provinces in 2012. A total of 11 592 samples were collected and detected to analyze prevalence baseline, including 2 090 samples from breeding chicken farms and hatcheries, 1 421 samples from broiler farms, 5 610 samples from slaughterhouses and 2 471 samples from distribution and retail stores. All Salmonella strains were isolated through selective enrichment, and were serotyped according to Kauffmann-White scheme. The antimicrobial susceptibilities of selected Salmonella strains were determined by the broth microdilution method and fourteen antimicrobial agents were examined. Results: During incubation course, the average prevalence of Salmonella was 5.5% in feces of breeding hens, feces of chicks, and hatching eggs, 123 Salmonella strains were isolated. During cultivation course, the prevalence of Salmonella was 8.0% in feces from broiler farms, soil, feed, and workers, 114 Salmonella strains were isolated. During slaughter course, the prevalence of Salmonella was 24.9% in swabs pre-slaughter, dressed broiler carcasses, pre-cooled broiler carcasses, water from precooling pool, cutter and chipping boards, frozen chicken portions, and workers, 1 438 Salmonella strains were isolated. During distribution and sale course, the prevalence of Salmonella was 20.9% in transport carts, frozen chicken portions, retail chicken portions and workers, 551 Salmonella strains were isolated. The dominant Salmonella serotypes were Salmonella Enteritidis (n=1 229) and Salmonella Indiana (n=621). Among 1 231 examined strains, 97.2% Salmonella isolates were resistant to at least one antimicrobial, 69.9% Salmonella strains were multi-drug resistant isolates. Conclusion Our findings indicated that Salmonella contamination was common and serious in commercial broiler whole production process in China, especially in the course of defeathering, precooling and selling. The environment of broiler farm is the important source of Salmonella contamination. Additionally, antibiotic resistance of Salmonella was serious for common antimicrobials and multi-drug resistant strains existed widespread, which can pose potential risk on public health and clinical therapy.

Objective To investigate the effect of autologous tissue breast reconstruction with microsurgical lymph node transfers and lymphatic-venous anastomoses for the treatment of mastectomy related axillary cavitydeformation and upper extremity lymphedema .Methods The donor sites of lymph node transfers were mainly chosen according to the donor site of breast reconstruction .Themicrosurgical lymph nodes were transferred to the axillary cavity .When the superficial lymph vessels could be detected in lymphangiography with indocyanine green , thelymphatic-venous anastomoses were done to improve the lymphatic drainage .The treatment effect was assessed by the perimeter changes of different parts of upper extremity, the isotope lymphangiography and associated symptoms . Results 20 cases involved in autologous tissue breast reconstruction with microsurgical lymph node transfers , 18 cases from ingruinallymph nodes and 2 cases from lateral thoracic lymph nodes .2 cases receivedlymphatic-venous anastomoses on their upper extremity .The perimeters of palm and wrist were found significantly decreased in 6 months postoperation , while the perimeters of midpoint forearm and upper arm also decreased .The cellulitis, pain and swell happened less during the follow-up from 6 months up to 4 years. The postoperation isotope lymphangiography showed functional transferred lymph nodes inaxillary cavity , better lymphatic drainage and less volume of upper extremity .The subcutaneous superficial lymphatic drainage signs could be observed by the isotope lymphangiography in cases who had lymphatic -venous anastomoseson upper extremity .Conclusions Autologous tissue breast reconstruction with microsurgical lymph node transfers and lymphatic-venous anastomoses is a promising option for the treatment of mastectomy related axillary cavitydeformation and upper extremity lymphedema .The long term results need longer follow-up and more research .

Objective To investigate the effect of autologous tissue breast reconstruction with microsurgical lymph node transfers and lymphatic-venous anastomoses for the treatment of mastectomy related axillary cavitydeformation and upper extremity lymphedema .Methods The donor sites of lymph node transfers were mainly chosen according to the donor site of breast reconstruction .Themicrosurgical lymph nodes were transferred to the axillary cavity .When the superficial lymph vessels could be detected in lymphangiography with indocyanine green , thelymphatic-venous anastomoses were done to improve the lymphatic drainage .The treatment effect was assessed by the perimeter changes of different parts of upper extremity, the isotope lymphangiography and associated symptoms . Results 20 cases involved in autologous tissue breast reconstruction with microsurgical lymph node transfers , 18 cases from ingruinallymph nodes and 2 cases from lateral thoracic lymph nodes .2 cases receivedlymphatic-venous anastomoses on their upper extremity .The perimeters of palm and wrist were found significantly decreased in 6 months postoperation , while the perimeters of midpoint forearm and upper arm also decreased .The cellulitis, pain and swell happened less during the follow-up from 6 months up to 4 years. The postoperation isotope lymphangiography showed functional transferred lymph nodes inaxillary cavity , better lymphatic drainage and less volume of upper extremity .The subcutaneous superficial lymphatic drainage signs could be observed by the isotope lymphangiography in cases who had lymphatic -venous anastomoseson upper extremity .Conclusions Autologous tissue breast reconstruction with microsurgical lymph node transfers and lymphatic-venous anastomoses is a promising option for the treatment of mastectomy related axillary cavitydeformation and upper extremity lymphedema .The long term results need longer follow-up and more research .

Objective To compare the toxicity of outer membrane vesicles ( OMVs) secreted by Acinetobacter baumannii strains with different drug-resistance spectrums.Methods Four Acinetobacter baumannii strains with different drug-resistance spectrums were collected (strain 33, 3237, B29 and 10), and OMVs produced by these strains were extracted and purified.BCA assay was used to determine the protein concentrations, and RAW264.7 cells were incubated with different concentrations of OMVs for 24 h. Cell viability was measured with CCK-8 assay, and gene expression of tumor necrosis factor-alpha ( TNF-α) , interleukin-6 ( IL-6) , interleukin-1 beta ( IL-1β) , keratinocyte-derived chemokine ( KC) and macrophage inflammatory protein 2 (MIP-2) was assessed by quantitative real-time PCR.One-way ANOVA was used for data analysis.Results According to the result of drug susceptibility test, strain 10 was extensively drug-resistant Acinetobacter baumannii ( XDRAB ) strain, strain B29 was multi-drug resistance Acinetobacter baumannii (MDRAB) strain, while strain 33 and 3237 were non-MDRAB strains.After incubated with different concentrations of OMVs for 24 h, cell viability of RAW264.7 declined with the increase of OMVs concentrations.OMVs released from strain10, B29 and 3237 significantly lowered the cell viability at the concentration of 5 μg/mL, while the cytotoxicity of OMVs released from strain 33 was much weaker, and no remarkable decrease in cell viability was observed even at the concentration of 25 μg/mL.OMVs of all strains induced the release of TNF-α, IL-6, IL-1β, KC and MIP-2 in RAW264.7 cells, and the levels of theses cytokines were increased with the concentration of OMVs.Inflammatory response in cells incubated with OMVs from strain 33 was the weakest, while OMVs from strain 10 induced strongest inflammatory response.KC and MIP-2 levels were significantly higher in RAW264.7 cells incubated with OMVs from strain 10 with a concentration of 5 μg/mL than that incubated with OMVs from other strains ( F=19.094 and 19.032,P<0.05 or <0.01).Conclusions OMVs from Acinetobacter baumannii strains with different drug-resistance spectrums are of different toxicity.OMVs from XDRAB and MDRAB strains have higher toxicities and may induce stronger inflammatory response.

Objective To investigate the effect of mild hypothermia on glucose regulated protein 78 (GRP78) expression in hippocampus following transient cerebral ischemia-reperfusion (I/R) in aged rats.Methods One hundred and forty-four male aged Sprague-Dawley rats,weighing 450-550 g,were randomly divided into 3 groups (n=48 each): sham operation group (group S),I/R group and mild hypothermia group (group H).Global cerebral I/R was induced by 4-vessel occlusion method.Bilateral vertebral arteries were permanently occluded by cauterization,and bilateral carotid arteries were occluded for 5 min.The surface cooling was started immediately after reperfusion and maintained for 3 h.During surface cooling,the body temperature was maintained at 32-34 ℃(rectal).At 6,12,24 and 48 h of reperfusion,the expression of GRP78 was determined using immuno-histochemisty and Western blot,and the viable neurons in CA1 area were counted.Results The number of viable neurons was significantly smaller at each time point,and the expression of GRP78 was significantly higher at 6,12 and 24 h of reperfusion,while lower at 48 h of reperfusion in I/R group,and the number of viable neurons was significantly smaller,and the expression of GRP78 was significantly higher at each time point in group H than in group S (P＜ 0.05).The number of viable neurons was significantly larger at 12,24 and 48 h of reperfusion,and the expression of GRP78 was significantly higher at each time point in group H than in group I/R (P ＜ 0.05).Conclusion Mild hypothermia can further up-regulate GRP78 expression in hippocampus following transient cerebral I/R in aged rats,thus reducing endoplasmic reticulum stress and cerebral I/R injury.