Objective Two Chinese pedigrees with congenital factor Ⅻ(FⅫ)deficiency were enrolled in the present study,and studies on the clinical manifestations,family survey,biochemical examinations and gene diagnosis of these pedigrees were performed.Methods In October 2014-2015 March,two cases of hereditary FⅫ deficiency patients were included in Xinhua hospital.Activated partial thromboplastin time(APTT),FⅫ procoagulant activity(FⅫ:C),FⅫ antigen(FⅫ:Ag)and other parameters of coagulant were detected.The FⅫ deficiency pedigree members,exons 1-14,boundary introns including the splice junctions of the F12 gene were amplified with polymerase chain reaction(PCR).Direct sequencing was exerted to purified PCR product to detect the gene mutation.If the gene mutations were found,polymorphism should be ruled out by directing sequence.One hundred and three healthy persons as normal controls.Results The two probands were manifested prolonged APTT(101 s and 143 s).They showed lower FⅫ activity and FⅫ antigen(2％and 6％,0.4％and 4％,respectively).FⅡ:C,FⅦ:C,FⅧ:C,FⅨ:C,FⅩ:C and Fg are normal in the two probands.LAC is negative.Proband 1 has c.1285C>T(p.Q429 stop)mutation.His parents and son have the heterozygous mutation in the same position.Proband 2 has c.1556T>C(p.L519P)mutation.Her two sons have the heterozygous mutation in the same position.In the promoter regions of F12 gene,there were common 46C/T and 619 G/C polymorphisms in two pedigrees.Conclusion c.1285C>T(p.Q429 stop)and c.1556T>C(p.L519P)are the cause of FⅫ deficiency.

Objective To study the GPⅡb/Ⅲa gene mutations of eight glanzmann thrombasthenia(GT) pedigrees. Methods Responses of eight probands to different agonists were observed by platelet aggregation test and the amount of αⅡb and β3 was determined by flow cytometry. All the exons of Ⅱb and Ⅲ a genes were amplified by PCR followed by sequencing for mutational screening. Further analysis of the normal population excluded the possibility of mutational sites as a polymorphism. Results Eight probands showed normal PLT counts, dispersion of the platelet particles without aggregation, prolonged bleeding time and severely reduced platelet aggregation in response to the physiological agonists- ADP, epinephrine, and collagen, but relatively normal aggregation of PLT in response to ristocetin. Flow cytometry showed that all probande were Ⅰ type GT, except that proband 2 was Ⅲ type GT and proband 6 was Ⅱ type GT. The sequencing results showed that twelve different types mutations were present in eight probands, including GIOA, Gl412T, GII99A, 1525deiC, G2223T, C2671T, 2930delG, IVSI5 (-1) delG, A2334C, C1750T, 69-79del and CA70A. We were not able to detected any mutations in GP Ⅱb/Ⅲa gone on proband 3. Conclusions GT is mainly caused by GPⅡb/Ⅲa gene mutations. G10A, 69-79del, C470A,G1412T, G2223T, C2671T and 1525delC were the novel mutations causing GT.

Objective To identify the risk factors of Burkholderia cenocepacia associated nosocomial lower respiratory tract infections (NLRTIs),and to investigate the drug resistance of Burkholderia cenocepacia strains.Methods A total of 138 patients with Burkholderia cenocepacia associated NLRTIs and 40 patients with non-Burkholderia cenocepacia associated NLRTIs were enrolled in the study.All patients were collected from the Second Affiliated Hospital of Wenzhou Medical University during January 2009 and December 2012.Clinical data and results of drug sensitivity tests were retrospectively reviewed.Chi-square test and Logistic regression analysis were performed to identify the risk factors of Burkholderia cenocepacia associated NLRTIs.Results Logistic regression analvsis showed that combination use of 2 or more antimicrobial agents,mechanical ventilation,stay in intensive care unit (ICU) for more than two weeks,use of antacid H2 antagonist and deep venous puncture were the independent risk factors of Burkholderia cenocepacia associated NLRTIs (OR =6.315,5.957,5.254,4.585 and 2.017,P ＜0.05).Burkholderia cenocepacia strains were sensitive to levofloxacin,ceftazidime and sulfamethoxazole; More than 40％ strains were resistant to cefotaxime,ceftriaxone,cefepime,aztreonam and tetracycline; And nearly 100％ strains were resistant to gentamicin,amikacin and tobramycin.Conclusion Burkholderia cenocepacia associated NLRTIs are more likely to occur in patients with combination use of 2 or more antimicrobial agents,mechanical ventilation,and those who stay in ICU for more than two weeks,or received antacid and deep venous punctures,and most Burkholderia cenocepacia strains are multiple drug resistant.

Objective To identify the gene mutations of platelet membrane glycoprotein Ⅱ b,Ⅲa(GPⅡb/Ⅲa)in three Chinese pedigrees with Glanzmann thrombastIlenia.Methods All exons and exonintron boundaries of GP Ⅱ b/Ⅲ a gene were amplified by PCR analysis followed by DNA sequencing.DNA sequencing was used to exclude gene polymorphisms.Results The probands in the three pedigrees had a normal platelet count,coagulation profiles,scattered platelets on the blood film,a prolonged cutaneous bleeding time,and impaired or minimal ex vivo platelet aggregation in response to ADP,thrombin,collagen,adrenaline and arachidonic acid,but normal platelet aggregation in response to ristoeetin.Both FACS and Western blotting demonstrated trace content of αⅡb in the platelets from proband 1 and proband 3,who were classified as type Ⅰ GT,and a small amount of αⅡb in the platelets from proband 2,who was classified as type Ⅱ GT.Compound heterozygous mutations,T2255G(Leu721Arg)and C2671T(Gln860Stop)were identified in proband 1.The proband 2 had homozygous A2334C(Gln747Pro)missense mutation.Nonsense mutations C1750T (Arg584Stop)and 69-79 deletion mutation were identified in proband 3. Conclusions Compound heterozygous mutations T2255G and C2671T of αⅡb gene lead to type Ⅰ Glanzmann thrombasthenia for proband 1. Homozygous mutation A2334C of αⅡb gene leads to type Ⅱ Glanzmann thrombasthenia for proband 2. Compound heterozygous mutations C1750T and 69-79del αⅡb gene lead to type Ⅰ Glanzmann thrombasthenia for proband 3. T2255G,C1671T and 69-79del aye novel mutations for αⅡb gene.

Objective To investigate the relationship between the nerve growth factor ( NGF ) induced hippocampal neuroregeneration and homeobox gene Lhx 8.Methods Seventy-two SD rats were divided into control group , transected group, NGF group, transected combined with NGF group after right fimbria-fornix transection and NGF intracerebroventricular injection . Real-time PCR and Western blotting were applied to detect the gene and protein expression of Lhx8 in each group.The choline acetyltransferase ( ChAT)/Lhx8 double labeled cells in subgranular zone ( SGZ) of hippocampus in each group were detected by immunofluorescence .Results The expression of Lhx8 gene and protein in the transected , NGF group and especially in the transected combined with NGF group was obviously higher than in the control group .The number of ChAT/Lhx8 double labeled cells in the NGF group and the transected combined with NGF group was obviously more than in the control group and transected group . Conclusion The hippocampal neuroregeneration which induced by NGF intracerebroventricular injection was associated with the higher expression of Lhx8.

ObjectivesTo detect the effects of P-selectin on deep venous thrombosis (DVT) in nephrotic syndrome (NS). and to evaluate the molecular magnetic resonance imaging (MRI) with a P-selectin targeted conlrost agent in diagnosis of thrombosis in the early phase. Methods(1) Forty-one patients with NS hospitalized in our department from 2005 to 2006 were enrolled in this study. They were assigned into DVT group and non-DVT group according to lower limbs radionuclide imaging (RNV) with 99mTc MAA. Blood P-selectin level was measured by ELISA method. (2) P-selectin was detected both in injured vein and blood immediately, 1 h and 3 h after the dog DVT model was established. (3) The P-selectin-targeted contrast agent was developed by conjugating anti-P-selectin lectin-EGF domain monoclonal antibody (PsL-EGFmAb) which was prepared by our lab. The potential of this contrast agent used in vitro molecular imaging experiment as well as in vivo experiment in dog DVT model was investigated. Results (1) Blood P-selectin level was elevated in patients with NS. It was much higher in DVT group than that in non-DVT group. (2) Blood P-selectin level was also elevated in DVT dogs and P-selectin expressed immediately in tunica intima of injured vein and subsequently in thrombus after the model established. (3) Mural thrombus showed higher signal visualization than surrounding muscle in 30 rain after contrast agent injection. These enhanced signals exhibited P-selectin specificity and persisted from the initiation of intima lesions to 3 h after development of thrombosis. There was signficant Differences in contrast-to-noise ratio (CNR) of the experiment group and the control group (11.50±2.32 vs 2.71±0.86, P＜0.01). The same results were derived from 30 rain to 1 hafter contrast agent being injected in distal to heart part of the injured vessel, and the signal decreased 24 h later. Differences in CNR of the experiment group and the control group were also statistically significant (10.40±2.15 vs 1.93±0.57, P＜0.01). Moreover, the contrast agent did not affect the vital signs of the dog. The function of the heart, lung, liver and kidney functions remained normal after contrast administration. Conclusions P-selectin*targeted new MR contrast can be used to early locate thrombus in vivo in an early stage, which does not compromise the function of the important organs. It may become a new method for early diagnosis of thrombosis.

Objective:To examine the clinical effectiveness of modified vaginal hysterectomy for the treatment elderly women with uterine prolapse.Methods:Eighty-two elderly patients with uterine prolapse treated in the Affiliated Hospital of Jiangnan University from January 2017 to August 2020 were selected as research subjects, who were randomly and double-blindly divided into the control group and the observation group, with 41 patients in each group.The control group was treated with traditional vaginal hysterectomy, while the observation group was treated with modified vaginal hysterectomy.Surgical parameters(operating time, intraoperative blood loss and length of hospital stay), total clinical effectiveness rates and the incidences of complications in the two groups were compared.Results:The total effectiveness rate of the observation group was higher than that of the control group(97.6% vs.78.0%, χ2=7.291; P<0.05). The operation time[(71.3±15.5)min vs.(122.7±22.2)min, t=7.379, P=0.008], intraoperative blood loss[(122.5±15.3)ml vs.(297.7±20.7)ml, t=9.820, P=0.000]and hospitalization time[(7.2±1.5)d vs.(10.0±2.9)d, t=7.164, P=0.014]of the observation group were lower than those of the control group.The incidence of complications in the observation group was lower than that in the control group(0.0% vs.14.6%, χ2=7.159, P<0.05). Conclusions:Modified vaginal hysterectomy for elderly patients with uterine prolapse can help improve the clinical effectiveness and reduce the operating time, postoperative length of hospital stay and the incidence of postoperative complications.

Objective:To investigate the effect of oxycodone hydrochloride injection pretreatment on focal cerebral ischemia-reperfusion injury in rats.Methods:Seventy-two male SD rats weighing 200-250 g were randomly divided into 3 groups( n=24 each group): sham operation group (sham group), focal cerebral I/R group (I/R group), and oxycodone hydrochloride injection group (Oxy group). Focal cerebral I/R was induced by middle cerebral artery occlusion for 2 h followed by reperfusion. In the Oxy group, oxycodone hydrochloride 0.5 mg/kg was injected iv at 5 min before ischemia. While the same volume of saline (1 mL) was injected in the sham group and I/R group. The neurological deficit score (NDS) was assessed at 24 h of reperfusion, the rats were then sacrificed, and their brains were immediately removed for determination of brain water content and the infarct volume, and the histopathological changes were observed after HE staining. The levels of cytokines (TNF-α, IL-1β and IL-10) in the ischemia cortex were quantified by ELISA. MPO activity in the ischemia cortex was assessed. Western blot was used to detect the expression of NF-κB in the ischemia cortex. The data were analyzed using SPSS 20.0 software, multiple-group comparisons were performed using one-way ANOVA, and LSD- t test was used for pairwise comparison between groups. A P<0.05 was considered statistically significant different. Results:Compared with the sham group, NDS, brain water content, relative infarction volume and rate of nerve cell necrosis were significantly increased in the I/R and Oxy groups (all P<0.05). Levels of TNF-α, IL-1β, IL-10, NF-κB and the activities of MPO were increased in the ischemia cortex (all P<0.05). Compared the Oxy group with the I/R group, NDS, brain water content, relative infarction volume and rate of nerve cell necrosis were significantly decreased [(1.7±0.9) vs (2.6±1.1);(79.2±2.4)% vs (84.7±4.2)%; (23.0±5.4)% vs (34.8±6.0)%; (25.2±12.4)% vs (54.8±14.8)%, all P<0.05]. The levels of TNF-α, IL-1β, relative expression of NF-κB, and the activities of MPO were significantly decreased in the ischemia cortex [(4.4±1.2) pg/mg vs (6.5±1.2) pg/mg; (5.4±0.7) pg/mg vs (7.8±0.8) pg/mg; (0.83±0.11) vs (1.23±0.33); (0.27±0.09) U/g vs (0.36±0.14) U/g, all P<0.05] , while the concentration of IL-10 was significantly increased [(20.9±4.5) pg/mg vs (9.2±1.6) pg/mg, t=6.036, P=0.000 1]. Conclusions:Oxycodone hydrochloride can attenuate focal cerebral I/R injury through inhibiting NF-κB activity.

Objective:To examine the efficacy of systematic family intervention (a parent-implemented early start Denver model, P-ESDM) on toddlers at high risk of autism spectrum disorders (IHRASD) who were aged 1 to 2.Methods:The developmental screening for infants aged 1-2 years in Hefei city was performed by using the standardized screening method.The monitoring network of referral-assessment-P-ESDM guidance-follow up-early intervention effect assessment was conducted on the screened children with positive results.A total of 110 patients with IHRASD aged 1 to 2 years were detected.Sixty-three cases that met the inclusion criteria and volunteered to take part in this study were divided P-ESDM group (31 cases) and the control group (32 cases). They were assessed before intervention, 3 and 6 months after intervention separately.Parents of the patients in the P-ESDM group attended 12 weeks of family intervention training courses, 1 hour per week, while parents of the patients in the control group rejected interventions available from us.Results:The proportion of fathers with college degree or above (71.0% vs.43.8%, χ2=7.315, P=0.026) and proportion of mothers with high school/secondary school degree or above (83.9% vs.65.6%, χ2=5.264, P=0.072) were significantly higher in the P-ESDM group than in the control group.Three months after intervention, the P-ESDM group showed decreased Aberrant Behavior Checklist (ABC) scores [29.0(20.0, 45.0) scores vs.48.0(33.0, 50.0) scores, Z=-2.298, P=0.022]and increased Infant-Junior Middle School Social Adaptive Capacity Scale (SM) scores[ 10.0(9.0, 10.0) scores vs.9.0(8.0, 10.0) scores, Z= -2.045, P=0.041], compared with the control group.No significant improvement was found by the Gesell tests in the development quotients (EQs) of five energy areas, namely, the adaptive energy area [(83.86±18.03) scores vs.(75.34±10.49) scores, t=1.734, P=0.090], big movement energy area [(90.24±10.79) scores vs.(85.20±8.97) scores, t=1.595, P=0.118], fine movement energy area [(85.18±14.99) scores vs.(83.41 ± 9.28) scores, t=0.429, P=0.670], language energy area [(59.28±15.01) scores vs.(51.09±9.37) scores, t=1.981, P=0.054] and individual-society energy area [(67.13±14.86) scores vs.(63.50±7.85) scores, t=0.908, P=0.369]. Twelve months after intervention, the P-ESDM group demonstrated significantly decreased ABC scores[20.0(12.0, 33.0) scores vs.45.0(32.3, 52.8) scores, Z=-3.783, P=0.000], increased SM scores[10.0(9.0, 10.0) scores vs.9.0(8.3, 10.0) scores, Z=-2.974, P=0.003], increased EQ of adaptive energy area [(80.83±17.20) scores vs.(72.34 ± 13.18) scores, t=2.203, P= 0.031], increased EQ of the individual-society energy area[(71.87±17.30) scores vs.(62.18±13.91) scores, t= 2.454, P= 0.017]and increased EQ of the language energy area[(68.96±19.93) scores vs.(53.42±14.88) scores, t= 3.515, P= 0.001], compared with the control groups. Conclusions:Early screening, diagnosis, and P-ESDM intervention can improve the outcome of young children aged 1 to 2 years at high risk of ASD.P-ESDM intervention for 12 months demonstrates obvious effects in patients with IHRASD.

Objective: To evaluate the effect of irisin preconditioning on global cerebral ischemia-reperfusion (I/R) injury in rats. Methods: Thirty-six healthy male Sprague-Dawley rats, aged 8-10 weeks, weighing 250-300 g, were divided into 3 groups (n=12 each) using a random number table method: sham operation group (group S), global cerebral I/R group (group I/R) and irisin preconditioning group (group I). Global cerebral I/R was induced by occlusion of bilateral common carotid arteries combined with hypotension (MAP maintained at 35-45 mmHg) in anesthetized rats.At 30 min before ischemia, irisin 10 μg/kg (diluted to 10 μg/ml in normal saline) was intravenously injected in group I, and the equal volume of normal saline was intravenously injected in S and I/R groups.Morris water maze test was performed at day 3 of reperfusion to assess the cognitive function.Rats were sacrificed after the end of morris water maze test, and brains were removed for determination of histopathologic changes in hippocampal CA1 region (using HE staining), neuronal apoptosis in hippocampal CA1 region (Tunel staining), glial fibrillary acidic protein (GFAP) expression (by Western blot), myeloperoxidase (MPO) activity in the hippocampal tissues (by colorimetric assay), and contents of tumor necrosis factor-alpha (TNF-α) and interleukin-1beta (IL-1β) in hippocampal tissues (by enzyme-linked immunosorbent assay). The cell necrosis rate and apoptotic rate were calculated. Results: Compared with group S, the escape latency was significantly prolonged on 1-5 days in group I/R and on 1-3 days in group I, the time of staying at 1st quadrant was significantly shortened, the cell necrosis rate and apoptotic rate were increased, the expression of GFAP was up-regulated, and the activity of MPO and contents of TNF-α and IL-1β were increased in I/R and I groups (P<0.05 or 0.01). Compared with group I/R, the escape latency was significantly shortened on 1-5 days, the time of staying at 1st quadrant was prolonged, the cell necrosis rate and apoptotic rate were decreased, the expression of GFAP was down-regulated, and the MPO activity and contents of TNF-α and IL-1β were decreased in group I (P<0.05 or 0.01). Conclusion Irisin preconditioning can reduce the global cerebral I/R injury in rats, and the mechanism may be related to inhibiting activation of astrocytes in hippocampus and reducing inflammatory responses.