Objective:To explore the genetic etiology of a child with Cowden syndrome 1 (CS1).Methods:A child who had visited the Ningbo Women and Children's Hospital on August 26, 2022 was selected as the study subject. Clinical information of the child was collected. Genomic DNA was extracted from peripheral blood samples of the child and his family members and subjected to whole exome sequencing (WES). Candidate variant was verified by Sanger sequencing.Results:The child, a 13-year-old boy, had manifested with severe mental retardation, hyperactivity, autistic behavior, sparse and prominent teeth, macrocephaly, and skin freckles on the penis. His mother had presented with multiple papules, hamartomatous polyps, thyroid adenoma and macrocephaly. WES results revealed that the child has harbored a nonsense c. 781C>T (p.Q261*) variant of the PTEN gene, which was inherited from his mother. Based on the guidelines from the American College of Medical Genetics and Genomics, the c.781C>T variant was classified as likely pathogenic (PVS1+ PM2_Supporting). Conclusion:The c. 781C>T variant of the PTEN gene probably underlay the pathogenesis in the child and his mother. Above finding has facilitated genetic counseling for this family.

Objective To evaluate the clinical value of apheresis platelets throught heanalysisof case control on the clinical efficacy and safety of cryopreserved apheresis platelets and fresh apheresis platelets.Methods 2 035 clinical cases of platelet transfusion in August 2014 to December 2016 by Using the closed loop intelligent path management and evaluation information system,456 cases were selected as control cases.Platelets were divided into the cryopreserved apheresis platelets group (group A,n=199) and fresh apheresis platelets group (group B,n=257) according to the transfused platelet type.The clinical application value of cryopreserved single platelets was evaluated by comparing the basic data,the effective indexes and safety indexes of the two groups.Results 1) The cases were 43.6％ (199/456) in A groups,and 56.4％ (257/456) in B groups,there were no significant difference in gender,age and medical and surgical cases between A and B group (P＞0.05);2) 199 cases in group A were cryopreserved platelets of 2 275 U,including 121 medicine cases,the total amount of transfusion was about 60.9％ (1 385/2 275),78 surgical cases accounted for 39.1％ (890/2 275);In the distribution of diseases,the blood system diseases accounted for 49.2％ (1 120/2 275),the total amount of obstetrics and gynecology disease infusion accounted for 10.6％ (240/2 275),and the amount of tumor radiotherapy and chemotherapy accounted for 6.2％ (140/2 275);The proportion of ABO blood type distribution was O type 25.9％,A type 22.9％,Btype 20.7％,ABtype 30.5％,respectively;3) The Plt counts of group A and B were significantly different before and after transfusion (P ＜0.05).But there was no significant difference between the two groups of cases before transfusion and 24h Plt count after transfusion,the Plt counts difference,and 24 h CCI (P＞0.05);4) The effective rates of platelet transfusion in group A and B were 76.9％ and 76.7％,respectively.Which has no significant difference between the two groups (P＞0.05)).There was no significant difference between the two groups in medical and surgical cases (P＞0.05),but the effective rate of surgical cases in group A (84.6％) was higher than that in B group (75.3％).The difference effect of medicine and surgery cases in B group were not statistically significant (P＞0.05),but the difference effect of medicine and surgery cases in A group was statistically significant (P＜0.05),platelet transfusion inefficient in surgical cases (15.4％) was significantly lower than that of cases (28.1％);5) The incidence of adverse reactions of blood transfusion was 3.5％,4.7％ in group A and B,and the blood transfusion mortality rate was zero,the difference was not statistically significant (P＞ 0.05).Conclusion The clinical effectiveness and safety of cryopreserved apheresis platelets are similar to those of fresh apheresis platelets,and the former can be widely Used in clinic,in particular,it has certain advantages in the surgical hemostatic effect.but for repeated infusion cases or platelet transfusion ineffective cases should be given priority to fresh apheresis platelets.

Objective To analyze the influence of different factors and their relating correlation results on platelet transfusion during the bone marrow empty window period on the patients who have undergone allogeneic peripheral blood stem cell transplantation (allo-HSCT) with retrospective analysis of case-control data.Methods Clinical data of 153 cases were collected by the clinical blood management and evaluation information system with discharge diagnosis of allo-HSCT in the hematology department of The First Affiliated Hospital of Nanchang University within a time frame from January 2014 to December 2016.A total of 90 cases were considered valid for retrospective analysis according to the case exclusion criteria.The average transfusion dose for patients with allo-HSCT during the bone marrow empty window period was defined as the threshold value which divided the 90 cases into the observation group of 38 cases receiving more than 6 Units of platelet transfusion and the control group of 52 cases with less than 6Units of platelet transfusion.The amount of platelets transfused during the bone marrow empty window period,clinical indexes include Hb,ANC,Plt,SF before pretreatment,platelet engraftment time and the number of mononuclear cells implanted were compared and analyzed by Logistic regression.Results (1) There was no significant difference between the two groups in gender,age,primary diagnosis,HLA matching,Hb before pretreatment and the number of mononuclear cells implanted (P＞0.05).The ANC(×109/L) (1.24±0.57 vs 3.36±1.33) and Plt(×109/L) (43.55±68.29 vs 126.62±84.73) counts before pretreatment in the observation group were significantly lower than those in the control group(P＜0.05).SF(μg/L) (2351.05 ± 1 587.96 vs 1 000.96± 362.97)before pretreatment and P LT recovery time (d) (16.84± 2.47 vs 12.73 ± 1.65)was significantly higher than that in the control group(P＜0.05).Donor-recipient ABO blood group typing incompatibility (15 vs 10) was significantly higher than the control group (P＜0.05);(2) Single factor Logistic regression analysis showed that ABO blood group matching,clinical indexes include ANC,Plt,SF before pretreatment,PLT recovery time were statistically significant,Only ANC before pretreatment and PLT recovery time had significant effect on the platelet transfusion during bone marrow empty window period in patients with allo-HSCT in multivariate Logistic regression analysis(P＜0.05).Condusion The ANC before pretreatment and PLT recovery time are independent factors for platelet transfusion of the bone marrow empty window period in patients with allo-HSCT.The PLT recovery time is an independent risk factor,which indicates that the longer the duration of PLT implantation,the greater the amount of platelet transfusion will be needed.Besides,the ANC before pretreatment is the independent protective factor,which indicates that the greater the ANC,the smaller the amount of platelet transfusion is required.

Objective To describe the clinical characteristics and outcomes of dengue virus infection during pregnancy . Methods A retrospective study was performed based on the clinical data of manifestations ,pregnancy outcomes and neonates in patients with dengue virus infection during pregnancy from January 1st ,2014 to December 31th ,2014 at maternity ward in Guangzhou .Results The most common manifestations in patients with dengue virus infection during pregnancy were fever (100 .0％ ) , body soreness (42 .9％ ) ,headache (41 .3％ ) ,cutaneous rash (41 .3％ ) ,fatigue (34 .9％ ) and muscle pain or arthralgia (17 .5％ ) , nausea and vomiting (12 .7％ ) . The decline proportions of leukocyte , platelet and neutrophil count were 17 .5％ ,20 .6％ and 6 .3％ ,respectively ,and the lowest point was observed on 5 — 6 days after the onset of illness .Miscarriage was observed in 3 pregnant patients (4 .8％ , all in the first trimester ) ,intra-uterine death in one patient (1 .6％ ,at 22 weeks of gestation) and artificial abortion in 2 cases .A total of 48 patients delivered ,among whom caesarean section rate was 37 .5％ , prematurity rate was 12 .5％ and low birth weight rate was 8 .2％ .Maternal – foetal vertical transmission happened in 2 cases .Sequence alignment demonstrated that the homologies of the nucleic acids and amino acids within each dengue virus vertical transmission mother-infant pairs were > 99 .9％ .Conclusions The manifestations of dengue virus infected women with pregnancy are not specific .Early detection and early diagnosis should be made according to the history of epidemiology for women with reproductive age in endemic areas .Maternal dengue virus infection during pregnancy might cause vertical transmission and is correlated with bad outcomes of pregnancy , including miscarriage , intra-uterine death and premature birth .Perinatal infection might cause maternal-foetal vertical transmission .

Objective To investigate the effects of extended nursing based on relationship, information and management on the self-care behaviors of patients with heart valve replacement. Methods According to admission time, 37 patients with heart valve replacement were set as the control group, which was given routine nursing intervention and another 42 patients as the study group, which received extended nursing based on the trinity of relationship, information and management 2~8 weeks after discharge. Before and after intervention, patients were evaluated by using the improved version of self-care behavior evaluation scale of heart disease (SNBAOHD). Result There were significant differences between the two groups in other items of SNBAOHD, except two items of low salt diet and medication (P<0.05). Conclusions Extended nursing after discharge based on the trinity of relationship, information and management can help the patients to be continuously cared. In this case, it is effective for the enhancement of off-hospital self-care ability and prevention of disease relapses.

Objective To construct and express a prokaryotic expression vector carrying the gene of FimH 1-156 that comprises human lysosome membrane protein 2 P41-49 gene ,and to express and purify the fusion protein .Methods FimH1-156 gene was cloned from plasmid pPKL241 by PCR ,and inserted into vector pET-28a(+ ) to obtain prokaryotic expression plasmid pET-28a-FimH . After transforming Escherichia coli BL21(DE3) with pET-28a-FimH ,fusion protein FimH1-156 was expressed under induction .The target fusion protein was purified ,and its antigenicity was detected through Western blot .Results The expressed recombinant pro-tein was purified ,the expression of protein was the highest when IPTG was 1 mmol/L and 4h after induction ,it was expressed as include body form ,and the expressed protein was identified to react with monoclonal antibodies 6 × His by Western blotting .Conclu-sion We cloned FimH1-156 fusion protein expressed genes successfully ,constructed prokaryotic expression vector ,and won the in-clusion body purification of FimH1-156 fusion protein .

Objective To evaluate the safety and clinical efficacy of stent implantation using a rapid artificial cardiac pacing technique on ostial lesions of left anterior descending artery (LAD).Methods From Jun 2008 to Nov 2010,38 patients with ostial lesions of LAD were recruited and randomly divided into two groups:patients with stent implantation using a rapid artificial cardiac pacing technique (pacing group,n =19 ) and patients with direct stent implantation (no-pacing group,n =19 ).Post-stenting examination was performed.Patients were followed-up for 9 months and coronary angiography was reviewed.The immediate success rate,major adverse cardiac events including death,reinfarction and target vessel revascularization,late lumen loss,sent thmmbosisin,rent-restenosis were compared between these two groups.Results There were no significant differences in the baseline values,disease characteristics and instant response to surgery between pacing and no-pacing groups(P ＞ 0.05 ).The time cost for stent placement was significantly shorter in the pacing group than that in the no-pacing group ( [ 16.5 ± 0.5 ] s vs.[46.6 ± 1.4 ] s,t =88.256,P =0.004 ).After surgery,there was no acute or subacute thrombosis,in-stent restenosis or occlusion for patients in the pacing group.In the no-pacing group,one patient developed acute thrombosis.The symptoms disappeared after thrombus aspiration and balloon dilatation by emergency percutaneous coronary intervention.Patients were followed up for 270 -275 days,and patients in the pacing group received post-stenting coronary angiography 9 months after stent implantation using a rapid artificial cardiac pacing technique,and no in-stent re-stenosis was found.During this period,nobody had adverse events such as death,myocardial infarction or target lesion revascularization,while five cases with in-stent restenosis (50％ -60％ )were found in the no-pacing group,without further target lesion reconstruction due to symptomless.Conclusion Compared with previous positioning technique,stent implantation using a rapid artificial cardiac pacing technique on treatment of ostial lesions of LAD is safer,and more effective.It is a favorable method for accurate positioning of bracket and can improve the prognosis,reduce the occurrence of acute thrombosis and in-stent restenosis.

The curriculum system of Proteomics was analyzed based on the teaching practice,the characteristics of ability training and gradation teaching were summarized and the prospect of curriculum optimization was proposed.These measures were conceived to enrich the course content and teaching methods for Proteomics course.

Objective To explore how to improve the immunogenicity of short epitope peptides of triggering melanoma MART 1 specific CD8 +T cell responses Methods Therapeutic peptides based on the immunodominant MART 127 35, HIV Tat49 57CCP sequence and a tetanus toxoid universal Th epitope were designed and synthesized The immunological functions were studied in PBMCs from HLA A2 + melanoma patients Results The results demonstrated that the peptides could trigger vigorous MART 1 specific CD8 + CTL activities in vitro The function of peptide containing MART 127 35 and tetanus universal Th epitope was more vigorous than that of MART 127 35 peptide, and the immunogenicty of the peptides with HIV Tat49 57CCP sequence, MART 127 35 and tetanus universal Th epitope was the most vigorous Conclusion Linkage of HIV Tat49 57CCP sequence and a tetanus universal Th epitope could dramatically improve the immunogenictiy of the MART 127 35 epitope peptide

Objective To develop new methods to cultivate, retrieve and purify mouse mesenchymal stem cells (mMSCs). Methods Bone marrow was collected from 2-month-old Kunming mice by flushing femurs and tibias with complete medium of DMEM-LG. Cells were plated in a Petri dish. After 24 hours, non-adherent cells were removed by two to three washes with PBS, adherent cells were further cultured in complete medium and retrieved by trypsinisation with 0.25% trypsin for 5 min at 37 ℃. The treated adherent cells were cultivated with 3?dilution for further generations. CD11b-negative cells were retrieved from the collected adherent cells of 3rd generation by using immunomagnetic microbeads, and continued to be cultured in complete medium. After the cultured cells were retrieved, their morphology and their ability of osteoblastic differentiation and adipocytic differentiation were examined. Results Most of mMSCs from 1st generation were of shuttle shape, some of irregular shape. After treatment with magnetic microbeads and several generations, mMSCs were of spindle, star and irregular shape. These cells were of rich cytoplasma, clear nucleolus, and grew in parallel or vortex. The cultured adherent cells from the first and subsequent generations had plenty of CD11b-positive blooding-making cells. After 20-day osteoblastic induction, mMSCs differentiated into bone cells, which showed orange phosphate in extracellular matrix by Alizarin red S staining. mMSCs could differentiate into lipocytes. The size of cells increased along with fat-developing induction period. These cells showed many orange fatty follicles with O Red Oil dyeing. Conclusion Pure mMSCs can not be retrieved by either adhering method or generation cultivation method separately. The combined methods of adhering, immunomagnetic microbeads, and serial subcultivation is effective in vitro in retrieve mMSCs.