Objective:To analyze the risk factors of bronchopulmonary dysplasia(BPD)in very preterm infants(VPI), and to provide scientific basis for the prevention and treatment of BPD in VPI.Methods:A prospective multicenter study was designed to collect the clinical data of VPI in department of neonatology of 28 hospitals in 7 regions from September 2019 to December 2020.According to the continuous oxygen dependence at 28 days after birth, VPI were divided into non BPD group and BPD group, and the risk factors of BPD in VPI were analyzed.Results:A total of 2 514 cases of VPI including 1 364 cases without BPD and 1 150 cases with BPD were enrolled.The incidence of BPD was 45.7%.The smaller the gestational age and weight, the higher the incidence of BPD( P<0.001). Compared with non BPD group, the average birth age, weight and cesarean section rate in BPD group were lower, and the incidence of male infants, small for gestational age and 5-minute apgar score≤7 were higher( P<0.01). In BPD group, the incidences of neonatal respiratory distress syndrome(NRDS), hemodynamically significant patent ductus arteriosus, retinopathy of prematurity, feeding intolerance, extrauterine growth restriction, grade Ⅲ~Ⅳ intracranial hemorrhage, anemia, early-onset and late-onset sepsis, nosocomial infection, parenteral nutrition-associated cholestasis were higher( P<0.05), the use of pulmonary surfactant(PS), postnatal hormone exposure, anemia and blood transfusion were also higher, and the time of invasive and non-invasive mechanical ventilation, oxygen use and total hospital stay were longer( P<0.001). The time of starting enteral nutrition, cumulative fasting days, days of reaching total enteral nutrition, days of continuous parenteral nutrition, days of reaching 110 kcal/(kg·d) total calorie, days of reaching 110 kcal/(kg·d) oral calorie were longer and the breastfeeding rate was lower in BPD group than those in non BPD group( P<0.001). The cumulative doses of amino acid and fat emulsion during the first week of hospitalization were higher in BPD group( P<0.001). Multivariate Logistic regression analysis showed that NRDS, invasive mechanical ventilation, age of reaching total enteral nutrition, anemia and blood transfusion were the independent risk factors for BPD in VPI, and older gestational age was the protective factor for BPD. Conclusion:Strengthening perinatal management, avoiding premature delivery and severe NRDS, shortening the time of invasive mechanical ventilation, paying attention to enteral nutrition management, reaching whole intestinal feeding as soon as possible, and strictly mastering the indications of blood transfusion are very important to reduce the incidence of BPD in VPI.

Objective To analyze the clinical manifestation and genetic testing in a patient with Adams-Oliver syndrome (AOS) and summarize clinical and genetic characteristics of the dedicator of cytokinesis (DOCK) 6 gene related AOS through reviewing related references.Methods Information of the proband who was hospitalized in Affiliated Children Hospital of Capital Institute of Pediatrics in October 2016 and her family members as well as their DNA samples were collected.The gene sequencing was performed using next generation sequencing technology.Using "Adams-Oliver syndrome"and "DOCK6" as key words,the relevant articles were searched from the Pubmed,China National Knowledge Internet and Wanfang databases and reports of 19 cases were reviewed.Results The proband is an eight months old girl.She presented with severe developmental delay,terminal transverse limb defects and visual loss after birth,and then suffered from tonic seizures and myoclonic seizures at two months old.By physical examination she was found to have esotropia and visual loss.The distal phalanx and nail of the right second-fourth fingers were absent,while the phalangette of the left second-fourth fingers and bilateral distal phalanges of toes were short with small nails attachment.Thyroid function test showed hypothyroidism.The ocular fundus examination showed the residual vitreous artery in the left eye and the retinal pigment degeneration in the right eye.CT scan showed multiple bilateral periventricular calcification and cranial magnetic resonance imaging showed bilateral periventricular lesion.Two heterozygous mutations were identified in DOCK6 gene:one was a known pathogenic mutation (p.L1064Vfs*60),and the other was a novel splice site mutation (c.873+ 1G＞A).By analyzing this case and reported 19 cases,the common performances of DOCK6 gene related AOS included terminal transverse limb defects (20/20),aplasia cutis congenita (18/20),ocular abnormalities (13/20),seizures (12/20),mental retardation (12/20),microcephaly (10/20),cardiovascular malformations (10/20),intrauterine growth retardation (6/20).The mutation of the DOCK6 gene was found to be dominated by frameshift mutation and splice site mutation.Conclusions If either terminal transverse limb defects or aplasia cutis congenita was detected in a patient,AOS should be under consideration.In addition,autosomal recessive inheritance,nervous system and eyes involvement will further indicate DOCK6 gene related AOS.

Objective To explore the effectiveness and compliance of ketogenic-diet(KD) treatment for infantile spasm(IS).Methods Ninety-eight IS patients who were treated with KD in Wuhan Children's Hospital from March 2009 to June 2015 were analyzed by using retrospective case-control study,the patients were divided into 4 groups:newly diagnosed IS patients group (group A,including 24 patients),one antiepileptic drug (AEDs) failure IS patients (group B,including 28 patients),two and more AEDs failure IS patients (group C,including 29 patients),and two or more AEDs combined with ACTH failure IS patients(group D,including 17 patients).The spasm-free andretention rates after 3,6 and 12 months KD treatment were compared among these groups.Results Overall retention rate was 80.6％ (79/98 cases),69.4％ (68/98 cases),and 42.9％ (42/98 cases)at 3,6,12 months,respectively.The 3-month retention rate in group A,B,C and D was 83.3 ％ (20/24 cases),78.6％ (22/28 cases),82.7％ (24/29 cases) and 76.4％ (13/17 cases) respectively,and there was no significant difference among these groups (P ＞ 0.05).The 6-month retention rates in each group was 75.0％ (18/24 cases),67.9％ (19/28 cases),68.8％ (20/29 cases) and 65.0％ (11/17 cases) in sequence,and there was also no significant difference among these groups(P ＞0.05).The 12-month retention rate was 54.2％ (13/24 cases),21.4％ (6/28 cases),48.3％ (14/29 cases) and 52.9％ (9/17 cases) in group A,B,C and D in sequence,the 12-month retention rate of group B was significantly lower than that of other 3 groups,and the differences were statistically significant(x2 =5.973,4.508,4.727,all P ＜ 0.05),and there was no significant difference among the A,C,D groups (all P ＞ 0.05).The spasm-free rate at 3,6,12 months of KD treatment was 19.4％ (19/98 cases),20.4％ (20/98 cases),30.6％ (30/98 cases).The 3-month spasm-free rate in A,B,C,D groups were as follow:41.7％ (10/24 cases),14.3％ (4/28 cases),10.3％ (3/29 cases),11.8％ (2/17 cases),respectively.The 3-month spasm-free rate in group A was significantly higher than that of other 3 groups,and the differences were statistically significant (x2 =10.238,9.219,6.697,all P ＜ 0.05),but there was no significant difference among the B,C,D groups (all P ＞ 0.05).The 6-month spasm-free rates were 41.7％ (10/24 cases),14.3％ (4/28 cases),13.8％ (4/29 cases),and 11.8％ (2/17 cases) in group A,B,C and D in order,and the spasm-free rate in group A was significantly higher than that of other 3 groups,and the differences were statistically significant(x2 =4.924,5.249,4.298,all P ＜ 0.05),but there was no significant difference among the A,C,D groups (all P ＞ 0.05).The 12-month spasm-free rates were 54.2％ (13/24 cases),21.4％ (6/28 cases),24.1％ (7/29 cases),and 23.5 ％ (4/17 cases) in group A,B,C and D,and the spasm-free rate in group A was significantly higher than that in other 3 groups,and the differences were statistically significant(x2 =8.354,7.923,4.364,all P ＜ 0.05),but there was no significant difference among the A,C,D groups (all P ＞ 0.05).Conclusions The spasm-free rate of KD therapy for newly-diagnosed IS is higher than that of IS patients whose drug-therapy failed.KD therapy may be the top priority for IS patients and part of those patients whose drug-therapy failed can still get seizure-free with KD diet.

Objective To assess the effectiveness and safety of dexmedetomidine (Dex) in anesthesia for end-stage renal disease (ESRD)recipients in living related renal transplantation.Methods Forty ESRD patients undergoing living related renal transplantation in the Affiliated Provincial Hospital of Anhui Medical University from October 201 3 to December 201 4 were randomized into the Dex group and the control group,20 patients in each group.In the Dex group,the patients were pumped with the loading dose of Dex at 0.6 μg/kg before anesthesia induction and the procedure was completed within 20 min.Then,the patients were pumped at 0.2 μg/(kg·h)for 1 h and underwent general anesthesia induction.In the control group,the patients were pumped with equivalent normal saline.The anesthesia induction and the maintenance drug of the two groups were the same.The mean arterial pressure (MAP),heart rate (HR)and bispectral index (BIS) before administration (T0 ),20 min after administration (T1 ),before laryngeal mask placement (T2 )and after laryngeal mask placement (T3 )of the two groups were observed and recorded.Delayed recovery and emergence delirium were also observed.Urine output during transplantation,4,8,1 2,24 h after transplantation,as well as endogenous creatinine clearance rate (Ccr)before transplantation and 1 2,24 h after transplantation were recorded.Results Compared with T0 ,the MAP,HR and BIS of the two groups at T2 decreased significantly (all in P <0.05).As for the comparison between two groups,the HR and BIS of the Dex group decreased significantly (both in P <0.05 ),but the MAP didn't decrease significantly (P >0.05 ).The emergence delirium in the Dex group was all mild with the incidence of 1 5% (3 /20),which significantly deceased in comparison with that in the control group (30%,6 /20)(P <0.05).The urine outputs of Dex group during transplantation,and 4 h and 8 h after transplantation were significantly higher than those of the control group (all in P <0.05).The Ccr of the two groups at 1 2 h and 24 h after transplantation significantly increased, compared with that before transplantation (all in P <0.01 ).Conclusions Dex may reduce the incidence of emergence delirium of recipient in living related renal transplantation,increase urine output after transplantation and cause no delayed recovery,which may be used in ESRD patients safely.

Objective To evaluate the clinical effects and experiences of short stem ceramic-on-ceramic total hip arthroplasty in the treatment of young and middle-aged patients with femoral head necrosis.Methods From December 2009 to March 2011,40 patients with femoral head necrosis were treated primarily with short stem ceramic-on-ceramic hip arthroplasty (43 hips) through posterior-lateral or anterior-lateral approach.There were 23 males,17 females； with the mean age of 40.5 years (range,27-55 years).All cases were stage Ⅳ or more according Steinberg stage.The patients were followed up at the 3,6,12 months after operation,and then once each year.Standard X-ray films were taken at the follow-ups.Femoral prosthesis subsidence was assessed through X-ray films.Harris scores system and WOMAC Osteoarthritis Index were recorded before and after operation to evaluate clinical effects of surgery.Results Thirty-eight patients (41 hips) were followed up for 21-36 months,with an average of 31.3 months.The femoral stem subsided 0.68±2.90 mm at the last follow-up.After operation,hip pain of patients was relieved obviously.The Harris score improved from 38.7±12.1 (15-66) preoperatively to 95.2±4.49 (83-100) postoperatively,WOMAC Index from 66.7±12.0 (49-94) to 12.8±5.6 (0-24).During follow-up period,there was no serious complication such as femoral neck fracture,infection of wound,pulmonary embolism,et al.No dislocation,implant prosthesis failure and infection were observed.Conclusion The short stem hip arthroplasty preserves bone stock to a great extent.The short stem implant showed a good stability and osseous integration.The wear rate of ceramic-on-ceramic bearing surface is extremely low.Short stem hip arthroplasty plus ceramic-on-ceramic bearing surface might be a good option for the young and middle-aged patients with femoral head necrosis.

ObjectiveTo explore the expression of novel protein kinase C ε (PKCε) in normal prostate (NP) tissue, benign prostate hyperplasia(BPH), peficancerous (PC) tissue and prostate cancer (Pca), and study its correlation with the grade and stage of Pca.MethodsTen NP slides, ten BPH slides, ten PC slides and 43 Pca slides were collected from our hospital. These slides were routinely proceased and analyzed according to the requirement of immunohistochemical staining. Tumors were classified according to the 2002 TNM staging system. The grading system used in the study was based on the Gleason grade.ResultsWe was found that the expression of PKCεs in Pca (27/43) were significantly higher than those in NP(1/10), BPH (0/10) and PC (2/10) tissue, and the difference was statistically significant ( P ＜0.05 ). With regard to grade of prostate cancer, the expression of PKCε in Pca with Gleason score ≥8 group (12/13) was higher than the Gleason score 2 -4 group (4/10) and the Gleason score 5 -7 group (11/20). The difference was statistically significant (P ＜ 0.05 ). Moreover, the T3 and T4 stages had a more positive rate (10/12 & 9/10) than the T1 and T2 stages( 1/6 &7/15). There is statistically significant difference between early and advanced stage of prostate cancer ( P ＜ 0. 05 ). Furthermore, the positive expression of PKCε in prostatic carcinoma samples increased significantly in the metastasis group (9/10)compared to the non-metastasis group ( 18/33 ) ( P ＜ 0. 05 ), but the difference was not statistically significant between the concentration of prostate-specific antigen in blood serum ( P ＞ 0. 05 ).Conclusions PKCε is expressed in prostate cancer, and it correlates with the grade and stage of prostate cancer. PKCε may be related to the origin and the development of Pca, and it may be used as a prognostic factor for Pca.

The present study examined the role of Wnt/β-catenin signaling pathway in the degeneration of nucleus pulposus cells and the protective effect of DKK1 on nucleus pulposus cells. The model of nucleus pulposus cell degeneration was induced by intra-disc injection of TNF-α, and the expression of β-catenin protein was detected by Western blotting. The cultured rabbit nucleus pulposus cells were divided into 4 groups. In group A, the cells were cultured with normal medium and served as control group. In group B, the cells were cultured with TNF-α and acted as degeneration group. In group C, the cells were cultured with TNF-α and transfected with Adv-eGFP and was used as fluorescence control group. In group D, the cells were cultured with TNF-α and transfected with Adv-hDKK1-eGFP, serving as intervention group. The expression of type II collagen, proteoglycan, β-catenin, and MMP-13 in each group was detected by immunocytochemistry and RT-PCR. The result showed that TNF-α increased the expression of β-catenin and MMP-13, and significantly inhibited the synthesis of type II collagen and proteoglycan, which resulted in the degeneration of nucleus pulposus cells. This effect could be obviously reversed by DKK1. We are led to concluded that TNF-α could activate the Wnt/β-catenin signaling pathway, and increase the expression of MMP-13, thereby resulting in disc degeneration. Specifically blocking Wnt/β-catenin signaling pathway by DKK-1 could protect the normal metabolism of intervertebral disc tissue. The Wnt pathway plays an important role in the progression of the intervertebral disc degeneration.

The present study examined the role of Wnt/β-catenin signaling pathway in the degeneration of nucleus pulposus cells and the protective effect of DKK1 on nucleus pulposus cells.The model of nucleus pulposus cell degeneration was induced by intra-disc injection of TNF-α,and the expression of β-catenin protein was detected by Western blotting.The cultured rabbit nucleus pulposus cells were divided into 4 groups.In group A,the cells were cultured with normal medium and served as control group.In group B,the cells were cultured with TNF-α and acted as degeneration group.In group C,the cells were cultured with TNF-α and transfected with Adv-eGFP and was used as fluorescence control group.In group D,the cells were cultured with TNF-α and transfected with Adv-hDKK1-eGFP,serving as intervention group.The expression of type Ⅱ collagen,proteoglycan,β-catenin,and MMP-13 in each group was detected by immunocytochemistry and RT-PCR.The result showed that TNF-α increased the expression of β-catenin and MMP-13,and significantly inhibited the synthesis of type Ⅱ collagen and proteoglycan,which resulted in the degeneration of nucleus pulposus cells.This effect could be obviously reversed by DKK1.We are led to concluded that TNF-α could activate the Wnt/β-catenin signaling pathway,and increase the expression of MMP-13,thereby resulting in disc degeneration.Specifically blocking Wnt/β-catenin signaling pathway by DKK-1 could protect the normal metabolism of intervertebral disc tissue.The Wnt pathway plays an important role in the progression of the intervertebral disc degeneration.

This study sought to construct recombinant eukaryotic plasmid pcDNA3. 1-sFlt-1 and observe its effect on proliferation of vascular endothelial cells. Total RNA was extracted from human umbilical vein endothelial cells (HUVECs) firstly. The 1st-3rd Ig-like domains of Flt were amplified by polymerase chain reaction (PCR) from the full-length cDNA. Subsequently, the PCR product was cloned into the eukaryotic plasmid pcDNA3.l(+)/myc-His. The constructed recombinant plasmid pcDNA3. 1-sFlt-1 was sequenced. Then recombinant plasmid was transfected into Lewis lung cancer cells. RT-PCR and SDS-PAGE were used to detect the expression of soluble vascular endothelial growth factor (VEGF) receptor mRNA and protein, respectively. MTT method was used to evaluate the effect of sFlt-1 protein on proliferation of HUVECs induced by VEGF. The results showed: (1) The sequence of inserted fragment was correct. (2) Lewis lung cancer cells with recombinant plasmid transfection could express the soluble VEGF receptor mRNA and protein stably. (3) Culture supernatant of Lewis lung cancer cells with sFlt-1 could significantly inhibit the proliferation of HUVECs induced by VEGF. These data suggested that recombinant eukaryotic plasmids pcDNA3. 1-sFlt-1 was constructed successfully, sFlt-1 mRNA and protein were expressed in eukaryotic system stably and sFlt-1 protein could significantly inhibit the proliferaton of endothelial cells induced by VEGF.
Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cells, Cultured ; Genetic Vectors ; genetics ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Transfection ; Vascular Endothelial Growth Factor A ; pharmacology ; Vascular Endothelial Growth Factor Receptor-1 ; biosynthesis ; genetics ; pharmacology