Objective In this study ,we constructed a series of recombinant plasmids carriers expressing shRNA targeting hTERT and Bi‐1 gene .These recombinant plasmids carriers were transfected into CNE‐2Z cell lines using Lip and continuously in‐duced the expression of shRNAs .Furthermore ,the shRNAs caused the degradation of mRNAs homologous in sequence with the target genes ,which lead to a sequence‐specific gene silencing .Methods The CNE‐2Z cells was divided into untreated group ,pEG‐FP‐N1 group and pEGFP‐N1/Lip group .Flow cytometry(FCM ) was applied to determine the transfection efficiency .The changes of hTERT and Bi‐1 gene expression were detected by Real‐time RT‐PCR and Western blotting .Results The best transfection effi‐ciency between plasmid and Lip was 2 .5 μg plasmid and 6 .25μL Lip .Conclusion We constructed several shRNA recombinant eu‐karyotic expression plasmids successfully .The recombinant plasmid can inhibit the expression of hTERT and Bi‐1 gene specifically and effectively .

Objective:To analyze the gene polymorphisms at code 54 of exon 1 in mannose-binding lectin(MBL) gene and the palsma MBL concentrations in Mongols and Huis in China.Methods:MBL plasma concentrations were measured by ELISA method with human MBL ELISA kit,Single nucleotide polymorphisms of MBL gene were determined with sequence analysis method.Geneticanalysis was deployed with SHEsis software,and the statistics package for social science,SPSS11.0 was used in the study.Results:The median values of MBL plasma in 79 Mongols and 69 Huis were 1 686 ng/ml and 1 909 ng/ml respectively and no significant difference was found between them(Z=0.63,P=0.4).In 79 Mongols,the variant allele frequencies of the codon 54 of MBL gene was 0.203,62.0% of the 79 cases were A/A exon type,while 35.4% were A/B type and 2.50% were B/B type instead.In all 69 Huis,the variant allele frequencies was 0.268,58.0% of 40 cases were A/A exon type,while 30.4% were A/B type and 11.6% were B/B type instead.The frequency of point mutation at +230 site was 0.203.No significant difference of variant allele frequencies was found between the Mongols and Huis nationalities(?2=1.772,P=0.183).Neither C nor D exon type were found in the study.The A/A wild type was associated with the highest plasma concentration with median values,the A/B type was associated with lower MBL levels and the B/B type with the lowest levels(?2=86.526,P

AIM:ToinvestigatetheeffectofIkarosisoformsontheproliferationofhumanovariancancerSK-OV3 cells.METHODS:Three isoforms of Ikaros, IK1, IK2 and IK6, were transfected into ovarian cancer SKOV3 cells. CCK-8 assay and cell counting were used to detect the effects of Ikaros isoforms on the proliferation of SKOV 3 cells.The cell cycle was analyzed by flow cytometry .The cell cycle-related proteins were detected by Western blot .RESULTS:IK1 and IK2 expression inhibited SKOV 3 cells proliferation .Flow cytometry analysis indicated that IK 1 and IK2 induced SK-OV3 cell cycle arrest at the G 1 phase.IK6 isoform exerted no obvious effect on the proliferation or cell cycle of SKOV 3 cells.Compared with control EV group , IK1 group and IK2 group showed a dramatic elevation in the expression of the cell cycle inhibitor p21, along with a substantial decrease in the expression of the cell cycle inducers cyclin D 1 and cyclin D2, which did not change in IK 6 group.CONCLUSION:IK1 and IK2 significantly inhibit the proliferation of ovarian cancer SKOV3 cells and induce cell cycle arrest at G 1 phase by regulation of cell cycle-related proteins cyclin D1, cyclin D2 and p21, while IK6 isoform exerts no obvious effect on the proliferation and cell cycle of SKOV 3 cells.

The human cytomegalovirus (HCMV) is a widespread herpesvirus. Virus reactivation from latency can lead to stillbirth, miscarriage, fetal anomalies, and intrauterine growth retardation. During latent infection with the HCMV, the virus can be cleared by the immune response or apoptosis of host cells. However, the HCMV has developed several strategies to manipulate expression of its genes and the microenvironment of host cells. Recent studies have shown that latent infection with the HCMV is associated with viral: regulation of early expression of genes; evasion of cell death; evasion of the immune response; regulatin of non-coding RNAs. This review summarizes recent research progress on the mechanisms underpinning latent infection with the HCMV.
Animals ; Cytomegalovirus ; genetics ; physiology ; Cytomegalovirus Infections ; immunology ; virology ; Humans ; Viral Proteins ; genetics ; metabolism ; Virus Latency

OBJECTIVE: To build information repository of the carrying rate of neonatal deafness gene in Shijiazhuang. METHOD: Blood samples were collected from the heel in 3-days neonates. Mutations of the deafness related genes were detected by the method of fluorescent PCR. Neonates received the detection of 6 mutation sites from 3 genes, including GJB2 (235delC, 299-300delAT), SLC26A4 (IVS7-2A> G, 2168A> G), mitochondrial DNA12S rRNA(1494C>T,1555A>G). RESULT: There were 384 neonates who carried mutations among 421 subjects and the carrying rate was 4.08%, 158 (1.68%) newborns carried heterozygous mutations and 1 (0.01%) case carried homogeneous mutation of GJB2 (235 delC), 55 (0.58%) neonates carried heterozygous mutations of GJB2 (299-300delAT); 133 (1.41%) neonates carried heterozygous mutations and 1 (0.01%) homogeneous of SLC26A4(IVS7-2A>G),19 (0.20%) newborns carried heterozygous mutations of SLC26A4 (2168A>G). The numbers of neonates who carried homogeneous and heterogeneous mutation of mitochondrial 12S rRNA gene were 14 and 3 with carring rates of 0.15% and 0.03%. Two newborns were found to carry more than one mutation. One carried 235delC, IVS7-2A>G and 1555A>G and another carried 235delC and IVS7-2A>G. CONCLUSION The main mutational patterns were 235delC from GJB2 gene and IVS7-2A>G from SLC26A4 gene in Shijiazhuang newborns. The carrying rate information repository of neonatal deafness gene has been built preliminarily.
Connexin 26 ; Connexins ; genetics ; DNA Mutational Analysis ; DNA, Mitochondrial ; genetics ; Deafness ; genetics ; Genetic Testing ; Heterozygote ; Humans ; Infant, Newborn ; Mutation ; Neonatal Screening ; Polymerase Chain Reaction ; RNA, Ribosomal ; genetics

Intracranial infection is a common and serious complication of acute severe traumatic brain injury, with high mortality and disability rates, which significantly affects the prognosis. In recent years, with the widespread use of antibiotics, antibiotic-resistance rates of pathogens have risen year by year, and the choice of sensitive antibiotics is less and less, sometimes even in difficulties of no drugs available. This paper reviewed the treatment process of 1 case with intracranial infection caused by multi drug-resistant Klebsiella pneumonia after severe traumatic brain injury . The aim is to summarize the clinical experience.

Objective To investigate the long-term oncological outcomes of laparoscopic radical hysterectomy (LRH) plus lymph node dissection (LND) and abdominal radical hysterectomy (ARH) plus LND for patients with stage Ⅰ a2-Ⅱ a2 cervical cancer.Methods A retrospective review of stage Ⅰ a2-Ⅱ a2 cervical cancer patients who underwent LRH + LND (n=372) and ARH + LND (n=434) at the First Affiliated Hospital of Sun Yat-sen University from Jan.2005 to Aug.2013 was performed.Individual patient matching was performed by the risk factors for recurrence [tumor size,lymph vascular space invasion (LVSI),depth of cervical stromal invasion,lymph node metastasis,parametrialinvolvement,and resection margin involvement] between two groups.After matched,a total of 203 patient pairs (LRH-ARH) were enrolled.The survival data,surgery data,intraoperative and postoperative complications were compared between the two groups.To assess the prognosis factors,the univariate and multivariate Cox's proportional hazards modelanalysis were conducted.Stratified analysis was performed based on the independent prognosis factors to investigate the survival data between the two surgery groups.Results (1) Surgery data:The operating time [(239±44) vs (270±42) minutes],estimated blood loss [(210± 129) vs (428±320) ml],the duration of bowel motility return [(2.0±0.8) vs (3.0± 1.6) days] and hospital stay [(11 ±6) vs (13±6) days] in the LRH group were significantly shorter than those in ARH group (all P＜0.01).(2) Intraoperative and postoperative complications:The intraoperative complications rate was similar betweentwo groups [6.4％(13/203) vs 6.9％(14/203),P=1.000].The rate of postoperative complications (excluded bladder dysfunction) in the LRH group were significantly lower than those in the ARH group [9.4％ (19/203) vs 20.2％ (41/203),P=0.002].While there was no significant difference in the rates of bladder dysfunction between two groups [36.5％ (74/203) vs 37.4％ (76/203),P=0.910].(3) Recurrence and survival data:There was no significant difference in the recurrence rates between the LRH group and ARH groups [7.9％ (16/203) vs 9.4％ (19/203),P=0.850].There were similar 5-year recurrence-free survival (RFS;92.1％ vs 91.1％,P=0.790) and 5-year overall survival (OS;93.7％ vs 96.1％,P=0.900).(4) Prognosis factor:In univariate analysis,the results showed that tumor size,International Federation of Gynecology and Obstetrics (FIGO) stage,adjuvant therapy,LVSI,stromal invasion,parametrium invasion,pelvic lymph node metastasis,and para-aortic lymph node metastasis were significantly associated with poor prognosis (all P＜0.01).However,age,body mass index (BMI),surgery type,histological type,grade were not significantly associated with poor prognosis (all P＞0.05).The multivariate analysis results,showed that tumor size,pelvic lymph node metastasis,and para-aortic lymph node metastasis were significantly associated with poor prognosis (all P＜0.01).Stratified analysis showed that,even in patients with tumor size ＞4 cm,pelvic lymph node metastasis positive,and para-aortic lymph node metastasis positive in all subgroups,there were not significant difference for the estimated 5-year RFS and 5-year OS between LRH and ARH group (all P＞0.05).Conclusion For patients with stage Ⅰ a2-Ⅱ a2 cervical cancer,LRH plus lymph node dissection is an oncologically safe and surgical feasible alternative to ARH.

Objective To evaluate the efficacy and quality of life of segmental bowel resection for bowel endometriosis. Methods Totally 62 symptomatic patients with bowel endometriosis undergoing segmental bowel resection were recruited. A visual analogue scale (VAS) and the 36-item short form health survey (SF-36) questionnaire were administered before and at least 1 year after surgery, respectively. Pregnancy rates were also recorded. Results Sixty-two patients in total underwent follow-up ranging from 12 to 74 months. All patients complained of obvious pain symptoms, including dysmenorrhea, dyspareunia, pain on defecation and chronic pelvic pain. The relief of dysmenorrhea (2.9 ± 2.2 versus 7.5 ± 2.9), dyspareunia (0.7 ± 0.5 versus 4.3 ± 2.2) and pain on defecation (1.6 ± 0.7 versus 7.3 ± 1.9) after surgery was statistically significant (all P<0.01). The scores for all 8 domains of the SF-36 questionnaire were significant improved after segmental bowel resection (all P<0.01). The complication rate was 45% (28/62), including 18 cases of urinary retention, 4 rectovaginal fistulas, 2 cases of vaginal dehiscence, and 1 case each of thrombogenesis, pelvic abscess and general peritonitis. All of the patients with complications recovered well throughout follow-up. The postoperative pregnancy rate of the previous infertile patients was 6/10. Among the 6 gestational cases, 2 had labour, 2 underwent caesarean sections, one had a spontaneous natural abortion, and one underwent uterine curettage. Conclusion Segmental bowel resection could significantly relieve pain and improve quality of life for patients with bowel endometriosis.

Objective To investigate the effects of saturated fatty acids and unsaturated fatty acids on proliferation and autophagy of human lung cancer cells. Methods The lung cancer cells A549 were treated with stearic acid (saturated fatty acid) and doconexent (DHA, unsaturated fatty acid), respectively, in concentrations of 0, 30, 60, 120 and 240μmol/L. MTT test and cell clone formation assay were performed to detect the proliferation of A549 cells. The morphology of A549 autophagy was observed by confocal laser scanning microscopy after A549 cells were treated with stearic acid or DHA for 24 hours. Western blotting assay was used to detect the expression of autophagy-related protein after A549 cells were treated with stearic acid or DHA for 12, 24 and 36 hours, respectively. Results 30-240μmol/L stearic acid or DHA both inhibited the proliferation of A549 cells (P<0.05). Both stearic acid and DHA induced autophagy of A549 cells, meanwhile, down-regulated Phospho-mTOR (ser2481) and up-regulated LC3Ⅱ/LC3Ⅰ of A549 cells (P<0.05). Conclusions Both saturated fatty acid and unsaturated fatty acid can inhibit the proliferation and induce autophagy of lung cancer cells. The mechanisms of autophagy may be related to Phospho-mTOR (ser2481) signaling pathway.

Objective:To explore the value of high frequency ultrasound combined with serum alpha-fetoprotein (AFP) in the accurate qualitative diagnosis of pediatric testicular tumors.Methods:The ultrasound characteristics (physical properties, calcification, alder blood flow classification) and AFP levels of 47 testicular tumors confirmed by operation and pathology were analyzed retrospectively. The tumors were further divided into two ways: malignant tumor group and benign tumor group, yolk sac tumor group and non yolk sac tumor group. The characteristics of ultrasound and the accurate qualitative diagnosis efficiency of AFP in testicular tumors were analyzed by receiver operating characteristic curve (ROC).Results:18 cases of yolk sac tumor showed solid or almost solid mass, which may be accompanied by several small anechoic areas without calcification. The Alder blood flow grade were grade 3. 29 cases of nonyolk sac tumor showed cystic, solid or mixed mass, most of them have calcification and some of them showed honeycomb echo. Alder blood flow grade were 0-3 grade. ROC curve analysis showed that the area under the curve, sensitivity and specificity of the ultrasound characteristics and AFP in the diagnosis of pediatric testicular malignancies were: solid or almost solid mass (0.894, 83.3%, 95.5%), and no calcification (0.904, 94.4%, 86.4%), Alder blood flow level 3 (0.941, 88.9%, 95.5%), AFP by best cut-off value 18.8 ng/ml (0.972, 100%, 95.5%), ultrasound features combined with AFP (0.992, 100%, 90.9%). All the testicular malignancies, such as yolk sac tumor, immature teratoma, teratoma combined with yolk sac tumor, can be identified by ultrasound features combined with AFP. Further analysis showed that the sensitivity and specificity of the diagnosis of yolk sac tumor with combined solid or almost solid and no calcification were both 100.0%, which can accurately distinguish all cases of yolk sac tumor.Conclusions:Pediatric testicular yolk sac tumor has specific ultrasound performance, high-frequency ultrasound can make a relatively accurate diagnosis, combined with serum AFP can further make a relatively accurate qualitative diagnosis of other malignant tumors of the testis in children.