Objective： This study was designed to investigate the relationship between p16 protein expression and gastric carcinogenesis,depth of invasion, lymph node metastasis, and evaluate the role of deletion and mutation of p16 gene in exon 2 in gastric carcinoma. Methods： p16 protein expression in gastric carcinoma and precancerous lesion was examined by streptavidin-peroxidase conjugated(S-P) method; The deletion and mutation of p16 gene were examined respectively by polymerase chain reaction(PCR) and polymerase chain reaction single-strand conformation polymorphism analysis(PCR-SSCP) in gastric carcinoma. Results： ① The positive rates of p16 protein expression were 96.25％ (77/80) in normal gastric mucosa, 92.00％ (45/50) in dysplastic gastric mucosa, and 47.54％ (58/122) in gastric carcinoma. The positive rate of p16 protein expression in gastric carcinoma was significantly lower than that in normal gastric mucosa and in dysplastic gastric mucosa (P<0.05). ② The positive rate of p16 protein expression in mucoid carcinoma (10.00％ ,1/10) was significantly lower than that of poorly differentiated carcinoma (51.22％ ,21/41), undifferentiated carcinoma (57.69％ ,15/26), and signet ring cell carcinoma (62.50％ ,10/16) (P< 0.05). ③ The positive rates of p16 protein in 30 cases paired primary and lymph node metastatic gastric carcinoma were 46.67 ％ (14/30) in primary gastric carcinoma,16.67％ (5/30) in lymph node metastatic gastric carcinoma. The positive rate of lymph node metastatic carcinoma was significantly lower than that of primary carcinoma(P<0.05). ④ Evaluation of mutation and deletion of p16 gene： There was no mutation of p16 gene in exon 2, but there were 5 cases displayed deletion of p16 gene in exon 2 in the 25 primary gastric carcinoma. Conclusions： ① The expression loss of p16 protein is related to carcinogenesis, histopathological subtypes,and lymph metastasis of gastric carcinoma. ② The mutation of p16 gene in exon 2 may not be involved in gastric carcinogenesis. But the deletion of p16 gene in exon 2 might be involved in gastric carcinogenesis.

OBJECTIVETo observe the effect of combined therapy of Neiyi pill (NP) and Neiyi enema (NE) on endometriosis and its effect on serum levels of endometrial antibody (EMAB) and carcinoembryonic antigen 125 (CA125).

METHODSFifty-eight cases with endometriosis were divided into 3 groups randomly, group A (n = 16) treated by NP, group B (n = 24) treated by NP and NE, and group C (n = 18) treated by danazol, all for 3 menstrual cycle with single blind method. The effect was observed and the serum levels of EMAB and CA125 were detected before and after treatment.

RESULTSComparison of the efficacy between the 3 groups showed: there was no remarkable difference between group A and B (P >0.05), both of group A and group B were superior to that of group C (P<0.05). The levels of EMAB and CA125 had no significant changes in all the three groups after treatment.

CONCLUSIONCombined therapy of NP and NE could improve the curative effect on endometriosis, and without obvious effects on serum levels of EMAB and CA125.

Administration, Rectal ; Adult ; Autoantibodies ; blood ; CA-125 Antigen ; blood ; Drugs, Chinese Herbal ; administration & dosage ; Endometriosis ; blood ; drug therapy ; Endometrium ; immunology ; Enema ; Female ; Humans ; Phytotherapy ; Single-Blind Method ; Tablets

Objective To explore the mutation of androgen receptor(AR)gene in a patient with 46,XY disorder of sex development(DSD)and to improve the diagnostic level and understanding of androgen insensitivity syndrome(AIS).Methods The clinical data of the child was analyzed,including physical examination,relevant laboratory examination,karyotype,pelvic B ultrasound,pelvic magnetic resonance imaging(MRI)and AR gene mutation.The peripheral blood of the child and his parents were drawn,and peripheral blood DNA was extracted.The polymerase chain reaction(PCR)-DNA sequencing method was used to amplify all exons of the AR gene in the child and his parents.Then,they were directly sequenced.Results A 7-years and 2-months old child who suffered from DSD,revealed physical examination that the child had normal female external genitalia,as the clitoris length was 2.0 cm×0.8 cm,with visible vaginal opening,and there were masses at bilateral inguinal region,with a size of 1.5 cm×0.8 cm.The results of human chorionic gonadotropin(HCG)stimulation test:testosterone was 0 nmol/L,androstenedione was 1.78 nmol/L,dihydrotestosterone was 0.07 nmol/L before HCG was injected;but testosterone was 4.69 nmol/L,androstenedione was 2.10 nmol/L,dihydrotestosterone was 0.33 nmol/L after HCG was injection.Sex chromosome analysis reported 46,XY karyotype.Pelvic B ultrasound revealed the absence of a uterus and ovaries and the presence of bilateral testes like gonad at each side of internal inguinal ring,with a size of 1.4 cm×1.0 cm×0.8 cm in the left,1.5 cm×0.7 cm×0.8 cm in the right;but the kidney,ureter,urinary bladder,adrenal gland and retroperitoneal for B ultrasound revealed no abnormality.Pelvic MRI(non-enhanced and enhanced)showed the presence of a blind ending vagina between rectum and urinary bladder(40 mm in depth)and the absence of uterus and ovarian tissue.DNA sequencing found one c.1685T>C heterozygous mutation(p.Ile562Thr)on exon 2 of AR gene in the child.But retrieving and summarzing documents of the domestic and foreign information databases and websites,the locus mutation of AR gene had never been reported.The structure prediction of the mutated protein(Polyohen2 and SIFT software)was significantly changed.By verifying the locus site of the parents of this child,it was found that his mother carried the same mutation,but his father was found to be normal.Conclusions A c.1685 T>C mutation(p.Ile562Thr)on exon 2 of AR gene is a novel mutation.Combined with the patient's clinical manifestations and computer prediction results,it may suggest that the novel mutation of AR gene can lead to the occurrence of AIS.

Pseudorabies virus (PRV) is a swine herpesvirus of the Alphaherpesvirinae subfamily and a pathogen of swine resulting in devastating disease and economic losses worldwide. Cre/loxP site-specific system has the character of site specific, time specific, tissue specific and high efficiency in recombination, which makes this system universal in vivo and in vitro recombination of bacteria, fungus, plants, insects and mammals. A recombinant PRV which contain a loxP site in TK locus by using Cre/LoxP recombinant system was construsted. A pair of primers were synthesized according to the pEGFP-C1 sequence published on GenBank, and were used to amplify the EGFP gene expression cassette with two loxP sites flanking each side. This target gene was cloned into pSKLR, the resulting transfer vector pSKLR-GFP-loxP was then cotransfected into 293T cells with PRV SH strain genomic DNA. The recombinant virus rPRV1 was selected and purified in TK-143 cells by choosing fluorescent expressing plaques. Cre expression vector pOG231 was cotransfected into 293T cells with rPRV1 genomic DNA. The second recombinant virus rPRV2 was obtained, which contains only one loxP site in TK locus. Sequencing results of rPRV2 TK gene indicated that 34bp loxP site was inserted into rPRV2 genome and there were 270bp deletion in TK gene. PCR amplifying different generations of rPRV2 TK gene showed that the mutant was stable when passages in RK-13 cells. TCID_ 50 assay indicated that rPRV2 grows well on RK-13 cells. The LD_ 50 test results on BALB/C mice suggested that the virulence of rPRV2 was reduced. As a conclusion, the report gene GFP expression cassette was removed successfully from rPRV1 genome and only one LoxP site was leaved in rPRV2 genome by using Cre/LoxP recombinant system.

0.05).Conclusion Indexes of oxygen function may become criteria of early diagnosing NRDS,observing effect of treatment and guidance of ventilation weaning.

Objective To discuss the effect of clinical pathway in surgical new nurse' s training.Methods 80 new nurses were divided into two groups,traditional group and pathway group.Traditional group 40nurses adopted the traditional training methods,and pathway group 40 nurses according to clinical training path made training plans.Both groups were examined at the middle and the end of training.Restllts Acceptance rate in pathway group was 72.5％,and 92.5％ in traditional group,and the difference between the two groups was significant (x2 =5.541,P ＜0.05).Satisfaction rate in traditional group was 82.5％ and 97.5％ in pathway group,and the difference was also significant (x2 =4.114,P ＜ 0.05 ).Conclusions Application of clinical pathway training new nurses is a kind of systematization and standardization of the training mode.It not only can improve the satisfaction of new nurses in training,but also cultivate more excellent nurses for clinical.

Cancer, an abnormal cell proliferation resulted from multi-factors,has the highest morbidity and mortality among all the serious diseases. Considerable progress has been made in cancer biology in recent years. Tumor immunology, cancer stem cells (CSCs), autophagy, and epithelial-mesenchymal transition (EMT) have become hot topics of interests in this area. Detailed dissection of these biological processes will provide novel directions, targets, and strategies for the pharmacological evaluation, mechanism elucidation, and new drug development of traditional Chinese medicine.
Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Humans ; Neoplasms ; drug therapy ; genetics ; immunology ; physiopathology

AIM:To investigate the effects of 3-phosphoinositide-dependent protein-1(PDK1)on the biologi-cal characteristics of non-small-cell lung cancer cell line A549 and the underlying mechanisms.METHODS:The expres-sion levels of PDK1 in lung normal epithelial cell line BEAS-2B and different lung cancer cell lines H 460, SPCA1 and A549 were determined by Western blot and real-time PCR.Small interfering RNA was used to down-regulated PDK1 ex-pression in the A549 cells,and then cell viability and apoptosis were measured by CCK-8 assay and flow cytometry,respec-tively.The expression of cell cycle-and apoptosis-related molecules at protein level and the activation of Akt /FoxO1 path-way were measured by Western blot.Insulin-like growth factor-1(IGF-1,one of the most potent Akt activators)was used to evaluate the interaction between PDK 1 and Akt/FoxO1 pathway.RESULTS:Compared with lung normal epithelial cell line BEAS-2B,PDK1 expression in the lung cancer cell lines was obviously increased(P<0.05).Knockdown of PDK1 suppressed cell viability and cell cycle,but promoted the apoptosis of the A 549 cells.The results of Western blot showed that the protein levels of cyclin D1,CDK4,p-Rb,Bcl-2,p-Akt and cytoplasmic p-FoxO1 were significantly decreased after knockdown of PDK1,with increases in the protein levels of P27, cleaved caspase-3 and nuclear FoxO1.Pre-incubation with IGF-1 partly reversed the effect of PDK1 knockdown on Akt/FoxO1 pathway and increased the viability of A 549 cells. CONCLUSION:In human non-small-cell lung cancer A549 cells,knockdown of PDK1 suppresses cell viability and pro-motes cell apoptosis by regulating the expression of cell cycle-and apoptosis-related molecules via Akt/FoxO1 pathway, suggesting that PDK1 may be a potential target for diagnosis and theatment of lung cancer.

OBJECTIVETo compare the difference in the clinical efficacy on cervical spondylosis between acupuncture at three lines of cervical Jiaji (EX-B 2) and oral administration of jingfukang granules.

METHODSThree hundred cases of cervical spondylosis were divided into an acupuncture group and a medication group, 150 cases in each one. In the acupuncture group, according to the different types of cervical spondylosis, acupuncture was applied at three lines of cervical Jiaji (EX-B 2), once a day. In the medication group, jingfukang granules were prescribed for oral administration, one bag each time, three times a day. The treatment of ten days made one session in the two groups and two sessions were required totally. Before and after two sessions of treatment, the clinical assessment scale for cervical spondylosis (CASCS) was adopted to evaluate the score of subjective symptoms, clinical physical signs and adaptability as well as the total score in the patients of the two groups and the efficacy was compared.

RESULTSThe patients' symptoms and physical signs were alleviated, the adaptability was improved and the score of each item and the total score were increased in the two groups after treatment (all P<0.01). The improvements in the acupuncture group were better than those in the medication group (all P<0.01). The curative and markedly effective rate was 90.7% (136/150) in the acupuncture group, better than 66.0% (99/150) in the medication group (P<0.01).

CONCLUSIONAcupuncture at three lines of cervical Jiaji (EX-B 2) achieves the significant clinical efficacy on cervical spondylosis. This therapy is superior to relieving symptoms and physical signs and recovering adaptability as compared with jingfukang granules.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Female ; Humans ; Male ; Middle Aged ; Spondylosis ; therapy ; Treatment Outcome ; Young Adult

Objective To understand the status of compliance behaviors in adultescent with type 2 diabetes in community and to analyze the influence factors.Methods 288 adultescent with type 2 diabetes were investigated by using social demography material questionnaire,the obeying medical behavior meter,the society supports meter,the healthy faith meter through the group sampling.Results The score of obeying medical behavior was (56.8 ± 10.9 ),the score rate was 67.62％ ; drug therapy and dietary therapy were implemented best,followed by movement therapy,following-up in time,dealing with hypoglycemia,self-supervised and foot nursing were in the last.The total score of social support was (42.6 ±7.6),the score rate was 72.2％ ; The total score of healthy faith was ( 156.0 ± 16.4),the score rate was 73.58％.The social support was subjective support,objective support and support accessibility in the sequence of intensity; healthy faith was the obstacle sensation to the motion,the sensation to disease gravity,the sensation to the deference doctor's advice profit,moves the clue,the sensation to disease susceptivity,there was a positive correlation between compliance behaviors and social support( r =0.20,P ＜0.01 ),so did the compliance behaviors and healthy faith ( r =0.33,P ＜0.01 ),but no correlation between support accessibility and compliance behaviors (r =0.005,P ＞ 0.05 ).Except dealing with hypoglycemia,obstacle sensation had negative correlation with compliance behaviors,others had the positive correlation.Conclusions Compliance behaviors in adultescent with type 2 diabetes in community are uncommon,Healthy faith,subjective and objective social supports are factors which influence the compliance behavior.