Objective To study the expression of HPV、p16 and Her-2 in the squamous cell cervical carcinoma and its chnical significance. Methods Expression of HPV16/18、p16, and C-erbB-2 and the amplification of Her-2 gene were examined using situ hybridization technique,SP immunohistochemistry,and FISH imaging analysis system in 60 cases of cervical cancer, 61 cases of CIN, and 21 cases of normal cervical tissue,respectively.Results The positive rates of HPV16/18 and p16 in the normal tissue,CIN Ⅰ -Ⅱ,CINⅢ and the squamous cell cervical carcinoma were gradually increased, they wereo (0/21),9.68 ％ (3/31),46.67 ％ (14/30),71.67 ％ (43/60);0 (0/21),19.35 ％ (6/31),93.33 ％ (28/30),96.67 ％ (58/60),respectively,and there were significant differences among the groups (P＜0.05),but no significant difference was found between the normal tissue and the CIN Ⅰ - Ⅱ. The positive rates of Her-2 and Her-2 in the CIN Ⅲ and the carcinoma were 13.33 ％ (4/30),31.67 ％ (19/60),3.33 ％ (1/30),21.67 ％ (13/60),respectively,but in the normal group and the CIN Ⅰ - Ⅱ wereo,and the differences between the carcinoma group and the CIN group,the carcinoma group and the normal group were significant(P＜0.05).The expression of Her-2 and the amplification of Her-2 were closely related to the stage, degree of differentiation and metastasis of lymph node in the squamous cell cervical carcinoma (P＜0.05). Conclusion The infection of HPV is one of thetriggers for the squamous cell cervical carcinoma.The expression of p16 and the amplification of Her-2 may be closely correlated with tumor development and high expression of p16 and Her-2 indicates poor prognosis.

Objective To investigate the myocardial injury effects of intrathecal morphine injection in elderly patients undergoing thoracoscopic lobectomy.Methods Fifty-five elderly patients undergoing elective thoracoscopic lobectomy,28 males and 27 females,aged 65-85 years,BMI 18.5-27.9 kg/m2,ASA physical status Ⅰ or Ⅱ,were divided into two groups using the digital random allocation method:the control group(group C,n=28)and intrathecal morphine group(group M,n=27).Group M was given a single injection of morphine 4 pig/kg in the L2-3 space before surgery.General anesthesia was used in both groups,and single-lung ventilation was performed with double-lumen endotracheal intubation.Venous blood was collected before induction,24 and 48 hours after the operation to measure the levels of N-terminal brain natriuretic peptide precursor(NT-proBNP),creatine kinase isoenzyme(CK-MB),high-sensitivity troponin T(hs-TnT)and C-reactive protein(CRP).The incidence of myocardial injury after noncardiac surgery(MINS)was recorded.The intraoperative remifentanil dosage was recorded and the VAS pain scores at rest and during activity at 6,12,24,and 48 hours postoperatively were evaluated.The number of effective pa-tient-controlled intravenous analgesia(PCIA)compressions,the total number of PCIA compressions,the times of remedial analgesia in the postoperative period of 48 hours,as well as the incidence of postoperative adverse reactions(respiratory depression,nausea and vomiting,urinary retention,and pruritus)were re-corded.Postoperative 30-day major adverse cardiovascular and cerebrovascular events were recorded.Results Compared with preoperative,the levels of serum NT-proBNP,CK-MB,hs-TnT,and CRP were significantly higher in both groups at 24 and 48 hours postoperatively(P＜0.05).Compared with group C,the elevated levels of serum NT-proBNP,CK-MB,hs-TnT,CRP,and absolute hsTnT changes were signifi-cantly lower in group M 24 and 48 hours after operation(P＜0.05),the incidence of MINS was signifi-cantly lower in group M(P＜0.05).Compared with group C,the VAS pain scores of patients in group M were decreased significantly 6,12 and 24 hours during activity and 12 and 24 hours at rest after surgery(P＜0.05),the dosage of intraoperative remifentanil,the number of effective and total postoperative PCIA compressions,and the incidence of need for postoperative remedial analgesia were significantly reduced in group M(P＜0.05).There was no significantly difference in the incidence of postoperative adiverse reca-tions between the two groups.Conclusion Intrathecal morphine can reduce the levels of markers of myo-cardial injury in elderly patients undergoing thoracoscopic lobectomy,which plays a partial role in myocardial protection.

OBJECTIVETo study the severe acute respiratory syndrome (SARS)-associated coronavirus genotype and its characteristics.

METHODSA SARS-associated coronavirus isolate named ZJ01 was obtained from throat swab samples taken from a patient in Hangzhou, Zhejing province. The complete genome sequence of ZJ01 consisted of 29,715 bp (GenBank accession: AY297028, version: gi: 30910859). Seventeen SARS-associated coronavirus genome sequences in GenBank were compared to analyze the common sequence variations and the probability of co-occurrence of multiple polymorphisms or mutations. Phylogenetic analysis of those sequences was done.

RESULTSBy bioinformatics processing and analysis, the 5 loci nucleotides at ZJ01 genome were found being T, T, G, T and T, respectively. Compared with other SARS-associated coronavirus genomes in the GenBank database, an A/G mutation was detected besides the other 4 mutation loci (C:G:C:C/T:T:T:T) involved in this genetic signature. Therefore a new definition was put forward according to the 5 mutation loci. SARS-associated coronavirus strains would be grouped into two genotypes (C:G:A:C:C/T:T:G:T:T), and abbreviated as SARS coronavirus C genotype and T genotype. On the basis of this new definition, the ZJ01 isolate belongs to SARS-associated coronavirus T genotype, first discovered and reported in mainland China. Phylogenetic analysis of the spike protein gene fragments of these SARS-associated coronavirus strains showed that the GZ01 isolate was phylogenetically distinct from other isolates, and compared with groups F1 and F2 of the T genotype, the isolates of BJ01 and CUHK-W1 were more closely related to the GZ01 isolate. It was interesting to find that two (A/G and C/T) of the five mutation loci occurred in the spike protein gene, which caused changes of Asp to Gly and Thr to Ile in the protein, respectively.

CONCLUSIONAttention should be paid to whether these genotype and mutation patterns are related to the virus's biological activities,epidemic characteristics and host clinical symptoms.

Genotype ; Humans ; Middle Aged ; Mutation ; SARS Virus ; genetics

OBJECTIVETo investigate the effects of microRNA(miRNA)-29b on the proliferation and migration of breast cancer cells and its molecular mechanism.

METHODSThe recombinant lentiviral expression vector (lenti-miRNA-29b) was constructed and transfected into 293T cells to obtain lentivirus particles that were used to infect breast cancer MCF-7 cells. Transfection efficiency of lenti-miRNA-29b in MCF-7 cells was identified by the expression of green fluorescent protein (GFP). The expression of miRNA-29b was detected by real-time PCR. The cell proliferation and migration were detected by CCK8 assay and Transwell assay, respectively. The bioinformatics softwares were used to predict and screen the downstream target genes regulated by miRNA-29b, which were verified by double luciferase reporter gene assay, RT-PCR and Western blot. The effects of screened target gene RTKN on the growth and migration of MCF-7 cells were verified by RTKN siRNA.

RESULTSRecombinant lentiviral expression vector of miRNA-29b were successfully constructed. About 90% and 60% of the breast cancer cells showed green fluorescence in lenti-miRNA-29b and lenti-miRNA-NC groups, respectively. The expression of miRNA-29b in lenti-miRNA-29b group increased significantly compared with the lenti-miRNA-NC group and blank control group (all<0.05); the proliferation and migration ability of MCF-7 cells significantly reduced compared with the control group (all<0.05). The screening with bioinformatics softwares found that the 3'UTR coding region RTKN had the binding site to miRNA-29b; the dual luciferase reporter gene assay showed that the luciferase activity decreased significantly after the MCF-7 cells were co-transfected with wild type RTKN-WT-3'UTR and miRNA-29b mimics report gene vector (<0.05). The RTKN proteins in MCF-7 cells were significantly decreased after transfection with siRNA-RTKN, and the proliferation and migration ability of MCF-7 cells were significantly reduced (all<0.05).

CONCLUSIONSMiRNA-29b can inhibit the proliferation, invasion and metastasis of breast cancer cells by inhibiting the expression of RTKN.