Pathological features and survival of triple positive breast cancer in 271 cases / 中国现代普通外科进展

Objective:To explore pathological features and survival of triple positive breast cancer (TPBC).Methods:The clinical data of 271 cases of triple positive breast cancer from January 2010 to January 2017 in Suqian area were collected,compared with 283 cases of Luminal B I (HER2 negative).The clinical pathological features and survival were analyzed.Results:Among 271 cases of triple positive breast cancer,there were 89 cases (32.84％) of distant recurrence and metastasis in 2 years,and 137 cases (50.55％) of distant recurrence in 5 years.Among 283 cases of Luminal B I,there were 32 cases (11.31 ％) of distant recurrence and metastasis in 2 years.and 52 cases (18.37％) of distant recurrence in 5 years.There were significantly differences(P＜0.05).1 year Disease-free survival (DFS)and Overall survival (OS) of all patients were 100％,Among 271 cases of triple positive breast cancer,2-year DFS and OS were 64.94 ％,85.24％ respectively.3-year DFS and OS were 54.98 ％,69.74％ respectively,5-year DFS and OS were 43.54％,47.23％ respectively.Among 283 cases of Luminal B I,2-year DFS and OS were 86.22 ％,95.76％ respectively.3-year DFS and OS were 81.98 ％,80.92％ respectively,5-year DFS and OS were 76.33％,67.49％ respectively.There were significantly differences(P＜0.05).Conclusion:TPBC has the characteristics of poor biological behavior,large mass,pathological grade of grade Ⅲ,vascular or nerve infiltration,axillary lymph node metastasis,high proliferation index and high tumor load,and early distant recurrence,low DFS and OS.We Should choose individualized,targeted treatment programs,based on patient's hormone receptor and Ki67 expression,so as to benefit patients of TPBC.

Construction of VEGF gene-targeted small interfering RNA mediated by CMV promoter and its silencing effect / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To construct VEGF gene-targeted small interfering RNA (siRNA) and its expression vector driven by CMV promoter and to investigate its interference effect.</p><p><b>METHODS</b>The VEGF gene-targeted hairpin siRNA was designed, two complementary oligonucleotide strands were synthesized. After annealing, two-strand oligonucleotide was inserted into pDC311-SV40-RC vector, which was then identified by PCR and sequenced. Then human U-2 OS cell line was transfected with the vector using lipofectamine method. Finally, ELISA was performed to evaluate the expression of VEGF protein.</p><p><b>RESULTS</b>PCR-identification of positive clone and sequencing confirmed the vector containing the target siRNA. ELISA showed that compared with the control group, the expression levels of VEGF protein in transfected U-2 OS cells were decreased significantly (P<0.05).</p><p><b>CONCLUSION</b>VEGF gene-targeted siRNA and its vector mediated by CMV promoter were successfully constructed, which can reduce the VEGF protein expression after transfecting.</p>

Genetic polymorphism of dendritic cell-specific ICAM-3 grabbing nonintegrin and DC-SIGNR's exon 4 in Chinese hepatitis C patients / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study into the genetic polymorphism of DC-SIGN and DC-SIGNR's exon 4 in Chinese hepatitis C patients and its relationship with HCV infection susceptibility.</p><p><b>METHODS</b>Patients with hepatitis C (n=300, group A) and healthy subjects (n=520, group B) were genotyped and analysed for the repeat sequence of polymorphism of DC-SIGN and DC-SIGNR's exon 4 using PCR and DNA sequencing.</p><p><b>RESULTS</b>The distribution of genotypes and alleles in DC-SIGN's exon 4 in the two groups did not differ significantly (P > 0.05). The difference of allele frequency in DC-SIGNR's exon 4 between the two groups was also not significant (P > 0.05). However, 9/5 genotype distribution frequency of DC-SIGNR's exon 4 in patients with hepatitis C was significantly higher than that in the healthy subjects (P < 0.05).</p><p><b>CONCLUSION</b>There is no significant correlation between the genetic polymorphism of DC-SIGN's exon 4 and HCV infection susceptibility. 9/5 genotype distribution frequency of DC-SIGNR's exon 4 in patients with hepatitis C is significantly higher and may be associated with HCV infection susceptibility.</p>

Clinical study on YMDD mutation and pre-core C region and core promotor region mutation of the patients with chronic hepatitis B virus receiving lamivudine therapy / 中国基层医药

Objective To mvesugate the Tate of YMDD mutation accompamed with pre-core(region and core promotor region mutation and the clinical significance.Methods YMDD mutation and pre-core(at 1896 nu- cleotide)region and core promotor region(at 1762.1764 nucleotide)mutation were detected from the 122 patients with chronic hepatitis B virus after receiving lamivudine treatment above 6 months.Results 40 cases were tested for YMDI)mutations in 122 HBV patients with lamivudine treatment,and the positive rate of YMDD mutation was 32.8 %.After YMDD mutation,ALT,AST and HBV DNA of the patients significantly increased(P0.05).Conclusion The patients with YMDD mutation had higher rate of pre-core region(at 1896 nucleotide)and basal core promotor region(at 1762, 1764 nucleotide)mutation than those without YMDD mutation,but there was no correlation between the mutation and the deterioration of disease condition and the bad prognosis.

Relationship between Shenzhen HBV genotype and precore/core promoter mutation and antiviral effects / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To study the relationship between hepatitis B virus genotyping Shenzhen isolates and HBV precore/core promoter mutation and antiviral effects.</p><p><b>METHODS</b>The HBV genotyping of 165 patients with HBV was carried out with mAbs ELISA. HBV precore/core promoter mutation was detected with gene chip technology in 24 patients with CHB. The relationship between HBV genotyping and interferon, lamivudine effects was analyzed.</p><p><b>RESULTS</b>(1) Out of 165 cases, 106 (64.2%) of type B but 48 (29.1%) of type C were found. Type B accounted for 95.4% in group ASC, and type C for 64.7%in group LC (P<0.05). (2) Precore/core promoter mutation was found in 16 cases (10 of type B, and 6 of type C) out of 24 cases. Out of 16 cases, precore/core promoter mutation (nt1896, 1862) was found in 10 cases (9 cases of type B and 1 case of type C), while basal core promoter mutation (BCP mutation, nt1762,1764) was found in 6 cases (1 case of type B and 5 of type C). (3) Among 27 patients with CHB HBAg (+) treated with interferon, 11 cases of type B but 1 case of type C were tested to be fully responsive to interferon. Among 29 patients with CHB HBAg (+) treated with lamivudine, 15 cases of type B but 3 cases of type C were tested to be continuously responsive to lamivudine.</p><p><b>CONCLUSION</b>(1) HBV genotype popularity in Shenzhen area was classified as type B the first and type C the second. (2) Type C seems more apt to develop BCP mutation and cirrhosis, and to be less responsive to interferon or lamivudine.</p>

Molecular cloning and expression of the severe acute respiratory syndrome-associated coronavirus nucleocapsid protein and its clinical application / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To clone and express nucleocapsid (N) protein of the severe acute respiratory syndrome (SARS)-associated coronavirus, and to evaluate its antigenicity and application value in the development of serological diagnostic test for SARS.</p><p><b>METHODS</b>SARS-associated coronavirus N protein gene was amplified from its genomic RNA by reverse transcript nested polymerase chain reaction (RT-nested-PCR) and cloned into pBAD/Thio-TOPO prokaryotic expression vector. The recombinant N fusion protein was expressed and purified, and its antigenicity and specificity was analyzed by Western Blot, to establish the recombinant N protein-based ELISA for detection of IgG antibodies to SARS-associated coronavirus, and SARS-associated coronavirus lysates-based ELISA was compared parallelly.</p><p><b>RESULTS</b>The recombinant expression vector produced high level of the N fusion protein after induction, and that protein was purified successfully by affinity chromatography and displayed higher antigenicity and specificity as compared with whole virus lysates.</p><p><b>CONCLUSION</b>The recombinant SARS-associated coronavirus N protein possessed better antigenicity and specificity and could be employed to establish a new, sensitive, and specific ELISA for SARS diagnosis.</p>

A field trial for evaluating the safety of recombinant human interferon alpha-2b for nasal spray / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To evaluate the safety of recombinant human interferon alpha-2b for nasal spray for the prevention of SARS and other upper respiratory viral infections.</p><p><b>METHODS</b>Field epidemiologic evaluation was conducted, the design was randomized and had a synchronously parallel control group. In the study, the drugs were given for five days and all subjects were followed up for ten days.</p><p><b>RESULTS</b>During the period of using interferon, body temperature of the experimental group was normal compared to the control group. Experimental group had more influenza-like symptoms than the control group (P < 0.05), such as headache (4.83%-7.09%), dizziness (7.17%-11.63%), lassitude (8.55%-15.06%), muscular soreness (4.43%-7.09%), pharynx dryness (12.10%-17.85%), angina (6.25%-8.72%), abdominal pain (2.30%-5.50%) and diarrhea (2.45%-5.66%). Most of side effects reached their peak with in the first 3 days. Except for pharynx dryness, the incidences of all other side effects declined after completion of the use of the trial drug, and incidences of some symptoms in experimental group were lower than those of the control group. There were no significant differences in the symptoms of cough and expectoration between the experimental group and the control group. The incidence of exanthem in the control group was significantly higher than that in the experimental group. The side effect of bloody nasal mucus was not observed in experimental group, which had been reported by other authors in several volunteer studies.</p><p><b>CONCLUSION</b>Using recombinant human interferon alpha-2b for nasal spray could lead to some influenza-like symptoms, however, all those symptoms were mild , reversible, and relieved after completion of the use of the trial drug. No serious side effects were found during the period of following up. The authors conclude that the drug is safe.</p>

Cloning of ACE-2 gene encoding the functional receptor for the SARS coronavirus and its expression in eukaryotic cells / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>Angiotensin-converting enzyme 2 (ACE-2) has been identified as a functional receptor of severe acute respiratory syndrome coronavirus (SARS-CoV), so its gene was cloned and eukaryotic expressed for further insight into mechanisms in SARS-CoV entry and pathogenesis, as well as development of a safe and reliable neutralization assay for SARS-CoV.</p><p><b>METHODS</b>Total RNA was extracted from right atrial tissue of a patient with right heart failure resected during a valvular replacement surgery by Trizol one-step method, and the full-length ACE-2 encoding gene was acquired by RT-nested-PCR. The ACE-2 encoding gene was then cloned into pcDNA4/HisMax-TOPO eukaryotic expression vector to construct the recombinant plasmid pcDNA4/ ACE-2, which was then transfected into 293 T cell and ACE-2 eukaryotic transient expression was detected by Western Blot. Syncytia inhibition assay was established to detect SARS-CoV neutralizing antibody, and compared parallelly with SARS pseudovirus neutralization assay.</p><p><b>RESULTS</b>The recombinant plasmid pcDNA4/ ACE-2 could express ACE-2 protein in eukaryotic cells and induce cell-cell fusion between S protein- and ACE2-expressing cells. This cell-cell fusion assay could be used to detect SARS-CoV neutralizing antibody.</p><p><b>CONCLUSION</b>SARS-CoV receptor ACE-2 gene was successfully cloned and eukaryotic expressed, and used to establish syncytia inhibition assay for SARS-CoV neutralizing antibody assay.</p>

High-flux detection of HBV lamivudine resistant strains by genechip and analysis of mutant hotspots in those strains / 中华肝脏病杂志

<p><b>OBJECTIVE</b>Lamivudine resistant HBV strains in Shenzhen were detected at multiple sites and in large amounts to understand further the distribution of lamivudine resistant mutants.</p><p><b>METHODS</b>552 Hepatitis B patients's sera were examined using genechip method. Among them, 192 samples of lamivudine resistant mutant were further analyzed.</p><p><b>RESULTS</b>In those 192 lamivudine resistant samples, 191 were YMDD mutants, 124 mutants of codon 528 and 9 mutants of codon 555. 88% YMDD mutants were multi-mutants of YVDD and codon 528; single mutants of YIDD; multi-mutants of YIDD and codon 528. 91% codon of YMDD mutants were GTG, ATT; the other 9% were ATA, ATC.</p><p><b>CONCLUSIONS</b>These results suggest that mutants of codon 552 (YMDD) are core mutants. Mutants of codon 528 and 555 are incidental mutants, YVDD mutants always emerge with mutants of codon 528, but YIDD mutants appear differently. 9% YMDD mutants's codons are ATA or ATC. This may be the reason for the low positive rate shown by using the conventional PCR methods.</p>

Construction of small interfering RNA targeting heparanase gene and its inhibitory effect on the in-vasiveness of human malignant melanoma cell line A375 in vitro / 中华皮肤科杂志

Objective To construct the small interfering RNA (siRNA) targeting heparanase gene and its expressing vector,and to observe its interference effect on the expression of heparanase gene and inhibitory effect on the invasive potential of human malignant melanoma A375 cells.Methods Three siRNAs were designed.The recombinant plasmid pRNATU6.1/heparanase-siRNA was designed and constructed. A375 cells were cultured,and transfected with pRNATU6.1/heparanase-siRNA.The cells treated with lipo- fectamine or Opti-MEM served as the controls.Real-time fluorescence quantitative PCR and Western blot were performed to evaluate the expression of heparanase RNA and protein in these treated A375 cells.The in vitro invasive potential of treated A375 cells was assessed by Matrigel gel assay.Results The siRNA targeting heparanase gene was successfully cloned to the eukaryotic expressing vector pRNATU6.1.The expression levels of both heparanase RNA and protein decreased significantly in siRNA-transfected A375 cells than those in the control cells.The in vitro invasive potential of siRNA-transfected cells was also signifi- cantly inhibited as compared with that of the control cells (P

Expression of B7-CD28/CTLA-4 in peripheral blood lymphocytes of female patients with genital condyloma acuminatum / 中华皮肤科杂志

Objective To investigate the significance of B7-CD28/CTLA-4 (CD152),co-stimulatory molecules of T lymphocytes,in the onset,development and prognosis of genital congdyloma acuminatum (CA) in females.Methods Flow cytometry was utilized to detect the expression levels of CD80/CD86 in peripheral blood lymphocytes and of CD28/CD152 (CTLA-4) in CD4~+/CD8~+ T lymphocytes from 30 CA patients (17 primary CA,13 recurrent CA,15 at recovery stage of CA) and 15 healthy volunteers as con- trols.Results No significant difference was found for the frequencies of CD80~+,CD86~+,CD4~+/CD28~+ and CD8~+/CD28~+ lymphocytes between the primary or recurrent group and the control group.The frequencies of CD8~+/CD28~+,CD4~+/CD28~+ and CD80~+ lymphocytes were significantly higher in the recovery group than those in the recurrent group (P0.05).Conclusions There is an abnormal expression of co-stimulatory molecules B7-CD28/CTLA-4 (CD152) in periphoral blood lym- phocytes,CD4~+ and CD8~+ T lymphocytes in female patients with genital CA,and the expression abnormaility is closely linked with different disease stages of CA.

Early indexes to predict the therapeutic effect of interferon on chronic hepatitis B / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To summarize the clinical changing characters of the clinical markers after interferon treatment in chronic hepatitis B (CHB) and make out practical indexes to predict the effect.</p><p><b>METHODS</b>150 CHB patients were randomly divided into two groups: therapeutic group (90) and control group (60) in the prospective controlled trial. The levels of endogenous interferon before treatment, interferon antibody at the end of the second month and fourth month after treatment, alanine aminotransferase (ALT) and HBV DNA in the serum were detected. Then the data was analysed to find out indexes for predicting the effect.</p><p><b>RESULTS</b>(1) The clearance rate of HBeAg had no significant difference in age except for 20 - 30 and 30 - 40 (t > 2.331 2, P < 0.01). (2) It was more effective if ALT level was higher than 400 U/L before treatment and it decreased more than 50% two months after treatment. (3) The patients whose HBV DNA was negative (dot hybridization) or less than 10(6) copies/ml before treatment had higher rate of HBeAg clearance. (4) There was no effect on patients whose interferon antibody turned positive at the end of the second month. (5)A predictive method of comprehensive factors was made out, whose sensitivity, specificity, and accuracy were 80%, 100% and 90%, respectively.</p><p><b>CONCLUSION</b>The clinical characters of these Chinese patients are different from those of the westerners and the effects of interferon have close relation to the levels of ALT, HBV DNA and interferon antibody.</p>