To carry out phylogenetic analysis for drug-resistance genes from clinical isolates of Helicobacter pylori (Hp) among patients with gastric diseases from Tibet, China.

METHODSHp strains were isolated and cultured from saliva and gastric mucosal tissues derived from patients with gastric diseases. Nine strains (including 5 isolated from oral tissues, 1 isolated from gastric tissues, and 3 representative strains of SS international standard strains used for animal models) were tested for common antibiotic resistance. Together with an ACTT 11637 international standard strain, these were subjected to re-sequencing to obtain drug-resistance genes. Such genes from various sources were compared with the resistance genes of Hp strains recorded by the NCBI website. Combined with results of drug-resistance experiments, correlation between molecular evolution and drug-resistance was analyzed.

RESULTSTesting of gastric mucosal tissues and salivary samples from 217 patients has found 89 Hp strains, which yielded a total infection rate of 41.01%. The resistance rates of 9 representative Hp strains for clarithromycin, amoxicillin, metronidazole, levofloxacin and tetracycline were 77.8%, 77.8%, 44.4%, 77.8%, and 77.8%, respectively. Compared with the reference strain, the similarity between clarithromycin-resistance genes was 99%, and that between amoxicillin- and metronidazole-resistance genes was 96%-97%. A2143G mutation was also found in clarithromycin-resistant genes of three Hp strains.

CONCLUSIONThe sensitivity of Hp to metronidazole is much higher in patients from Tibet region, and the sensitivity of Hp to clarithromycin, amoxicillin, levofloxacin and tetracycline is poor. Resistance mutations are consistent with drug resistance.

Objective To analyze clinical characteristics of invasive Luminal subtype breast cancer.Methods The data of 162 invasive Luminal subtype breast cancer patients receiving operation in Cancer Hospital of Chinese Academy of Medical Science from January 1 st to September 30th in 2002,were collected and the clinical characteristics,recurrences,metastasis and survivals were retrospectively analyzed.Results The median time of follow-up was 92 months,ranging from 4 to 98 months.41 cases (25.3％,41/162) presented local recurrence or metastasis including 32 cases with metastasis ( 19.8％,32/162),2 cases with local recurrences (1.2％,2/162) and 7 cases with both local recurrence and metastasis (4.3％,7/162) ;Disease-free survival (DFS) and the 5-year DFS were 73.1％ and 79.6％,respectively.27 patients ( 16.7％,27/162) died of breast cancer,the overall survival (OS) and 5-year OS were 82.5％ and 85.3％,respectively.According to Kaplen-Meier survival analysis,tumor size,lymph node status and clinical stage were correlated to overall survival time ( P ＜ 0.05 ) ; and rumor size,lymph node starus,grade,clinical stage and PR status were correlated to disease-free survival time ( P ＜ 0.05 ).By multivariate analysis,TNM stage,PR and PCNA were independent prognostic factors correlated to overall survival time (OR=0.633,95％ CI:0.411 -0.976,P＜0.05; OR =0.823,95％ CI:1.012-3.283,P ＜ 0.05) ; TNM stage and PR was independent prognostic factors correlated to disease-free survival time (OR =3.273,95％ CI:1.719 - 6.232,P ＜ 0.01 ; OR =0.599,95％ CI:0.423 - 0.850,P ＜ 0.01 ).Conclusions In invasive Luminal subtype breast cancers,PR is correlated to fine prognosis,and PCNA is correlated to overall survival time.

Objective To explore the application value of neoadjuvant chemotherapy (NACT) in treating cervical carcinoma. Methods Seventy-one cervical carcinoma patients who were divided into three groups:PF group (cisplatin and fluorouracil,23 cases) or CBP group (carboplatin,bleomycetin and cyclophosphamide, 24 cases) or TP group (paclitaxel and cisplatin, 24 cases); operation was made 14 - 21 days afterwards. The therapeutic effect, chemotherapy side-effect and the effect on operation and pathology for these three groups were analyzed and compared. Results The effective rate was 93.8%(30/32),88.6% (31/35),82.4%(28/34) in TP group,CBP group,PF group,respectively. However, the therapeutic effect had no statistics significance with age, preoperative tumor grade, pathologic type for all the three groups, the therapeutic effect had statistical significance with clinical stage. The rate which pathological examination showed no residual cancer was biggest in CBP group [CBP group was 28.6%(6/21), TP group was 4.3% (1/23), PF group was 11.8%(2/17), P < 0.05 ]. Conclusions Three NACT projects are safe and effective treatment for cervical cancer. However,each project has advantages. The NACT projects can be elected for different patient according to his illness and economy.

Objective To evaluate the accuracy of core needle biopsy (CNB) in diagnosing breast masses and its coherence with immunohistochemical (IHC) examination results of estrogen receptor (ER), progesterone receptor (PR) and Her2 protein between pre-and post-chemotherapy in invasive breast cancer. Methods The results of 516 CNB cases from June, 2005 to April, 2008 were analyzed retrospectively. The pathological examination was performed by two pathologists independently. Results 484 cases of malignant tumor, carcinoma in situ and phyllodes tumor were found in this group with the sensitivity of 96.7%. Sixteen cases of false negative (3. 3% ) were demonstrated by surgical biopsy. The accurate rate of CNB was not influenced by the maximum diameter of masses ( P = O. 423 ). The agreement rate of IHC results of ER, PgR and Her2 between pre- and post-chemotherapy were 90. 3%, 76. 8% and 82.5%, respectively. Conclusion CNB is a useful diagnostic method with a satisfactory accuracy in any size of breast masses. Given the histological heterogeneity of invasive breast cancer and the influence of ehemotherapy, the coherence of prechemotherapy IHC for ER, PgR and Her2 is not optimal with that of post-chemotherapy.

In order to explore the feasibility and protective efficiency of influenza DNA vaccine, we constructed eukaryotic expressing plasmids encoding HA and HA1 of influenza A virus (A/PR/8/34) and studied their expression in HEK293 cells. HA and HA1 genes were amplified by RT-PCR and cloned into pcDNA3.1(+) to generate pcDNA3.1(+)/HA and pcDNA3.1(+)/HA1, respectively. After verification of the cloning fidelity by restriction endonuclease digestion, PCR, and sequencing, pcDNA3.1(+)/HA and pcDNA3.1(+)/HA1 were transfected into HEK293 cells using PolyFect Transfection Reagent. Immunofluorescence assay was used to detect the transient expressing cells. Fluorescence microscopy revealed strong expression of target gene in HEK293 cells transiently transfected with either pcDNA3.1(+)/HA or pcDNA3.1(+)/HA1. Therefore, the results confirm the successful construction of eukaryotic expressing plasmids capable of driving the eukaryotic expression of influenza virus antigen HA and HA1, which is likely to provide a basis for both further investigation of the mechanism of influenza viral infection and the development of influenza DNA vaccine.

In order to explore the feasibility and protective efficiency of influenza DNA vaccine, we constructed eukaryotic expressing plasmids encoding HA and HA1 of influenza A virus (A/PR/8/34) and studied their expression in HEK293 cells. HA and HA1 genes were amplified by RT-PCR and cloned into pcDNA3. 1 (+) to generate pcDNA3. 1 (+)/HA and pcDNA3.1 (+)/HA1, respectively. After verification of the cloning fidelity by restriction endonuclease digestion, PCR, and sequencing, pcDNA3.1 (+)/HA and pcDNA3.1 (+)/HA1 were transfected into HEK293 cells using PolyFect Transfection Reagent. Immunofluorescence assay was used to detect the transient expressing cells. Fluorescence microscopy revealed strong expression of target gene in HEK293 cells transiently transfected with either pcDNA3. 1 (+)/HA or pcDNA3. 1 (+)/HA1. Therefore, the results confirm the successful construction of eukaryotic expressing plasmids capable of driving the eukaryotic expression of influenza virus antigen HA and HA1, which is likely to provide a basis for both further investigation of the mechanism of influenza viral infection and the development of influenza DNA vaccine.

Objective To evaluate the localized biopsy of nonpalpable breast lesions (NPBLs) and its role in the early diagnosis and treatment of breast cancer. Methods One hundred and fifty-eight NPBLs from a series of 141 women detected by mammography were resected with wire localization technique. Results Forty-two lesions (26.6%, 42/158) in 42 patients were diagnosed with malignant result, including 12(28.6%) patients with stage 0 breast cancer, 24(57.1%) with stageⅠ, 2(4.8%) with stage Ⅱ and 4(9.5%) with stage Ⅲ disease according to American Joint Committee on Cancer (AJCC) staging system(the 6th edition). The contralateral axillary lymph nodes metastasis were found in only one (2.4%) patient with stage Ⅲ disease and the other forty-one patients remained free of recurrent disease at a median follow-up of 31 months.Conclusion The results showed that the most nonpalpable breast cancers detected by mammography were early-stage breast cancers and had good prognosis. The NPBLs should get a localized biopsy in order to facilitate the early diagnosis and treatment of nonpalpable breast cancers.

Hybrid linear analysis (HLA) was applied to resolution of overlapping spectra of Fe3+ -salicylfluorone and Al3+ -salicylfluorone complexes and simultaneous spectrophotometric determination of Fe3 + and Al3 + . The absorbance matrix of 7 standard mixtures at 41 measuring points ranged from the wavelength of 550 nm to 630 nm was used for calibration. To avoid the effect of interaction between the two components on the determination, the column vector of K matrix obtained from the standard mixtures with least squares was used as the pure spectrum of component. The recoveries of the two elements for the analysis of the synthetic samples were 93.3% ～ 107.5% in the range of the concentration ratio of Fe3+:Al3+ = 10:1 to 1:8. Comparing with the partial least squares (PLS) model,the HLA method was simple,accuracy and precise.

Objective To evaluate breast imaging reporting and data system (BI-RADS) in categorization and biopsy of nonpalpable breast lesions (NPBLs). MethodsOne hundred and sixty-two NPBLs from 144 female patients detected by mammography were retrospectively categorized according to BI-RADS. All the lesions finally got histopathological diagnosis by wire-localization biopsy. Results There were 46 malignant lesions, with the positive predictive value for cancer of 28.4% (46/162). The cases of BI-RADS 2-5 lesions were 11, 55, 77, and 19 after categorization according to the BI-RADS, and the positive predictive value of each category for cancer was 0% (0/11),3.6% (2/55),37.7% (29/77),78.9% (15/19) respectively. ConclusionBI-RADS categorization improves the diagnostic specificity of nonpalpable breast lesions and helps decision-making for biopsy. It is suggested that NPBLs on category of BI-RADS 4 or 5 should undergo biopsy.

Objective To evaluate the feasibility of sentinel lymph node biopsy(SLNB)during surgery of breast cancer. MethodsRadioactive colloid and blue dye were injected intradermally around the tumor seperately before the operation and the SLN were detected first by lymph scintigraphy. SLN was detected and located using ?-finder and the blue dye. Axillary lymph node dissection(ALND)was performed routinely after the SLNB. Results Among 116 breast cancer patients,this procedure was successful in 98.3% of cases. The sensitivity, accuracy and false negative rate were 93.6%, 97.4% and 6.4%, respectively. Conclusions SLNB is a simple, safe and reliable technique.Routine ALND could be raplaced by SLNB in breast cancer patients undergoing surgery.