Helicobacter pylori is a bacterium that can cause chronic gastritis, peptic ulcers, and other gastrointestinal diseases. The World Health Organization has classified H. pylori as a group Ⅰ carcinogen. Antibiotics are the primary clinical approach for eradicating H. pylori. However, incomplete eradication of H. pylori by antibiotics can lead to persistent infection, which is a major risk factor for the high incidence of gastric cancer. The widespread use of antibiotics has led to the emergence of multidrug resistance in H. pylori, contributing to treatment failures of chronic gastric diseases and increasing the risk of spreading resistant strains. Multidrug-resistant H. pylori has become a serious challenge in the diagnosis and treatment of gastrointestinal diseases. This paper reviews the global trends in the development of multidrug resistance in H. pylori, the underlying mechanisms, the challenges it poses to clinical diagnosis, and its impact on drug development, drawing on relevant literature and the research findings from our group. It proposes using cgt expression as a novel method for determining viable bacteria, identifying intracellularization as a new form of resistance in H. pylori, and exploring the potential of O-glycans as a therapeutic approach against H. pylori to address multidrug resistance. It provides new insights into understanding the mechanisms of H. pylori multidrug resistance and its prevention strategies, offering promising directions for future clinical treatments and antimicrobial drug development.
Helicobacter pylori/genetics* ; Humans ; Drug Resistance, Multiple, Bacterial ; Helicobacter Infections/microbiology* ; Anti-Bacterial Agents/therapeutic use* ; Gastrointestinal Diseases/drug therapy*

Objective To establish a viable bacteria assay for Helicobacter pylori(H.pylori)by assessing the cgt gene expression,and to develop accordingly a rapid and novel testing method for clinical precision treatment.Methods Viable bacteria count was determined in bacterial cultures.The transcriptional expression level of cgt(hp0421),the conserved gene that encodes cholesterol-α-glucosyltransferase(CGT)in H.pylori,was measured by RT-PCR.The correlation between the number of colonies and cgt gene transcription expression was analyzed and the regression model was constructed.The linear range,sensitivity,and specificity of the new method were examined accordingly.The bactericidal action of clarithromycin was assessed using this method to verify the performance of the method in determining clinical bacterial drug resistance.Results The Ct values of cgt for H.pylori colony counts of 102,104,106,and 108 CFU/mL were 29.67±0.14,23.37±0.36,17.65±0.37,and 11.38±0.39,respectively.In the range of 101-108 CFU/mL,the regression equation for cgt gene expression and viable bacterial counts determined by RT-qPCR was y=-0.350 1x+12.49,with the correlation coefficient being R2=0.9992 and the sensitivity being 101 CFU/mL,showing no cross-reaction with 13 other bacteria.The lg values of live H.pylori bacteria treated with clarithromycin at 0,5,10,20,and 40 μg/mL for 12 h were 2.57±0.02,2.45±0.01,2.19±0.02,1.91±0.07,and 1.33±0.05,respectively.The corresponding cgt gene Ct values were 27.76±0.09,28.37±0.24,29.51±0.14,30.11±0.12,and 31.66±0.11.By applying the cgt gene expression in the equation,the estimated counts of viable bacteria were found to be 2.73±0.03,2.52±0.08,2.11±0.05,1.89±0.02,and 1.33±0.04,showing no significant difference in statistical analysis(P＞0.05).Conclusion The method for assessing viable bacteria account by evaluating cgt gene expression in H.pylori was successfully established,significantly reducing the time required to determine viable bacteria count and providing a new method for clinical viable bacteria testing.

Helicobacter pylori(H.pylori),for a long time,has generally been considered an extracellular bacterium.However,recent findings have shown that H.pylori can gain entry into host cells,evade attacks from the host immune system and the killing ability of medication,form stable intracellular ecological niche,and achieve re-release into the extracellular environment,thus causing recurrent infections.H.pylori intracellular infection causes cellular signaling and metabolic alterations,which may be closely associated with the pathogenesis and progression of tumors,thereby presenting new challenges for clinical eradicative treatment of H.pylori.Herein,examining this issue from a clinical perspective,we reviewed reported findings on the mechanisms of how H.pylori achieved intracellular infection,including the breaching of the host cell biological barrier,immune evasion,and resistance to autophagy.In addition,we discussed our reflections and the prospects of important questions concerning H.pylori,including the clinical prevention and control strategy,intracellular derivation,and the damage to host cells.

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.