[Progress and challenge of vaccine development against 2019-novel coronavirus (2019-nCoV)].

The outbreak of 2019-novel coronavirus (2019-nCoV) infection poses a serious threat to global public health. Vaccination is an effective way to prevent the epidemic of the virus. 2019-nCoV along with severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle East respiratory syndrome coronavirus (MERS-CoV) belong to the same ß-genus of coronavirus family. Basing on the previous experience and the technical platform of developing SARS-CoV and MERS-CoV vaccines, scientists from all over the world are working hard and quickly on the related fields. There are substantial progress in these fields including characterizing the 2019-nCoV virus, identification of candidate antigens and epitopes, establishment of animal models, characterizing the immune responses, and the design of vaccines. The development of 2019-nCoV vaccines covers all types: inactivated virus vaccine, recombinant protein vaccine, viral vector-based vaccine, mRNA vaccine, and DNA vaccine, et al. As of March 2020, two 2019-nCoV vaccines have entered phase I clinical trials. One is named as Ad5-nCoV developed by the Chinese Institute of Biotechnology of the Academy of Military Medical Sciences and Tianjin Cansino Biotechnology Inc. Ad5-nCoV is based on the replication-defective adenovirus type 5 as the vector to express 2019-nCoV spike protein. The another vaccine is mRNA-1273 developed by the National Institute of Allergy and Infectious Diseases and Moderna, Inc.. RNA-1273 is an mRNA vaccine expressing 2019-nCoV spike protein. Although the rapid development of 2019-nCoV vaccine, it still faces many unknown challenges, including the antigenic characteristics of the 2019-nCoV, the influence of antigenic variation, the protective immune response of host, the protection of the elderly population, and the downstream manufacturing process of the new vaccine. The safety and efficacy of vaccines are the first priority for vaccine development and should be carefully evaluated.

A lethal pneumonia model of Acinetobacter baumannii: an investigation in immunocompetent mice.

OBJECTIVES: Acinetobacter baumannii can cause severe nosocomial and community-acquired pneumonia. To study the pathogenesis of A. baumannii and to develop new treatments, appropriate mouse models are needed. Most reported mouse models of pulmonary A. baumannii infection are non-lethal or require mouse immunosuppression to enhance infection. These models are not suitable for studying host immune responses or evaluating immunotherapies. METHODS: The virulence of 30 clinical isolates was assessed in mice. The most virulent isolate, SJZ24, was selected to develop a pneumonia model in immunocompetent mice. The cytokine mRNA expression in the lung was assessed with real-time PCR. The cell infiltration in bronchoalveolar lavage fluid (BALF) after SJZ24 infection was determined by flow cytometry. Vaccine efficacy was assessed using this model. RESULTS: Intratracheal inoculation of SJZ24 (5 × 107 CFU) resulted in death in 100% of the mice (5/5). SJZ24-infected mice showed high bacterial burdens in blood and organs as well as severe lung-tissue damage. Infection with SJZ24 induced increased inflammatory cytokine expression in the lung and increased neutrophil infiltration in BALF. Immunization with inactivated whole cells of SJZ24 showed 100% protection (5/5) against A. baumanni infection in this model. CONCLUSIONS: We established a lethal pneumonia model in immunocompetent mice with hypervirulent A. baumannii isolate SJZ24. This model can be used to study the immune response to A. baumannii infection and to evaluate vaccine efficacy.

Controlled defect creation and removal in graphene and MoS2 monolayers.

It is known that defects strongly influence the properties of two-dimensional (2D) materials. The controlled creation and removal of defects can be utilized to tailor the optical and electronic responses of these 2D materials for optoelectronic and nanoelectronic applications. In this study, we developed an efficient approach to reversibly control the defect states in mechanically exfoliated graphene and molybdenum disulfide (MoS2) monolayers. The defects were created by aluminium oxide (Al2O3) plasmas and removed by moderate thermal annealing at up to 300 °C. We employed Raman and photoluminescence (PL) as well as electrical characterization to monitor the variation of the defect level in graphene and MoS2. For graphene, Raman spectra indicate that the Al2O3 plasma induced sp3-type defects with a controlled concentration, which have been substantially removed after thermal annealing. A similar trend was also observed in monolayer MoS2, as revealed by the defect-related emission peak (Xb) in the PL spectra. We further showed that the defects induced by the Al2O3 plasma in both 2D materials can be restored to any intended level via annealing under well-controlled conditions. Our work presents a new route to the functional design of the optical and electronic properties of graphene and MoS2-based devices through defect engineering.

MicroRNA-22 inhibits tumor growth and metastasis in gastric cancer by directly targeting MMP14 and Snail.

MicroRNAs (miRNAs) deregulation is frequent in human gastric cancers (GCs), but the role of specific miRNAs involved in this disease remains elusive. MiR-22 was previously reported to act as tumor suppressors or oncogenes in diverse cancers. However, their accurate expression, function and mechanism in GC are largely unclear. Here, we found that the expression of miR-22 was significantly reduced in clinical GC tissues compared with paired adjacent normal tissues, and was significantly correlated with a more aggressive phenotype of GC in patients, and miR-22 low expression correlated with poor overall survival. The introduction of miR-22 markedly suppressed GC cell growth, migration and invasion, and inhibition of miR-22 promoted GC cell proliferation, migration and invasion in vitro. We further demonstrated that miR-22 acted as tumor suppressors through targeting extracellular matrix (ECM) remodeling member matrix metalloproteinase 14 (MMP14) and epithelial-to-mesenchymal transition (EMT) inducer Snail in GC. Moreover, ectopic expression of MMP14 or Snail restored inhibitory effects of miR-22 on cell migration and invasion in GC cells, and a negative relationship between the miR-22 expression and MMP14 or Snail mRNA levels was observed in GC. Finally, overexpression of miR-22 suppressed tumor growth, peritoneal dissemination and pulmonary metastasis in vivo. Taken together, we identified that miR-22 is a potent tumor suppressor in GC. MiR-22 downregulation promotes GC invasion and metastasis by upregulating MMP14 and Snail, and then inducing ECM remodeling and EMT. These findings provide a better understanding of the development and progression of GC and may be an important implication for future therapy of the GC.

MicroRNA-141 inhibits tumor growth and metastasis in gastric cancer by directly targeting transcriptional co-activator with PDZ-binding motif, TAZ.

Gastric cancer (GC) is a biologically heterogeneous disease accompanying various genetic and epigenetic alterations, and the molecular mechanisms underlying this disease are complex and not completely understood. Increasing evidence shows that abnormal microRNA (miRNA) expression is involved in GC tumorigenesis, but the role of specific miRNAs involved in this disease remains elusive. MiR-141 was previously reported to act as tumor suppressors or oncogenes in diverse cancers. However, their accurate expression, function and mechanism in GC are largely unclear. Here we found that the expression of miR-141 was significantly reduced in GC compared with paired adjacent normal tissues and was significantly correlated with a more aggressive phenotype of GC in patients. Ectopic expression of miR-141 mimics in GC cell lines resulted in reduced proliferation, invasion and migration, and inhibition of miR-141 in GC cell lines promoted cell proliferation, invasion and migration in vitro. We further demonstrated that miR-141 acted as tumor suppressors through targeting transcriptional co-activator with PDZ-binding motif (TAZ) in GC. Moreover, the inverse relationship between miR-141 and its target was verified in patients and xenograft mice. Finally, overexpression of miR-141 suppressed tumor growth and pulmonary metastasis in nude mice. Take together, we identified that miR-141 is a potent tumor suppressor in the stomach, and its growth inhibitory effects are, in part, mediated through its downstream target gene, TAZ. These findings implied that miR-141 might be employed as novel prognostic markers and therapeutic targets of GC.

MicroRNA-25 promotes gastric cancer migration, invasion and proliferation by directly targeting transducer of ERBB2, 1 and correlates with poor survival.

Gastric cancer (GC) is one of the most common tumors and the molecular mechanism underlying its metastasis is still largely unclear. Here, we show that miR-25 was overexpressed in plasma and primary tumor tissues of GC patients with tumor node metastasis stage (III or IV) or lymph node metastasis. MiR-25 inhibition significantly decreased the metastasis, invasion and proliferation of GC cells in vitro, and reduced their capacity to develop distal pulmonary metastases and peritoneal dissemination in vivo. Furthermore, miR-25 repressed transducer of ERBB2, 1 (TOB1) expression by directly binding to TOB1-3'-UTR, and the inverse correlation was observed between the expressions of miR-25 and TOB1 mRNA in primary GC tissues. Moreover, the loss of TOB1 increased the metastasis, invasion and proliferation of GC cells, and the restoration of TOB1 led to suppressed metastasis, invasion and proliferation. The receiver operating characteristics analysis yielded an area under the curve value of 0.7325 in distinguishing the GC patients with death from those with survival. The analysis of optimal cutoff value revealed poor survival in GC patients with high plasma concentrations of miR-25 (>0.2333 amol/µl). Taken together, miR-25 promotes GC progression by directly downregulating TOB1 expression, and may be a noninvasive biomarker for the prognosis of GC patients.

High frequency variations of Helicobacter pylori isolates in individual hosts in a Chinese population.

BACKGROUND: Colonization of individual hosts by multiple Helicobacter pylori genotypes may be one reason why this infection is persistent and difficult to eradicate. METHODS: In order to study the diversity of H. pylori in individuals, a modified randomly amplified polymorphic DNA (RAPD) method was applied using primary culture isolates instead of passaged cultures. RESULTS: The results showed that variations in H. pylori were prevalent among individuals in the Chinese population, and the incidence of multiple colonization was 99.1% (115/116), significantly higher than in other reports. Moreover, the number of RAPD genotypes was found to be significantly associated with the process of disease development (p<0.05). Indeed, a trend for a higher number of RAPD genotypes within a single host (up to five genotypes) was observed as the disease developed or became more serious. After subculturing for three generations in our experiment, some genotypes present in the primary cultures were lost. The different genotypes in one patient may have originated from a single ancestral strain, as determined by analysis of six H. pylori housekeeping gene alleles, most of which were shown to be identical. CONCLUSIONS: These results suggest that investigating isolates of the primary culture will better reflect the H. pylori diversity in individuals. Also, they indicate that continuous variation of one strain in the gastric microenvironment may be the main cause of H. pylori diversity in individuals in the Chinese population.

Core genome haplotype diversity and vacA allelic heterogeneity of Chinese Helicobacter pylori strains.

The human gastric pathogen, Helicobacter pylori, has co-evolved with its host and established itself in the human stomach possibly millions of years ago. Therefore, the diversity of this bacterium is important in its clinical manifestations. Our aim has been to evaluate the genetic diversity of 40 H. pylori clinical isolates from four different parts of China. The methods of multi-locus sequence typing and vacA allele genotyping were used to assess their genetic diversity. To discriminate MLST, the vacA genotype method was used to identify strains. Patients from the northern, eastern, southern, and southwestern parts of China were recruited randomly from the cities of Beijing, Shanghai, Guangzhou, and Chongqing, respectively. Most of the sequence types are new and have never been reported in the database of the H. pylori multi-locus sequence typing system. The most prevalent vacA genotype in patients was s1a/m2 (80.0%), followed by s1b/m2 (17.5%). In contrast, the s1a/m1 genotype was scarcely represented (2.5%). The vacA genotype varied for each ST. These results showed that the MLST method offers high resolution of the H. pylori isolates in China when compared to vacA genotyping. The vacA allelic s1a has been correlated with the peptic ulcer. Because of the paucity of data on human isolates due to the absence of systematic investigations of H. pylori in China, the data provide useful information for understanding the epidemiology of H. pylori in China from the viewpoint of nucleotide sequence databases.

Immunological study of allogeneic mesenchymal stem cells during bone formation.

Autologous mesenchymal stem cells (MSCs) are limited in their clinical application because tissue-engineered bone cannot be pre-fabricated. Allogeneic MSCs are readily available but carry the risk of transplant rejection. It is not yet clear whether allogeneic MSCs can induce a rejection response during bone formation. In this study, two strains of genetically unmatched mini-pigs were used as experimental animals to study the immunological changes in MSCs in vitro and in vivo when generating bone. Mini-pig MSCs showed low immunogenicity during osteogenesis both in vitro and in vivo, indicating that allogeneic MSCs had little or no immunogenicity in osteosis. In conclusion, allogeneic MSCs are an important source of seed cells for the tissue engineering of bone. This favours the clinical application of pre-constructed tissue-engineered bone.

Interactions between organic flocculant PAN-DCD and dyes.

Organic flocculant polyacrylonitrile-dicyandiamide (PAN-DCD) was characterized by IR, 1H NMR and 13C NMR spectra. Polar groups, such as triazine, carboxyl, amidine, carbonamide and non-polar groups, such as methylene were identified in the macromolecular chain of PAN-DCD. The flocculant effectively decolorized the examined dyes in their aqueous solutions under acid condition. By comparing the IR spectra of dyes with that of PAN-DCD and that of flocs formed, weak chemical interactions between amino in PAN-DCD side chain and sulfonic acid group in the dye molecules were observed through the formation of -NH3+ SO3(-)-, NH2+ SO3(-)- or/and =NH+ SO3(-)-. By using equilibrium dialysis technique, the hydrophobic interactions between PAN-DCD and the dyes in the binding process were also observed from the experimental results. The binding extent of PAN-DCD to dye KN-R was greater than that of MO. The addition of urea into the binding system of KN-R by PAN-DCD reduced the first binding constant K from 12.2 x 10(5) (without urea) to 3.48 x 10(5) (with urea).

[Studies of the expression of human interleukin-2 receptor cDNA in mammalian cells].

Human interleukin-2 receptor cDNA was transfected into mouse fibroblast cells (L929) by using DNA-calcium phosphate coprecipitation method. Results from RNA dot-blot hybri dization, FITC-IL-2 fluorescent staining assay and anti-Tac specific rossette test demonstrated that the product of human interleukin-2 receptor cDNA in L929 cells is able to bind IL-2 and anti-Tac antibody. Moreover, the abnormal expression of IL-2R gene in T-cell leukemia cell lines such as Jukat cells and Molt-4 cells was analyzed and discussed.