[The application of next-generation sequencing in the diagnosis of Pneumocystis pneumonia in non-HIV infected patients].

Objective: To evaluate the clinical value of next-generation sequencing in the diagnosis of Pneumocystis pneumonia in non-HIV infected patients. Methods: A retrospective study was conducted on the diagnosis and treatment of Pneumocystis pneumonia in 5 non-HIV patients in the Fourth Medical Center of the General Hospital of the PLA from September 1, 2017 to September 1, 2018. Next-generation sequencing of BALF were compared with the traditional laboratory microbiological test, and the advantages of the next-generation sequencing in the diagnosis of Pneumocystis pneumonia in non-HIV infected patients were analyzed. Results: There were 3 males and 2 females, with a mean age (48±6) years. Three patients had membranous nephropathy, a patient had tuberculous meningitis, and a patient had esophageal cancer after radiotherapy and chemotherapy. All patients had glucocorticoid medication history before. The clinical manifestations were fever, cough and dyspnea. The chest CT mainly showed bilateral lung ground glass shadows. All the results of 1, 3-ß-D-glucan test were more than 1 000 ng/L. Bronchoalveolar lavage was performed in the 5 cases, and Pneumocystis cysts were found in 1 BALF by Gomori's methenamine silver nitrate staining, and the DNAs of Pneumocystis and human herpesvirus were detected in 5 BALFs by next-generation sequencing. All patients were treated with sulfamethoxazole/trimethoprim (orally, 1.44 g, q8 h) for 23 to 72 days (median 33 days), and with ganciclovir(Ⅳ, 250 mg q12 h) for 6 to 22 days (median 15 days). The chest CT manifestations and symptoms were improved after treatment, without death. Conclusions: The next-generation sequencing of BALF is more specific and sensitive in the diagnosis of Pneumocystis pneumoniae in non-HIV patients. It is faster, more comprehensive and more accurate than the traditional laboratory test, and could be widely used as a PCP diagnosis technique.

Downregulated long non-coding RNA TRPM2-AS inhibits cisplatin resistance of non-small cell lung cancer cells via activation of p53- p66shc pathway.

OBJECTIVE: Non-small cell lung cancer (NSCLC), as an ordinary malignant tumor, presents with high death rate and poor prognosis. Few literatures have explored the association between NSCLC development and lncRNAs expression. This study focuses on the important role of a novel lncRNA TRPM2-AS in the development of chemo-resistance in NSCLC. MATERIALS AND METHODS: The expression level of lncRNA TRPM2-AS was identified by using qRT-PCR assay. The apoptosis rate and the alteration of the cell cycle were detected by the flow cytometric analysis. Cell Counting Kit-8 assay (CCK8) was utilized for detecting chemo-sensitivity of the cisplatin-resistant A549/DDP cells. The p53 and p66shc protein levels were detected by Western blotting assay. RESULTS: A549/DDP cells presented remarkably higher expression of lncRNA TRPM2-AS than paired A549 cells. Moreover, re-sensitization to cisplatin was seen in A549/DDP cells after lncRNA TRPM2-AS knockdown. On the contrary, the sensitivity of lncRNA TRPM2-AS-overexpressed A549 cells to cisplatin decreased obviously when compared with the control. Furthermore, downregulated lncRNA TRPM2-AS induced cell apoptosis and altered cell cycle distribution through activating the p53-p66shc pathway. CONCLUSIONS: We suggest that lncRNA TRPM2-AS participates in the resistance of NSCLC cells to cisplatin, which may provide a new therapeutic target of NSCLC.

Biological functions of recombinant human interleukin-2.

Recombinant interleukin-2 (rIL-2) is able to maintain the growth and multiplication of T-lymphocytes and IL-2 dependent CTLL-2 cells up to 28 days with the multiplication of T-lymphocytes as high as about 1000 fold. When rIL-2 and its monoclonal antibody were added together, the above functions of rIL-2 were specially blocked, showing that these biological functions were exerted specifically by rIL-2. The rIL-2 is also able to increase the natural killer (NK) activity up to 46-96%, while higher concentrations of rIL-2 might exhibit inhibitory effect. The cytotoxicity of lymphokine activated killer (LAK) cells induced by rIL-2 for killing HL-60 cell line and solid tumour (human lung carcinoma) reached 54.84% and 37.96%, respectively. All these results indicate that the biological functions of rIL-2 were quite similar to that of the natural one.

Study on the mechanism of interferon action (XI)--Effect of pppA2'p5'A on the level of cAMP and cGMP in macrophages.

In this paper, the increase of cellular cAMP and cGMP levels in macrophages induced by ppA2'p5' A2'p5'A (briefly 2'-5'P3A3) is first reported. The optimal concentration of 2'-5' P3A3 for the elevation of cellular cGMP to the highest level is 10(-7)-10(-6) mol/L, while that for cAMP is 10(-7) mol/L. The time for cGMP to reach its peak value is 15 min and that for cAMP is 2 h, when the cells are treated with 2'-5' P3A3 at 10(-7) mol/L, which is the optimal concentration for developing biological effect of macrophages (phagocytosis). These results suggest that cGMP and cAMP may be related to, or may be the mediators for, 2'-5'P3A3 action.

Mechanism of interferon action. Activation of macrophages and antagonism to alpha-feto-protein by pppA2'p5'A2'p5'A.

pppA2'p5'A2'p5'A(2'-5'P3A3) activates macrophages and increases the phagocytosis of macrophages from different species including human beings. This indicates that the activation of macrophages may be a general action of 2'-5'P3A3. This discovery broadens the effect of 2'-5'P3A3 beyond the antiviral field. alpha-Feto-protein (AFP) inhibits the phagocytosis of macrophages and may be involved in the development of hepatoma. Data presented here show that 2'-5'P3A3 can antagonize this suppressive effect of AFP. Methods used so far for introducing 2'-5'P3A3 into the cells were made with the aid of CaCl2, etc. under conditions which may not be the same as those used clinically. It was found that 2'-5'P3A3 can develop its biological effect without the aid of CaCl2, etc.