Objective The progression of pulmonary fibrosis is a common clinical issue after acute lung injury(ALI).We aimed to construct a model simulating clinical ALI-induced pulmonary fibrosis by repeated challenges with lipopolysaccharide(LPS).We then observed the development from ALI to pulmonary fibrosis,to explore the possible mechanisms mediating the transition from inflammatory injury to fibrosis.Methods Mice were treated with LPS(1,2,4,8 mg/kg)intranasally to induce ALI.At 7 d,14 d,21 d,28 d,35 d,and 42 d after modeling respectively,α-smooth muscle actin(SMA),collagen 1(Col-1),and hydroxyproline levels in lung tissue,and collagen fiber deposition were observed by Masson staining and compared to determine the process and degree of fibrosis formation in different modeling method.Expression changes in interleukin(IL)-1β,tumor necrosis factor(TNF)-α,and transforming growth factor(TGF)-β1 in lung tissue at each time point were detected to explore the mechanisms of fibrosis formation.Results Treatment of mice with 1,4,and 4 mg/kg LPS for 3 consecutive days(M-1 group)result ed in a stable ALI-induced pulmonary fibrosis model.Masson staining showed that α-SMA,Col-1,hydroxyproline,and collagen fiber deposition in the lung tissue began to increase in M-1 group mice in a time-dependent manner after 7 d.Collagen deposition in the lung tissue interstitium was significantly increased at 21 d post-modeling,and fibrosis indicators were significantly increased at 28 d,compared with control mice.Collagen deposition continued to increase until 42 d.Hydroxyproline and collagen fibers in the lung tissue in the other model groups with different doses and hit times did not increase significantly compared with the control group.TGF-β1 expression detected by Western blot began to increase gradually 14 d after modelling in the M-1 group,and was significantly higher than in the control group at 28 d.The pro-inflammatory cytokines TNF-α and IL-1β increased significantly on day 7(acute phase),and TNF-α expression continued to increase until 28 d,while IL-1β gradually decreased after day 7.TNF-α and IL-1β in the lung tissue both continued to decrease after the acute phase in the other model groups without fibrosis.Conclusions LPS 1,4,and 4 mg/kg for 3 consecutive days can be used to construct an ALI/acute respiratory distress syndrome-induced pulmonary fibrosis model,via a mechanism that may be related to the sustained high expression of TNF-α regulating TGF-β1 to induce fibroblast activation and proliferation.

Objective The progression of pulmonary fibrosis is a common clinical issue after acute lung injury(ALI).We aimed to construct a model simulating clinical ALI-induced pulmonary fibrosis by repeated challenges with lipopolysaccharide(LPS).We then observed the development from ALI to pulmonary fibrosis,to explore the possible mechanisms mediating the transition from inflammatory injury to fibrosis.Methods Mice were treated with LPS(1,2,4,8 mg/kg)intranasally to induce ALI.At 7 d,14 d,21 d,28 d,35 d,and 42 d after modeling respectively,α-smooth muscle actin(SMA),collagen 1(Col-1),and hydroxyproline levels in lung tissue,and collagen fiber deposition were observed by Masson staining and compared to determine the process and degree of fibrosis formation in different modeling method.Expression changes in interleukin(IL)-1β,tumor necrosis factor(TNF)-α,and transforming growth factor(TGF)-β1 in lung tissue at each time point were detected to explore the mechanisms of fibrosis formation.Results Treatment of mice with 1,4,and 4 mg/kg LPS for 3 consecutive days(M-1 group)result ed in a stable ALI-induced pulmonary fibrosis model.Masson staining showed that α-SMA,Col-1,hydroxyproline,and collagen fiber deposition in the lung tissue began to increase in M-1 group mice in a time-dependent manner after 7 d.Collagen deposition in the lung tissue interstitium was significantly increased at 21 d post-modeling,and fibrosis indicators were significantly increased at 28 d,compared with control mice.Collagen deposition continued to increase until 42 d.Hydroxyproline and collagen fibers in the lung tissue in the other model groups with different doses and hit times did not increase significantly compared with the control group.TGF-β1 expression detected by Western blot began to increase gradually 14 d after modelling in the M-1 group,and was significantly higher than in the control group at 28 d.The pro-inflammatory cytokines TNF-α and IL-1β increased significantly on day 7(acute phase),and TNF-α expression continued to increase until 28 d,while IL-1β gradually decreased after day 7.TNF-α and IL-1β in the lung tissue both continued to decrease after the acute phase in the other model groups without fibrosis.Conclusions LPS 1,4,and 4 mg/kg for 3 consecutive days can be used to construct an ALI/acute respiratory distress syndrome-induced pulmonary fibrosis model,via a mechanism that may be related to the sustained high expression of TNF-α regulating TGF-β1 to induce fibroblast activation and proliferation.

OBJECTIVE: To investigate the clinical value of oligoclonal bands (OB) in patients with multiple myeloma (MM). METHODS: The laboratory test and clinical data of 624 newly diagnosed MM patients admitted to Blood Diseases Hospital of Chinese Academy of Medical Sciences from January 2013 to December 2019 were retrospectively analyzed, including 30 patients with OB, and the clinical characteristics, treatment effects and survival of OB and non-OB patients were analyzed and compared. RESULTS: OB occurred in 11.8% (22/187) of patients who received autologous stem cell transplantation(ASCT) and only 1.8% (8/437) of patients who did not receive ASCT (P=0.000). The median time to the appearance of oligoclonal bands was 3.2(0.6-10.5) months after transplantation. The M protein types of oligoclonal bands mainly include IgG κ, IgG λ, IgM λ and λ light chains. In the presence of oligoclonal bands, 90% of patients were evaluated as complete remission (CR) and above. There were no statistically significant differences in disease stage, tumor burden, and genetic abnormalities between OB and non-OB patients. Among the all patients, the prognosis of OB patients was significantly better than that of non-OB patients, and OB patients showed deeper disease remission (significantly higher CR rate, MRD negative rate, and longer MRD negative duration). Among patients who underwent ASCT, OB patients showed earlier immune recovery, but the depth of treatment response and survival outcomes were similar between OB and non-OB patients, it was no statistically difference. Although OB patients showed earlier immune reconstitution, this did not translate into better survival, suggesting that the better prognosis of OB patients was mainly related to deeper and durable remission rather than early immune reconstitution. Further analysis in patients who received ASCT and obtained MRD negative indicated that there was no additional survival benefit in patients with OB. CONCLUSION The better prognosis of OB patients may be related to the deeper treatment response, but not to the early immune reconstitution. The appearance of OB is only a sign of deep remission and early immune reconstitution in patients, it cannot be translated into survival benefit of MM patients.
Hematopoietic Stem Cell Transplantation ; Humans ; Immunoglobulin G ; Immunoglobulin M ; Multiple Myeloma ; Oligoclonal Bands ; Retrospective Studies ; Transplantation, Autologous

OBJECTIVE: To investigate the comparability of the Freelite, Binding Site, Beckman and N Latex FLC, Siemens in the detection of serum free light chain (sFLC) . METHODS: Fifty newly diagnosed multiple myeloma (MM) patients in Tianjin Institute of Blood Research from November 2019 to February 2020 were enrolled. The two systems (Freelite, Binding Site, Beckman and N Latex FLC, Siemens) were used to detect the sFLC of the samples. Outlier detection was performed by ESD method, methodological comparison and deviation assessment were performed by Passing-Bablok regression and Bland-Altman regression. RESULTS: Both the systems could quantitatively analyze free kappa light chain serum samples and free lambda light chain samples. Freelite, Binding Site, Beckman and N Latex FLC, Siemens free light chain test showed FLC-κ：36.5 (6.5, 194), 40.5 (6.94, 288), FLC-λ: 30.1 (4.3, 170.5), 35.1 (2.28, 526), rFLC (FLC-κ/ FLC-λ) : 0.82 (0.05, 43.25), 1.03 (0.03, 32.04), dFLC (｜FLC-κ- FLC-λ｜) : -5.8 (-161.97, 183.7), 1.1 (-505.1, 279.01), which existed no outliers. There were systematic differences, and the deviation level was not within the clinically acceptable range. CONCLUSION Both the systems can meet the needs of clinical diagnosis and treatment, but there is a significant deviation between the two systems, the results are not comparable, and should be analyzed separately. In particular, the same system should be selected for monitoring the prognosis of MM.
Humans ; Immunoglobulin Light Chains ; Immunoglobulin kappa-Chains ; Immunoglobulin lambda-Chains ; Latex ; Multiple Myeloma/diagnosis*

Objective: To investigate the expression of histone methyltransferase G9a in gastric cancer tissues and its correlation to prognosis, and to observe the effect of G9a inhibitor on the proliferation and apoptosis of gastric cancer cells. Methods: The expression level of G9a in gastric cancer tissues and its correlation to prognosis were analyzed by using the Kaplan-Meier Plotter and Oncomine database. Human gastric cancer cell line SGC-7901 and MKN-45 were selected as study subject. The expression level of G9a protein was detected by Western blotting. The morphological change of gastric cancer cells after the treatment of G9a inhibitor BIX01294 was observed. CCK-8 proliferation experiment and plate colony formation assay were used to examine the proliferation ability and clone formation rate of gastric cancer cells. The changes of cell apoptosis were detected by Annexin-V staining. Results: G9a was highly expressed in gastric cancer tissues (P<0.01), and the high expression of G9a was positively correlated with poor prognosis of gastric cancer patients (P<0.01). After the treatment of BIX01294, the morphology of gastric cancer cells was changed, the volume of gastric cancer cells reduced, the intercellular connections disappeared, and even the apoptotic manifestations appeared, such as the shrinking,, becoming round and cast-off etc. BIX01294 could significantly inhibit the proliferation and colony formation but promote the apoptosis of gastric cancer cells (all P<0.05). Conclusion: Histone methyltransferase G9a was highly expressed in gastric cancer tissues, and its high expression level was positively correlated with poor prognosis. The proliferation of gastric cancer cells was obviously inhibited while the apoptosis was significantly promoted after inhibiting G9a expression.

Relative deficiency in production of glycoprotein hormone erythropoietin (Epo) is a major cause of renal anemia. This study planned to investigate whether the hypoxia-regulated system of Epo expression, constructed by fusing Epo gene to the chimeric phosphoglycerate kinase (PGK) hypoxia response elements (HRE) in combination with cytomegalovirus immediate-early (CMV IE) basal gene promoter and delivered by plasmid intramuscular injection, might provide a long-term physiologically regulated Epo secretion expression to correct the anemia in adenine-induced uremic rats. Plasmid vectors (pHRE-Epo) were synthesized by fusing human Epo cDNA to the HRE/CMV promoter. Hypoxia-inducible activity of this promoter was evaluated first in vitro and then in vivo in healthy and uremic rats (n = 30 per group). The vectors (pCMV-Epo) in which Epo expression was directed by a constitutive CMV gene promoter served as control. ANOVA and Student's t-test were used to analyze between-group differences. A high-level expression of Epo was induced by hypoxia in vitro and in vivo. Though both pHRE-Epo and pCMV-Epo corrected anemia, the hematocrit of the pCMV-Epo-treated rats exceeded the normal (P < 0.05), but that of the pHRE-Epo-treated rats didn't. Hypoxia-regulated system of Epo gene expression constructed by fusing Epo to the HRE/CMV promoter and delivered by plasmid intramuscular injection may provide a long-term and stable Epo expression and secretion in vivo to correct the anemia in adenine-induced uremic rats.
Anemia/blood/*therapy ; Animals ; Base Sequence ; Blood Urea Nitrogen ; Cell Hypoxia ; Creatinine/blood ; Erythropoietin/biosynthesis/*genetics/secretion ; Gene Expression Regulation ; Genes, Reporter ; Genetic Therapy ; HeLa Cells ; Humans ; Injections, Intramuscular ; Kidney/pathology ; Luciferases, Firefly/biosynthesis/genetics ; Molecular Sequence Data ; Plasmids/*genetics ; Promoter Regions, Genetic ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins/biosynthesis/genetics/secretion ; Response Elements ; Transcriptional Activation ; Uremia/blood/*therapy