Distinct Prognostic Factors of Ground Glass Opacity and Pure-Solid Lesion in Pathological Stage I Invasive Lung Adenocarcinoma.

Background: Ground glass opacity (GGO) is associated with favorable survival in lung cancer. However, the relevant evidence of the difference in prognostic factors between GGO and pure-solid nodules for pathological stage I invasive adenocarcinoma (IAC) is limited. We aimed to identify the impact of GGO on survival and find prognostic factor for part-GGO and pure-solid patients. Methods: Between December 2007 and August 2018, patients with pathological stage I IAC were retrospectively reviewed and categorized into the pure-GGO, part-GGO, and pure-solid groups. Survival curves were analyzed by the Kaplan-Meier method and compared by log-rank tests. Least absolute shrinkage and selection operator and Cox regression models were used to obtained prognostic factors for disease-free survival (DFS) and overall survival (OS). Results: The number of patients with pure-GGO, part-GGO, and pure-solid was 134, 540, and 396, respectively. Part-GGO patients with consolidation-tumor-ratio (CTR) > 0.75 had similar outcome to those with pure-solid nodules. In part-GGO patients, CTR was negatively associated with OS (P = 0.007) and solid tumor size (STS) was negatively associated with DFS (P < 0.001). Visceral pleural invasion (VPI) was negatively associated with OS (P = 0.040) and DFS (P = 0.002). Sublobectomy was negatively associated with OS (P = 0.008) and DFS (P = 0.005), while extended N1 stations examination was associated with improved DFS (P = 0.005) in pure-solid patients. Conclusions: Though GGO component is a positively prognostic factors of patients with pathological stage I IAC, a small proportion of GGO components is not associated with favorable survival. VPI, STS and CTR are the significant predictors for part-GGO patients. Sublobectomy, especially wedge resection should be used cautiously in pure-solid patients.

Analysis of risk factors of multiorgan failure after pericardiectomy for constrictive pericarditis.

BACKGROUND: We aimed to investigate risk factors of multiorgan failure following pericardiectomy. METHODS: This was a retrospective study of patients undergoing pericardiectomy between January 1994 and May 2021 at three hospitals. RESULTS: 826 patients were included in the study and divided into two groups: group with multiorgan failure (n = 86) and group without multiorgan failure (n = 740). There were 86 patients with multiorgan failure (86/826, 10.4%). There were 66 operative deaths (66/826, 8.0%). The causes of operative deaths were multiorgan failure, including cardiogenic shock + AKI + ventricular fibrillation (13/66), cardiogenic shock + AKI (35/66), cardiogenic shock + AKI + hepatic failure + septicemia (8/66), cardiogenic shock + AKI + respiratory failure (10/66). Univariate and multivariate analyses showed the factors associated with multiorgan failure, including male (P = 0.006), time between symptoms and surgery (P < 0.001), thickness of pericardium (P < 0.001), intubation time (P < 0.001), ICU retention time (P < 0.001), hospitalized time postoperative (P < 0.001), preoperative central venous pressure (P < 0.001), postoperative central venous pressure (P < 0.001), D0 fluid balance (P < 0.001), D2 fluid balance (P < 0.001), postoperative chest drainage (P < 0.001), preoperative LVEDD(P < 0.001), postoperative LVEDD (P < 0.001), surgical duration (P < 0.001), bleeding during operation (P < 0.001), serum creatinine 24 h after surgery (P = 0.042), serum creatinine 48 h after surgery (P < 0.001), fresh-frozen plasma (P < 0.001), packed red cells (P < 0.001), blood lactate (P < 0.001). CONCLUSION: In our study, incomplete pericardial dissection, fluid overload, delayed diagnosis and treatment are associated with multiorgan failure following pericardiectomy.

miR-219a-5p inhibits the pyroptosis in knee osteoarthritis by inactivating the NLRP3 signaling via targeting FBXO3.

PURPOSE: This work was to identify the function and mechanism of miR-219a-5p in regulating knee osteoarthritis (KOA). METHODS: Rat fibroblast-like synoviocytes (FLSs) were isolated to construct KOA cell model by lipopolysaccharide and adenosine triphosphate treatment. miR-219a-5p and FBXO3 expression in FLSs was modulated by transfection. Flow cytometry was executed to research FLSs apoptosis. Caspase-1 and IL-1ß expression in FLSs was researched by immunofluorescence. The binding between miR-219a-5p and FBXO3 was identified by dual luciferase reporter gene assay. KOA rat model and miR-219a-5p up-modulation KOA rat model were constructed. Step size of rats was analyzed. Knee joints of rats were experienced Safranin O-fast green staining to evaluate the knee joint injury. FBXO3, pyroptosis-associated proteins, and IL-1ß and IL-18 expression in FLSs and articular cartilage tissues of rats were assessed by Western blot, qRT-PCR and Enzyme-linked immunosorbent assay. RESULTS: KOA cell model had higher apoptosis percentage, expression of pyroptosis-associated proteins, and IL-1ß and IL-18 level. miR-219a-5p up-modulation decreased the above indicators, whereas miR-219a-5p down-modulation increased the above indicators. FBXO3 expression was directly repressed by miR-219a-5p. Loss of FBXO3 suppressed the above indicators. FBXO3 counteracted the suppression of miR-219a-5p on the above indicators. miR-219a-5p agomir attenuated knee joint injury, increased step size of KOA rats, and reduced FBXO3, pyroptosis-associated proteins and level of IL-1ß and IL-18 in the articular cartilage tissues of KOA rats. CONCLUSION: miR-219a-5p suppressed the pyroptosis in KOA by inactivating the NLRP3 signaling via targeting FBXO3, which might be a promising target for ameliorating KOA in the clinic.

Enhanced refractory organics removal by •OH and 1O2 generated in an electro-oxidation system with cathodic Fenton-like reaction.

Recently, carbon nanotubes coated carbon black and polytetrafluoroethylene (CNTs-C/PTFE) gas diffusion electrode was used as an air-cathode in an electro-oxidation (EO) system for effectively generating hydrogen peroxide (H2O2) through a 2-electron oxygen reduction reaction (ORR). This ORR-EO system not only lowered applied voltage and conserved energy, but the synergistic peroxone (O3/H2O2) reaction could increase hydroxyl radicals (•OH) generation for organics elimination. However, a significant proportion of H2O2 was left in the effluent of ORR-EO, which was a loss of resources and energy. In this study, a Fenton-like reaction for in-situ H2O2 decomposition to generate active oxidation species was inserted by introducing MnO2 into the cathodic catalyst layer, and the sole MnO2/CNTs-C/PTFE air-cathode could accomplish 90% of phenol degradation. When MnO2/CNTs-C/PTFE air-cathode combined with Ti/NATO anode in an ORR-EO system, all anodic oxidation, Fenton-like reaction, and peroxone took place to successfully generate •OH and singlet oxygen (1O2). Over 95% of TOC in phenol and landfill leachate bio-effluent was effectively eliminated, with 20% energy savings compared to the ORR-EO with CNTs-C/PTFE air cathode.

Analysis of Risk Factors of Early Mortality After Pericardiectomy For Constrictive Pericarditis.

BACKGROUND: We aimed to investigate risk factors of early mortality following pericardiectomy. METHODS: This was a retrospective study of patients undergoing pericardiectomy between January 1994 and May 2021 at The People's Hospital of Guangxi Zhuang Autonomous Region, Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine, and The People's Hospital of Liuzhou City. RESULTS: This study included 826 patients, who were divided into two groups: group with operative deaths (N = 66) and group without operative deaths (N = 760). There were 66 operative deaths (66/826, 8.0%). The causes of operative deaths were multiorgan failure (86/826, 10.4%). Preoperative CVP (P < 0.001), chest drainage (P < 0.001), surgical duration (P < 0.001), fluid balance postoperative day D2 (P < 0.001), and tuberculosis pericarditis (P = 0.001) in group with operative deaths were significantly higher than those in group without operative deaths. Univariate and multivariate analyses showed that factors associated with operative deaths include male (P < 0.001), age (P < 0.001), ICU retention time (P < 0.001), postoperative hospitalization time (P < 0.001), preoperative central venous pressure (P = 0.018), postoperative central venous pressure (P < 0.001), D0 fluid balance (P < 0.001), D2 fluid balance (P < 0.001), postoperative chest drainage (P = 0.029), surgical duration (P = 0.003), serum creatinine baseline (P = 0.002), serum creatinine 24h after surgery (P < 0.001), serum creatinine 48h after surgery (P < 0.001), blood lactate (P < 0.001), and tuberculosis pericarditis (P = 0.033). CONCLUSION: In our study, incomplete pericardial dissection, fluid overload, and tuberculosis pericarditis are associated with operative deaths following pericardiectomy.

Incomplete Pericardial Dissection, Fluid Overload, Delayed Diagnosis And Treatment, And Tuberculosis Pericarditis Are Associated With Low Cardiac Output Syndrome Following Pericardiectomy.

BACKGROUND: We aimed to investigate risk factors of LCOS following pericardiectomy. METHODS: This was a retrospective study of patients undergoing pericardiectomy at three hospitals between January 1994 and May 2021. RESULTS: A total of 826 patients were divided into two groups: group with LCOS (N = 126) and group without LCOS (N = 700). The incidence of postoperative LCOS was 15.3%. There were 66 operative deaths (8.0%). Univariable and multivariable analyses showed that factors are associated with LCOS, including postoperative LVEDD (P < 0.001), preoperative LVEDD (P < 0.001), time between symptoms and surgery (P < 0.001), thickness of pericardium (P < 0.001), intubation time (P = 0.002), hospitalized time postoperative (P < 0.001), preoperative central venous pressure (P = 0.016), postoperative central venous pressure (P = 0.034), D0 fluid balance (P = 0.019), D2 fluid balance (P = 0.017), postoperative chest drainage (P < 0.001), surgical duration (P < 0.001), bleeding during operation (P = 0.001), serum creatinine 24h after surgery (P < 0.001), serum creatinine 48h after surgery (P = 0.017), fresh-frozen plasma (P = 0.005), packed red cells (P = 0.006), and tuberculosis pericarditis (P = 0.026). CONCLUSION: In our study, incomplete pericardial dissection, fluid overload, delayed diagnosis and treatment, and tuberculosis pericarditis are associated with LCOS following pericardiectomy.

[Generation and phenotypic characterization of S100A9 gene knockout mice by CRISPR/Cas9-mediated gene targeting].

S100 calcium binding protein A9 (S100A9) is involved in a variety of biological processes such as inflammation and tumor cell migration and invasion regulation. The purpose of this study was to construct S100A9 gene-edited mice by using CRISPR/Cas9 technology, thereby providing an animal model for exploring the biological functions of this gene. According to the S100A9 gene sequence, the single-stranded small guide RNA (sgRNA) targeting exons 2 and 3 was transcribed in vitro, and a mixture of Cas9 mRNA and candidate sgRNA was injected into mouse fertilized eggs by microinjection. Early embryos were obtained and transferred to surrogate mice, and F0 mice were obtained and identified by PCR identification and gene sequencing. F0 mice were further mated with wild-type C57BL/6 mice to obtain F1 heterozygous mice, and then homozygous offspring were obtained through F1 mice self-crossing. Real-time PCR, Western blot and immunohistochemistry (IHC) were used to verify the expression and distribution of S100A9. In order to observe the pathological changes of mouse lung tissue using HE staining, an allergic asthma model was induced by ovalbumin from chicken egg white (OVA). The results showed that the 2 492 bp of exons 2, 3 of the S100A9 gene was successfully knocked out, and S100A9-/- mice with stable inheritance were obtained. Furthermore, it was found that S100A9 gene was highly expressed in the lung and spleen of wild-type mice. The expression of S100A9 mRNA and protein was not detected in the lung and spleen of S100A9-/- mice. However, compared with wild-type mice, the lungs of S100A9-/- mice showed a significantly worse inflammatory phenotype, and the proportion of eosinophils in bronchoalveolar lavage fluid (BALF) was significantly increased in response to the treatment of OVA. These results suggest we have successfully constructed a new strain of S100A9-/- mice, and preliminarily confirmed that the lack of S100A9 function can aggravate airway inflammation in asthmatic mice, providing a new mouse model for further study of S100A9 gene function.

Structural determination and in vitro tumor cytotoxicity evaluation of five new cycloartane glycosides from Asplenium ruprechtii Sa. Kurata.

Five new cycloartane glycosides, named aspleniumside A - E, were discovered and characterized by re-investigated the remaining extracts of the whole plant of Asplenium ruprechtii Sa. Kurata, a famous folk medicine for treating thromboangitis obliterans in China, Japan, and Korea. Compounds 3-5 possessed the 9,19-seco-cycloartane-9,11-en triterpene aglycone with 3,7(or 23),24,25,30-highly oxidized methylene, methylene or quaternary carbons, that was found in this species for the first time. The stereo-chemistry of all new compounds were fully discussed by extensive analysis of the 1D and 2D NMR data, and comparisons with those data of known compounds. 24R configuration was determined here which indicated the different growing areas of the same species could influence the secondary metabolic behavior, leading to the differences in chemical composition. All glycoside groups were determined as ß-d-glucopyranosyl by 1H coupling constant of anomeric protons and co-TLC of the acid hydrolysate with d-glucose. All the cycloartane glycosides were evaluated against HL-60 and HepG2 cells for cytotoxicity, compounds 1-3, showed potential cytotoxicity with the IC50 in range of 18-60 µM, while the standard sorafenib showed IC50 value of 10.61 ± 0.43 and 13.43 ± 1.12 µM against HL-60 and HepG2, respectively. The results attained in this study indicated that cycloartane glycosides should be the cytotoxicity substance in A. ruprechtii Sa. Kurata, and had the potential to be developed as tumor cytotoxicity agent applied in clinic.

Enhanced electro-oxidation/peroxone (in situ) process with a Ti-based nickel-antimony doped tin oxide anode for phenol degradation.

Recently, a novel proof-of-concept oxygen reduction reaction (ORR) based electro-oxidation (EO) process has been developed, which was accomplished by integrating anodic electrochemical oxidation coupled with an in situ electro-peroxone process, by harnessing the anodic by-product O3 reacted with ORR cathode generated H2O2. To further enhance EO coupled in situ electro-peroxone, a nickel and antimony doped tin oxide anodic catalyst layer, namely NATO, was fabricated on Ti mesh to improve anodic oxidation and reinforce the generation of O3, thus promoting in situ Electro-peroxone. As a result, O3 generation rate was enhanced by 12.6%. Complete phenol, as a model organic compound, and 95% of TOC removal were achieved, respectively, during ORR-EO. Through kinetics and instrument analysis, results show that the amount of intermediates accumulated during phenol degradation was much less in this Ti/NATO based ORR-EO system than in a traditional EO system. Moreover, 35.7% of the energy consumption was saved for ORR-EO, owing to its reduced applied voltage and the enhanced in situ electro-peroxone process.

Metal trabecular bone reconstruction system better improves clinical efficacy and biomechanical repair of osteonecrosis of the femoral head than free vascularized fibular graft: A case-control study.

In this study, we aim to compare and analyze the biomechanical repair and clinical efficacy of osteonecrosis of the femoral head (ONFH) with the use of metal trabecular bone reconstruction system and free vascularized fibular graft. The study enrolled 66 adult patients from medical records of nontraumatic ARCO 2A-3B stage ONFH. A simple ONFH model without surgical treatment was established in 13 cases, 29 cases were treated with metal trabecular bone reconstruction system, and 24 cases were treated with free vascularized fibular graft. Computer-recognized and extracted femur outlines were imported, and three-dimensional reconstructions were performed. The stress concentration and stress peak value were analyzed, and the Harris score, visual analog scale pain score, and operation status of the above patients were compared. Finally, quality of life assessment was performed using SF-36 scale. Metal trabecular bone reconstruction system provided less operation time, blood loss, and the total length of postoperative hospital stay than free vascularized fibular graft. Metal trabecular bone reconstruction system promoted bone reconstruction, increased bone mineral density and Harris score. The total clinical effective rate of young patients (20-40 years) was higher than that of older patients (41-60 years). Metal trabecular bone reconstruction system provided higher physical component summary, mental component summary, and role/social component summary than free vascularized fibular graft. This study demonstrates that both metal trabecular bone reconstruction system and free vascularized fibular graft can prevent or delay the progression of ONFH, while metal trabecular bone reconstruction system is a better choice because of better short-term clinical efficacy.

An anti-inflammatory C-stiryl iridoid from Camptosorus sibiricus Rupr.

A new iridoid glycoside, named camptoside (1), together with three known compounds as dehydrodiconiferyl alcohol-9'-O-ß-d-glucopyranoside (2), aesculetin (3) and vajicoside (4), have been isolated from Camptosorus sibiricus Rupr. (Aspleniaceae). Their structures were established on the basis of spectroscopic analysis, especially 1D- and 2D-NMR data, and by comparison of their spectroscopic and physical data with those reported in the literature. Compounds 1-3 exhibited inhibitions of nitric oxide production in lipopolysaccharide-induced RAW 264.7 macrophages with IC50 values of 11.2, 8.3 and 9.4 µM, respectively.

Cuff-leak test combined with interventional bronchoscopy benefits early extubation for patients who received tarp surgery.

PURPOSE: This study explored the performance characteristics of a cuff-leak test (CLT) combined with interventional fiberoptic bronchoscopy (FBS) for evaluating whether early nasoendotracheal extubation was possible for patients who had received transoral atlantoaxial reduction plate (TARP) internal fixation surgery. METHODS: 318 patients who underwent surgery were retrospectively analyzed (between January 2006 and December 2012). Extubation was performed by conventional approach (CA group, until December 2008) and improved approach (IA group, from January 2009) including CLT and an interventional FBS procedure. The extubation success within 1-3 days after surgery, incidence of postextubation stridor and tracheal reintubation were examined. RESULTS: More IA-treated patients experienced extubation during the first 2 days than those CA-treated, median extubation time was 3 (2, 3) days in the CA group and 2 (1, 2) days in the IA group (all P < 0.01). The incidence of stridor and reintubation was 5.69 and 0.57 % in IA and 11.98 and 4.93 % in CA, respectively (both P < 0.05). For the CLT-positive patients in the IA group that remained intubated until day 3-4, interventional FBS was applied for safe extubation and achieved 100 % success. CONCLUSION: Early extubation through IA is safe and interventional FBS assists successful extubation for CLT-positive patients who underwent TARP surgery.

The decreased N6-methyladenine DNA modification in cancer cells.

N6-methyladenine (6 mA) is a recently characterized DNA modification in mammalian genomes, although its biological importance remains to be resolved. Using a highly sensitive HPLC/MS/MS approach, here we report regulation of 6 mA modification in mammalian cells. To these aspects, down-regulation of 6 mA modification was first characterized in human cancer cells and tissues, relative to their normal controls. In contrast to the relative stable 5 mC modification, a dramatic decrease of 6 mA modification was found, showing that 6 mA is the most regulated DNA modification in cancers. In addition to the regulation in cancer cells, a hundreds-fold increase of 6 mA modification was found for in vitro cultured human cells, relative to the in vivo cells. This up-regulation was also confirmed with in vitro cultured mouse cells. Taken together, our study revealed distinct 6 mA modification profiles in the cancer and cultured cells. Considering its distinct regulation from that of 5 mC, our study suggests that 6 mA DNA modification may play a crucial role in cell fate transition of mammalian cells.

[Overexpression of Sox9 gene by the lentiviral vector in rabbit bone marrow mesenchymal stem cells for promoting the repair of cartilage defect].

OBJECTIVE: To study the overexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells for repairing articular cartilage injury in vivo. METHODS: Rabbit bone marrow mesenchymal stem cells (BMSCs) were transduced with lentivirus vector containing Sox9 gene and then cartilage specific molecule was detected by RT-PCR in vitro. Total 48 knee joints of 24 mature New Zealand white rabbits were randomly divided into 3 groups according to different defect treatment. After animals anesthesia,a full-thickness cylindrical cartilage defect of 4 mm diameter and 3 mm deep was created in the patellar groove using a stainlesssteel punch. Meanwhile, the transfected cells were implanted to repair the rabbit model with full-thickness cartilage defects. Cartilage defects tissue was observed with light microscope, electron microscope, HE and immunohistochemistry staining to assess the repair of defects by the complex at 6 weeks or 12 weeks after the implantation. RESULTS: At 3 days after the transfection, Sox9 gene expression was highest and Sox9 gene expression decreased with the increase of time. At 3 days after the transfection, the expression of collagen type II began and reached the peak at 14 days. It showed that the bone marrow mesenchymal stem cells went into chondrogenic differentiation after transfected by Sox9 gene. Histological observation showed that at 6 weeks after the operation, the defects in the experimental group was filled with hyaline like cartilage tissue, 12 weeks after operation,the defects of cartilage and subchondral bone had satisfactory healing. Both at 6 and 12 weeks postoperatively, the defects were filled with fibrous tissues in control groups. Meanwhile, immunohistochemical staining of sections with type II collagen antibodies showed the proteins in the regenerated tissue stained positive for type II collagen and stronger than the control groups. The histological scoring system indicated that the cartilage repair of experiment groups were better than the two control groups with statistical significances. CONCLUSION: Overexpression of Sox9 gene on rabbit bone marrow mesenchymal stem cells (BMSCs) promote the repair of cartilage defect.

Nonionic surfactant greatly enhances the reductive debromination of polybrominated diphenyl ethers by nanoscale zero-valent iron: mechanism and kinetics.

Nanoscale zero-valent iron (nZVI) has been considered as an effective agent for reductive debromination of polybrominated diphenyl ethers (PBDEs). But the high lipophilicity of PBDEs will hinder their debromination owing to the inefficient contact of PBDEs with nZVI. In this study, different ionic forms of surfactants were investigated aiming to promote PBDE debromination, and the beneficial effects of surfactant were found to be: nonionic polyethylene glycol octylphenol ether (Triton X-100, TX)>cationic cetylpyridinium chloride (CPC)>anionic sodium dodecyl benzenesulfonate (SDDBS). Except for with SDDBS, the promotion effect for PBDE debromination was positively related to the surfactant concentrations until a critical micelle concentration (CMC). The debromination process of octa-BDE and its intermediates could be described as a consecutive reaction. The corresponding rate constants (k) for the debromination of parent octa-BDE (including nona- to hepta-BDEs), the intermediates hexa-, penta-, and tetra-BDEs are 1.24 × 10(-1) h(-1), 8.97 × 10(-2) h(-1), 6.50 × 10(-2) h(-1) and 2.37 × 10(-3) h(-1), respectively.

[Reductive debromination of polybrominated diphenyl ethers in aquifier by nano zero-valent iron: debromination kinetics and pathway].

Nano-zerovalent iron (nZVI) approach is effective in the debromination of polybrominated biphenyl ethers (PBDEs). The kinetics and degradation pathway are the key issues to understand the PBDEs degradation mechanisms. In this study, nZVI, synthesized through liquid phase reduction method, coupled with Triton X-100, could completely debrominate the highly brominated congeners of a commercial octa-BDEs mixture within 46 h. The debromination of octa-BDEs could be described by means of pseudo-first-order kinetics with the reaction constant (k) of 0.106 h(-1). In case of lacking the PBDE standards, an effective approach has been developed to determine the unknown PBDE congeners using the quantitative-structure retention relationship (QSRR) model. The retention time of all 39 PBDE congeners in a standard mixture was firstly analyzed with gas chromatography coupled with an electron capture detector (GC-ECD), and the relative retention time (RRT) for each standard was obtained after normalizing the RT by the average RT of BDE47 and BDE183. Then a QSRR model was developed by fitting the RRT of each PBDE congener and its specific RRT index. The debromination products of octa-BDEs were identified using this QSRR model and the degradation pathway of octa-BDEs was elucidated. The results showed that in the stepwise reductive debromination process of PBDEs by nZVI, meta-debromin was facile to be degraded.

Visualizing cell state transition using Raman spectroscopy.

System level understanding of the cell requires detailed description of the cell state, which is often characterized by the expression levels of proteins. However, understanding the cell state requires comprehensive information of the cell, which is usually obtained from a large number of cells and their disruption. In this study, we used Raman spectroscopy, which can report changes in the cell state without introducing any label, as a non-invasive method with single cell capability. Significant differences in Raman spectra were observed at the levels of both the cytosol and nucleus in different cell-lines from mouse, indicating that Raman spectra reflect differences in the cell state. Difference in cell state was observed before and after the induction of differentiation in neuroblastoma and adipocytes, showing that Raman spectra can detect subtle changes in the cell state. Cell state transitions during embryonic stem cell (ESC) differentiation were visualized when Raman spectroscopy was coupled with principal component analysis (PCA), which showed gradual transition in the cell states during differentiation. Detailed analysis showed that the diversity between cells are large in undifferentiated ESC and in mesenchymal stem cells compared with terminally differentiated cells, implying that the cell state in stem cells stochastically fluctuates during the self-renewal process. The present study strongly indicates that Raman spectral morphology, in combination with PCA, can be used to establish cells' fingerprints, which can be useful for distinguishing and identifying different cellular states.

[Gene expression of bone mesenchymal stem cells transduced by the lentiviral vector of SOX9 gene].

OBJECTIVE: To construct one lentiviral vector containing mouse SRY-related high mobility group-box gene 9 (SOX9) and transfect the murine bone mesenchymal stem cells (mBMSCs) in vitro and observe the expression of target gene. METHODS: RNA from the vectors containing mouse SOX9 gene were extracted and SOX9 genes were amplified by reverse transcription-Polymerase Chain Reaction (RT-PCR). The SOX9 genes were connected into lentiviral vectors pGC-FU. Then pGC-FU-SOX9 transduced into 293T cells to produce recombinant lentivirus called as Lenti-SOX9-EGFP. mBMSCs were transfected. The expression of target gene was detected by immunofluorescence, RT-PCR and Western Blot. RESULTS: Lenti-SOX9-EGFP was recombined successfully and transduced efficiently into mBMSCs. The expression of SOX9 gene was confirmed by RT-PCR and Western Blot. CONCLUSION: Lentiviral vector of mouse SOX9 gene can transfect successfully into mBMSCs. Meanwhile, SOX9 gene may be expressed in mBMSCs. This will provide the target cells for the following study about SOX9 gene repairing cartilage injury.

[Gene expression of bone mesenehymal stem cells transduced by the lentiviral vector of SOX9 gene knockdown].

OBJECTIVE: To construct one lentiviral vector containing mouse SRY-related silencing group--box gene 9 (SOX9) and to transfect murine bone mesenehymal stem cells (mBMSCs) in vitro and observe the expression of target gene. METHODS: RNA inteference target sequence was designed in connectin with mice SOX9 gene sequence. The double strands DNAoligo containing interference sequence were synthesized and cloned into lentivirus vector. The siRNA lentiviral vector with SOX9 gene silencing was constructed and identified, which was transfected into rat bone mesenehymal stem cells. The expression of target gene was detected by immunofluorescence, RT-PCR and Western blot. RESULTS: Lenti-SOX9-siRNA-EGFP was recombined successfully and transduced efficiently into mBMSCs. The expression of SOX9 gene silencing was confirmed by RT-PCR and Western blot. CONCLUSION: Mouse SOX9 gene silencing by RNA interference and Lentiviral vector can transfected successfully into mBMSCs. Meanwhile,SOX9 gene may be silenced in SOX9 transduced mBMSCs. This will provide target cells for the following study about SOX9 gene respairing cartilage injury.