To observe the effects of Niupo Zhibao Pellet, a compound traditional Chinese herbal medicine, on high-mobility group box-1 protein (HMGB1) expression in lung tissues of rats with endotoxin shock.

OBJECTIVE: To investigate the effect of Niupo Zhibao Pellet (NPZBP) on the expression of neuronal nitric oxide synthase (nNOS) in the brain of endotoxin-induced shock rats. METHODS: SD rats were randomly divided into normal control group, endotoxin-induced shock model group and NPZBP-treated group. Lipopolysaccharide (LPS) (1.5 mg/kg i.v.) and tsD-galactosamine (D-GalN) (100 mg/kg i.p.) were administered to the rats in endotoxin-induced shock model group, as well as to the rats in NPZBP-treated group after seven-day treatment, to induce the shock. The expression of nNOS in the brain of the rats in each of the 3 groups was measured by immunohistochemical methods. RESULTS: In the 3 groups, nNOS immuno-positive cells distributed widely in layer II, III, IV of the cerebral cortex, the molecular layer of hippocampus, the polymorphic layer of the dentate gyrus, the reticular formation of brain stem, and the molecular, granular and Purkinje cell layer of the cerebellar cortex. The number of immuno-positive cells in the NPZBP-treated group was slightly higher than that of the normal control group, and significantly lower than that of the model group (P<0.05) in many regions of the brain, including cerebral cortex, hippocampus, brain stem and cerebellar cortex. CONCLUSION: NPZBP can inhibit the over-expression of nNOS in wide area of the brain in endotoxin-induced shock rats.

Objective To investigate the effects of kidney-tonifying Plastrum Testudinis on o steogenesis of cultured rat mesenchymal stem cells(MSCs ).Methods MSCs were isolated from adult rats wi th density gradient separation meth od.Osteogenesis of MSCs in the culture a dded with different concentrations of serum containing Plastrum Testudinis was evalu-ated by detecting bone glaprotein(BGP)level and observing the morphologic al features of MSCs and by alkaline ph os-phatase(ALP)and Von Kossa immunohistochemical m ethods.Results Morphological examination showed t hat the MSCs attachment formed soon after seedin g ,and grew into colonies with the appearance of fibroblastic cells.Seru m containing Plastrum Testudinis promoted the expression of alkaline phosphatase(ALP)in MSCs ,and increased BGP level and the number of calcified deposition in dose -dependant manner,the difference being significant in comparison wi th the control groups(P

BACKGROUND: Stem cell differentiation potential is strongly correlated with culture condition. The alteration in scaffold material surface function, three dimensional (3D) structure, and addition of growth factors can control stem cell proliferation and differentiation.OBJECTIVE: To develop 3D macroporous scaffolds with optimal porosity and porous structure to provide a microenvironment that promotes the growth of multi-potent stem cells.DESIGN: Repetitive measurement.SETTING: Department of Anatomy, College of Basic Medicine, Guangzhou University of Chinese Medicine.MATERIALS: Healthy adult SD rats were provided by the Experimental Animal Center in Guangzhou University of Chinese Medicine. Chitosan and basic fibroblast growth factor (bFGF) were purchased from Sigma Corporation (St. Louis,MO).METHODS: The experiment was performed at the Department of Anatomy, College of Basic Medicine, Guangzhou University of Chinese Medicine from March 2003 to December 2006. Using a freeze-drying method, 3D macroporous scaffolds made of different ratios of chitosan-gelatin with bFGF were fabricated that could release bFGF with controlled porosity and porous structure. Bone marrow was obtained from the femur and tibia of SD rats, and mesenchymal stem cells (MSCs) were isolated, cultured and seeded on the scaffolds with bFGF. MSCs seeded on scaffolds with no bFGF served as control. The procedure during experiment was accorded with animal ethical requirements.MAIN OUTCOME MEASURES: 3D structure and release performance of the scaffolds were observed by ELISA and scanning electron microscope; the effect of 3D macroporous scaffolds that released bFGF on MSC growth and viability were observed by HE staining, MTT, cell counting and SEM.RESULTS: There was no significant difference in pore size between scaffolds with and without bFGF (P ＞ 0.05). Scaffolds with bFGF significantly improved MSC survival rate, promoted cell adhesion, proliferation, and viability compared with scaffolds without bFGF (P ＜ 0.05).CONCLUSION: The results suggest that 3D macroporous scaffolds with bFGF release improve MSC survival on scaffolds,and lay a foundation for its application in tissue engineering.

Objective To construct luciferase reporter vector containing full-length high-mobility group box 1 ( HMGB1, GenBank NM-010439) promoter for the screening of medicine. Methods The full-length HMGB1 promoter was amplified by polymerase chain reaction ( PCR) , and then was inserted into GV238 vector to construct plasmid GV238-HMGB1-P-Luc. GV238-HMGB1-P-Luc combined with internal reference plasmid pRL was co-transfected into Hela cells ( GV238-HMGB1-P-Luc group, which served as positive control group) . Plasmid pGL3-basic combined with pRL was co-transfected into Hela cells (pGL3-basic group, which served as negative control group) . Additionally, lipopolysaccharides ( LPS, 0.2 μg/mL) was used as the activator for the positive control group (LPS group), and then sodium butyrate (SB, 10 mmol/L) was used as the inhibitor for LPS group ( SB group) . At the end of experiment hour 24, luciferase activity was detected. Results The results of digestion, amplification, sequencing and identification showed that the full length of HMGB1 promoter was 2 140 bp, and the DNA sequence was correct, without mutation. Luciferase activity in GV238-HMGB1-P-Luc group was increased as compared with that of the pGL3-basic group ( P<0.05) . Luciferase activity in the LPS group was increased ( P<0.01, compared with that of GV238-HMGB1-P-Luc group) , and then was decreased after the administration of SB ( P<0.01, compared with that of the LPS group) . Conclusion A model of luciferase reporter vector containing HMGB1 promoter has been successfully constructed. Its activity can be increased by LPS, and then is in hibited by SB. The model can be used for further screening of medicine with the activities of regulating HMGB1 promoter.

The effects of RNAi-mediated gene silencing of LRIG3 expression on cell cycle and survival of human glioma cell line GLI 5 and the possible mechanisms were explored.The plasmids pGenesil2-LRIG3-shRNA l and pGenesil2-LRIG3-shRNA2 were transfected into GL 15 glioma cells respectively by using Metafectine,and the transfected cells that stably suppressed LRIG3 expression were selected by G418.The control cells were transfected with negative control shRNA.The changes in LRIG3 mRNA and protein levels were measured by RT-PCR and Western blot.The apoptosis rate and cell cycle were analyzed by flow cytometry.As compared with the negative shRNA-transfected GL15 cells,LRIG3 mRNA expression in GLI5 cells transfected with pGenesil2-LRIG3-shRNAl and pGenesil2-LRIG3-shRNA2 was silenced by 52.4%,63.8%,and LRIG3 protein expression was re-duced by 50.9% and 67.4% respectively.The LRIG3-specific siRNA transfected cells had higher proliferation rate than control cells.Cell cycle analysis showed that silencing LRIG3 increased the percentage of G2/M phase cells and the proliferation index significantly (P<0.01).Silencing LRIG3 could inhibit the apoptosis of GLl5 cells (P<0.05).These findings suggest that the siRNA targeting LRIG3 gene shows a dramatic inhibitory effect on RNA transcription and protein expression,then promoting the proliferation of GLI5 cells,arresting GLI5 cells in G2/M phase,and suppressing apoptosis ofGL15 cells.

Objective To summarize the clinical features of asymptomatic meningioma patients and evaluate the factors re-lated to surgical complications and prognosis,and to provide evidence for screening asymptomatic meningioma patients suitable for early surgical intervention.Methods The medical records of meningioma patients who underwent surgery from January 1st 2015 to December 31st 2020 at Neurosurgery Department of Tongji Hospital Affiliated to Tongji Medical College were retro-spectively reviewed.The clinical characteristics were compared between symptomatic and asymptomatic meningioma.Factors re-lated to the effects of surgery,postsurgical complications or patient prognosis were analyzed through Chi-squared test and multi-variate binary Logistic regression analysis.Results Elder age(HR:2.042;95％CI:1.002-4.098;P=0.021),smaller tumor size(HR:1.666;95％CI:1.009-3.857;P=0.014),intracranial superficial location(HR:2.221;95％CI:1.236-3.994;P=0.008)and no peritumoral brain edema(HR:8.917;95％CI:5.028-15.813;P＜0.01)were significant features of asymptomat-ic meningioma compared to those of symptomatic ones.The benefit of early surgery for asymptomatic meningioma was the a-chievement of higher total resection rate(88.6％)(P=0.035).Among the total resection cases,72％located in the intracranial superficial area,while merely 28％located in the intracranial deep area.Ninety-two percent of asymptomatic patients had re-turned to normal work and life at 6 months after operation.A parietal location was a significant factor indicating postsurgical complications(HR:3.351;95％CI:1.258-11.355;P=0.024),while elder age(≥60 years old)(HR:0.875;95％CI:0.825-0.999;P=0.041)was a significant factor indicating poor patient prognosis.Conclusion Asymptomatic meningioma is more common in elderly patients,usually located superficially in cranial cavity,with smaller size and without peritumoral edema.A more prominent total resection rate can be achieved in this type of meningioma.An early surgical resection would be recommen-ded as a proper treatment strategy.Tumor site and age are potential indicators for predicting postoperative complications and prognosis of patients.Comprehensive evaluation should be consider before surgery.

Objective To explore the correlation of molecular pathological grading with WHO grade 1 meningioma recurrence, malignant progression, and patients’ survival. Methods The medical records and paraffin-embedded tissues of patients with surgically resected WHO grade 1 meningioma were collected. The molecular pathological risk grading suggested by Maas et al. was adopted, and the patients were graded as low, intermediate, and high risk. Univariate log-rank test and multivariate Cox regression analyses were performed to determine the relationship between molecular risk grading and patient progression-free survival (PFS), malignant progression-free survival (MPFS), and overall survival (OS). Results Among 198 patients, 152 (76.8%) were graded as low risk, showing no 1p deletion; 42 (21.2%) patients were graded as intermediate risk, including 18 patients with 1p deletion, 10 patients with 1p combined with 6q deletion, and 14 patients with 1p combined with 14q deletion; and 4 (2%) patients were graded as high risk, including two patients with TERT promoter mutation, one patient with CDKN2A/B homozygous deletion, and one patient with 1p, 6p, and 14q combined deletion. Multivariate analysis showed that molecular risk grading was negatively associated with PFS (HR: 0.029, 95%CI: 0.011-0.080), MPFS (HR: 0.032, 95%CI: 0.004-0.274), and OS (HR: 0.074, 95%CI: 0.032-0.174; P<0.05). Conclusion The biological behavior of histological grade 1 meningiomas still exhibits heterogeneity, and further molecular pathological risk grading can more accurately reflect their biological behavior and evaluate patient prognosis.