OBJECTIVETo establish the quantitative method of notoginsenoside R1, ginsenoside Rg1 and Rb1 in Radix Notoginseng and its preparation Xuesaitong injection by HPLC-ELSD.

METHODThe column was packed with 5 microm Diamonsil C18 stationary phase. The mobile phase consisted of acetonitrile-water, eluted in gradient mode. The temperature of drift tube was 105 degrees C and the nebulizer nitrogen flow rate was 2.9 L x min(-1).

RESULTThe linear ranges of the three components were 0.456-2.25 microg, 1.47-7.38 microg and 1.20-6.03 microg respectively. The average recoveries of the three components in Radix Notoginseng were 97.1% (RSD 1.9%), 96.8% (RSD 2.0%), 97.0% (RSD 2.2%) respectively; in Xuesaitong Injection were 98.7% (RSD 1.9%), 98.5% (RSD 1.8%), 98.1% (RSD 1.4%) respectively.

CONCLUSIONIt was proved that the method was reliable, simple, and precise, that could be used for quality control.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Ginsenosides ; analysis ; Injections ; Panax ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control

bThis study explored whether the transplantation of modified marrow stromal cells (MSCs) has angiogenic effects in a left middle cerebral artery occlusion infarction/reperfusion (MCAO I/R) rat model and preliminarily examined the mechanism of angiogenesis following cerebral infarction. MSCs were isolated by using a direct adherent method and cultured. Vascular endothelial growth factor (VEGF) was transfected into MSCs by employing the liposome transfection. The transfection efficiency was measured by the optical density method. The protein expression of VEGF gene before and after transfection was measured by Western blotting. SD rat model of transient occlusion of the left middle cerebral artery was established by using an approach of intra-luminal occlusion. Tetrazolium (TTC) and HE staining were performed to observe the cerebral infarction. ELISAs were used to measure the levels of VEGF in the rat cerebral tissues. The expression patterns of angiopoietin-2 (Ang-2) and CD34 in cells surrounding the area of infarction were immunohistochemistrically observed. Ang-2 protein expression in the tissue surrounding the area of infarction was measured by Western blotting. VEGF expression in the MSCs increased after transfection at a rate of approximately 28%±3.4%. ELISA showed that the expression of VEGF in the cerebral tissue was significantly increased after induction of infarction, peaking on the 4th day and decreasing to the levels of the sham surgery group (normal) within 7 to 10 days. The VEGF level was significantly higher at each time point in the VEGF-MSC and MSC groups compared to the model group. Moreover, the VEGF level was higher in the VEGF-MSC group than in the MSC group and stayed relatively high until the 10th day. The immunohistochemical results showed that 10 days after the infarction, the number of Ang-2 and CD34-expressing cells in the area surrounding the infarction was significantly higher in the VEGF-MSC group and the MSC group compared to the model group. Moreover, the VEGF level was higher in the VEGF-MSC group than the MSC group. A similar trend in Ang-2 protein expression was revealed by Western blotting. In the MCAO rat model transfected with modified MSCs over-expressing VEGF, compared to the MSC transplantation group, the concentration of VEGF was significantly increased in the brain tissue after cerebral infarction. In addition, the level of Ang-2 was up-regulated, with angiogenesis promoted, the blood supply to the areas surrounding the cerebral infarction increased, and neurological function improved. We are led to speculate that the synergistic effects of VEGF and Ang-2 may be responsible for the angiogenesis following cerebral infarction.

bThis study explored whether the transplantation of modified marrow stromal cells (MSCs) has angiogenic effects in a left middle cerebral artery occlusion infarction/reperfusion (MCAO I/R) rat model and preliminarily examined the mechanism of angiogenesis following cerebral infarction. MSCs were isolated by using a direct adherent method and cultured. Vascular endothelial growth factor (VEGF) was transfected into MSCs by employing the liposome transfection. The transfection efficiency was measured by the optical density method. The protein expression of VEGF gene before and after transfection was measured by Western blotting. SD rat model of transient occlusion of the left middle cerebral artery was established by using an approach of intra-luminal occlusion. Tetrazolium (TTC) and HE staining were performed to observe the cerebral infarction. ELISAs were used to measure the levels of VEGF in the rat cerebral tissues. The expression patterns of angiopoietin-2 (Ang-2) and CD34 in cells surrounding the area of infarction were immunohistochemistrically observed. Ang-2 protein expression in the tissue surrounding the area of infarction was measured by Western blotting. VEGF expression in the MSCs increased after transfection at a rate of approximately 28%±3.4%. ELISA showed that the expression of VEGF in the cerebral tissue was significantly increased after induction of infarction, peaking on the 4th day and decreasing to the levels of the sham surgery group (normal) within 7 to 10 days. The VEGF level was significantly higher at each time point in the VEGF-MSC and MSC groups compared to the model group. Moreover, the VEGF level was higher in the VEGF-MSC group than in the MSC group and stayed relatively high until the 10th day. The immunohistochemical results showed that 10 days after the infarction, the number of Ang-2 and CD34-expressing cells in the area surrounding the infarction was significantly higher in the VEGF-MSC group and the MSC group compared to the model group. Moreover, the VEGF level was higher in the VEGF-MSC group than the MSC group. A similar trend in Ang-2 protein expression was revealed by Western blotting. In the MCAO rat model transfected with modified MSCs over-expressing VEGF, compared to the MSC transplantation group, the concentration of VEGF was significantly increased in the brain tissue after cerebral infarction. In addition, the level of Ang-2 was up-regulated, with angiogenesis promoted, the blood supply to the areas surrounding the cerebral infarction increased, and neurological function improved. We are led to speculate that the synergistic effects of VEGF and Ang-2 may be responsible for the angiogenesis following cerebral infarction.
Angiopoietin-2 ; genetics ; metabolism ; Animals ; Bone Marrow ; metabolism ; pathology ; Cerebral Infarction ; genetics ; metabolism ; pathology ; Male ; Neovascularization, Pathologic ; genetics ; pathology ; Rats ; Rats, Sprague-Dawley ; Stromal Cells ; metabolism ; pathology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

INTRODUCTIONBased on the estimate results of the capacity and preparedness of Beijing hospitals to respond to pandemic influenza, using flu surge model to evaluate its applicable hypothesis and to provide government with sentient strategy in planning pandemic influenza. Through collection of medical resources information, we calculated the possible impaction on hospitals by Flu Surge model and explored the applicable hypothesis in model operation through a questionnaire, direct observation and group discussion in 3 hospitals in Beijing. Based on flu surge model estimation during a 6-week epidemic from a pandemic virus with 35% attack rate, Beijing would have had an estimation of 5 383 000 influenza illnesses, 2 691 500 influenza outpatients, 76 450 influenza hospitalizations and 14 508 excess deaths. For a 6-week period with 35% attack rate, there would be a peak demand for 8% of beds, 210% of ICU beds, and 128% of ventilators estimated. Outpatients in different level hospital were quite disproportionated with 1742/ hospital/day, 650/hospital/day, and 139/hospital/day respectively. The sampled health workers had a mastery of 63.4% of the total knowledge and skills of diagnosing and treating of influenza, 73.5% of them washed their hands and 63.5% used PPE correctly. The total beds capacity, medical beds capacity and respiratory medical beds capacity would increase 8%, 35% and 128% respectively.

CONCLUSIONThe estimation results could be referenced when planning the pandemic strategy, but the results should be treated objectively when considering the hypothesis and practical situation in this model being used.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Disease Outbreaks ; statistics & numerical data ; Female ; Hospital Bed Capacity ; Hospital Planning ; Hospitalization ; statistics & numerical data ; Humans ; Infant ; Infant, Newborn ; Influenza, Human ; epidemiology ; Male ; Middle Aged ; Models, Statistical ; Surge Capacity ; Young Adult

OBJECTIVESTo construct a recombinant adenoviral vector carrying antisense matrix metalloproteinase-2 (MMP2) and to study its inhibitory effects on the invasiveness and migratory capacity of hepatocellular carcinoma (HCC) cell line HepG2 in vitro.

METHODSTotal RNA was extracted from HCC. Then a 500 bp fragment at the 5' end of the human MMP2 cDNA sequence was synthesized by polymerase chain reaction (PCR) and was reversely inserted into the multiclone site (MCS) of the shuttle plasmid pAdTrack-CMV. With the resultant plasmid and the backbone plasmid pAdEasy-1, the homologous recombination took place in the E.coli BJ5183 and the recombinant adenoviral plasmid carrying the antisense MMP2 gene was constructed. The adenovirus (Ad-MMP2AS) was packaged and amplified in the HEK 293 cells and the viral titer was checked by GFP. Using the Boyden chamber model, the influence of Ad-MMP2AS on the invasion ability of HepG2 cells was determined in vitro.

RESULTSThe recombinant adenovirus vector carrying antisense MMP2 was constructed successfully and a strong green fluorescence was observed in HepG2 cells under a fluorescence microscope. The viral titer was 1 x 10(8); Ad-MMP2AS can effectively inhibit the penetrating capacity of HepG2 cells through Matrigel in vitro.

CONCLUSIONThe recombinant adenovirus with antisense MMP2 can effectively inhibit the invasiveness and migratory capacity of HepG2 in vitro and may have potential in treating HCC.

Adenoviridae ; genetics ; Carcinoma, Hepatocellular ; pathology ; Genetic Vectors ; Humans ; Liver Neoplasms ; pathology ; Matrix Metalloproteinase 2 ; biosynthesis ; genetics ; pharmacology ; Neoplasm Invasiveness ; Oligonucleotides, Antisense ; biosynthesis ; genetics ; pharmacology ; RNA, Antisense ; genetics ; pharmacology ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology ; Tumor Cells, Cultured

Objective:To explore the clinical features, follow-up characteristics and prognosis of rheumatic disease complicated with pulmonary arterial hypertension (PAH) in children, and to provide support for its clinical diagnosis and treatment.Methods:A retrospective analysis was conducted on the data of rheumatic 24 patients complicated with PAH hospitalized in the Department of Rheumatology and Immunology, Children′s Hospital Affiliated to the Capital Institute of Pediatrics, Department of Rheumatology and Immunology, Jiangxi Children′s Hospital, Department of Pediatrics Ⅰ, the First Affiliated Hospital of Zhengzhou University and Department of Pediatrics, the Affiliated Hospital of Inner Mongolia Medical University from January 2013 to June 2022.The rheumatic patients complicated with PAH were followed up by telephone on June 30, 2022, and their clinical symptoms, treatment, follow-up, and prognosis data were collected.According to different treatment methods, the patients were divided into different clinical subgroups. The change of PAH was analyzed. The t-test was used for comparison between groups. P<0.05 was statistically significant. Results:A total of 24 cases were enrolled, with 7 males and 17 females.The average onset age of PAH was (10.97±3.79) years old.The median duration of PAH was 6.00 (32.20) months.The average pulmonary artery pressure was (51.71±17.66) mmHg(1 mmHg=0.133 kPa). There were 9 cases of systemic lupus erythematosus, 5 cases of Takayasu′s arteritis, 3 cases of juvenile dermatomyositis, 3 cases of undifferentiated connective tissue disease, 2 cases of systemic juvenile idiopathic arthritis, 1 case of Behcet′s disease, and 1 case of Kawasaki disease.Among 24 cases, the common symptoms were fever (14 cases), fatigue (10 cases) and dyspnea (7 cases). Of the 24 cases, 10 cases were complicated with hydropericardium, 9 cases with valve regurgitation, and 5 cases with decreased systolic and/or diastolic function.Lung changes were observed in 17 cases.Eleven cases were tested for B-type natriuretic peptide (BNP), and the BNP levels were all elevated in them (11 cases), with a median BNP of 3 073 (10 645) ng/L.After the first occurrence of PAH, 12 cases were treated with Methylprednisolone therapy, 10 cases received Cyclophosphamide therapy, and 2 cases who were both systemic lupus erythematosus, underwent blood purification.In the treatment of PAH, 11 cases were treated with pulmonary artery pressure reduction, and 7 of the 11 cases took PAH-targeted drugs.The mean decrease of the average pulmonary artery pressure in children receiving the targeted therapy[(44.80±24.08) mmHg] was significant higher than that in children not receiving the targeted therapy [(16.15±17.25) mmHg] ( t=2.661, P=0.016). Twenty children were reexamined and/or followed up, and the average course of PAH at the telephone follow-up was (36.29±26.67) months.The pulmonary arterial hypertension in 6 cases completely recovered, with median recovery time of 8.00 (13.47) months, but 2 of them died after the complete recovery.The pulmonary arterial hypertension improved in 11 children, 1 of whom died and the remaining children were in stable condition.The pulmonary arterial hypertension worsened in 2 children, 1 of them improved previously but aggravated recently, and the other child did not monitor pulmonary artery pressure and died during telephone follow-up. Conclusions:Rheumatic diseases complicated with PAH are rare and most often diagnosed in severe rheumatic children.It can lead to death, and is commonly accompanied by notably elevated BNP levels.The patients who have early PAH detection, intensive treatment of the primary disease, symptomatic and targeted pulmonary artery pressure reduction show a better prognosis.

To identify human monoclonal HIV-l-neutralizing antibodies from an HIV-1 CRF07BC specific phage display antibody library by cell-based screening. 293T cells were transfected by pCH064. 2-Env plas mid and then used to biopan the phage antibody library. The positive phage clones were screened by cell based ELISA and sequenced for the variable region of heavy (VH) and light (VL) chains. The expressed Fabs were purified by Ni(+2) -NTA column and analyzed by SDS-PAGE. The cell- and gp120 protein-based ELISA as well as flow cytometry were used to measure Fab's binding activity. The neutralizing activity of Fabs was assessed by HIV-1 pseudoviruses. After 4-round biopanning, the binding phages to transfected cells were enriched about 650-folds. A total of 28 positive clones were screened out by cell ELISA and sequence analysis identified 5 different Fabs possessing unique VH and VL (2801, 2837, 2863, 2870 and 2920). Interestingly, these Fabs reacted with the Env-transfected 293T cells but not soluble gp120 proteins, suggesting that they might target conformation-dependent epitopes presenting on viral Env complex. We found that three Fabs (2801, 2863, 2870) exhibited potent neutralizing activity against CRF07_BC isolate CH120. 6 with IC50 of 2.24, 0.89 and 3.09 microg/mL respectively, and that 2801 and 2863 cross-neutral ized the subtype B isolate SF162 at IC50 of 0.69 and 3.52 microg/mL respectively. In conclusion, the HIV-1 Env-transfected 293T cells can be used to efficiently enrich and screen the phage antibody library and isolate human monoclonal HIV-1-neutralizing Fabs that target the Env complex-dependent conformational epitopes. Therefore, our studies provide a powerful platform for exploring the mechanism of HIV-1 neu tralizing response and for designing AIDS vaccines.
Antibodies, Monoclonal ; genetics ; immunology ; Antibodies, Neutralizing ; genetics ; immunology ; Enzyme-Linked Immunosorbent Assay ; methods ; HEK293 Cells ; HIV Antibodies ; genetics ; immunology ; HIV Envelope Protein gp120 ; genetics ; immunology ; HIV Infections ; immunology ; virology ; HIV-1 ; genetics ; immunology ; isolation & purification ; Humans ; Immunoglobulin Fab Fragments ; genetics ; immunology ; Neutralization Tests ; Peptide Library ; Transfection

Alveolar bone is an important anatomic basis for implant-supported denture restoration, and its different degrees of defects determine the choices of bone augmentation surgeries. Therefore, the reconstruction of alveolar bone defects is an important technology in the clinical practice of implant restoration. However, the final reconstructive effect of bone quality, bone quantity and bone morphology is affected by many factors. Clinicians need to master the standardized diagnosis and treatment principles and methods to improve the treatment effect and achieve the goal of both aesthetic and functional reconstruction of both jaws. Based on the current clinical experience of domestic experts and the relevant academic guidelines of foreign counterparts, this expert consensus systematically and comprehensively summarized the augmentation strategies of alveolar bone defects from two aspects: the classification of alveolar bone defects and the appropriate selection of bone augmentation surgeries. The following consensus are reached: alveolar bone defects can be divided into five types (Ⅰ-0, Ⅰ-Ⅰ, Ⅱ-0, Ⅱ-Ⅰ and Ⅱ-Ⅱ) according to the relationship between alveolar bone defects and the expected position of dental implants. A typeⅠ-0 bone defect is a bone defect on one side of the alveolar bone that does not exceed 50% of the expected implant length, and there is no obvious defect on the other side; guided bone regeneration with simultaneous implant implantation is preferred. Type Ⅰ-Ⅰ bone defects refer to bone defects on both sides of alveolar bone those do not exceed 50% of the expected implant length; the first choice is autologous bone block onlay grafting for bone increments with staged implant placement or transcrestal sinus floor elevation with simultaneous implant implantation. Type Ⅱ-0 bone defects show that the bone defect on one side of alveolar bone exceeds 50% of the expected implant length, and there’s no obvious defect on the other side; autologous bone block onlay grafting (thickness ≤ 4 mm) or alveolar ridge splitting (thickness > 4 mm) is preferred for bone augmentation with staged implant placement. Type Ⅱ-Ⅰ bone defects indicate that the bone plate defect on one side exceeds 50% of the expected implant length and the bone defect on the other side does not exceed 50% of the expected implant length; autologous bone block onlay grafting or tenting techniques is preferred for bone increments with staged implant implantation. Type Ⅱ-Ⅱ bone defects are bone plates on both sides of alveolar bone those exceed 50% of the expected implant length; guided bone regeneration with rigid mesh or maxillary sinus floor elevation or cortical autologous bone tenting is preferred for bone increments with staged implant implantation. This consensus will provide clinical physicians with appropriate augmentation strategies for alveolar bone defects.