AIM: To establish a method for the quality standard of Chanfukang Granules (Radix Astragali, Herba Leonuri, Fructus Aurantii, Herba Agrimoniae, Radix Rehmanniae, etc.). METHODS: TLC was used for identification of Herba Leonuri, Fructus Aurantii, Herba Agrimoniae, Radix Rehmanniae. The content of astragaloside Ⅳ was determined by HPLC-ELSD. RESULTS: The TLC identification was highly specific and the spots clear and concentrated. The linear range of astragaloside Ⅳ was 0.336-2.016 ?g, r=0.999 4. The average recovery was 97.13% and RSD was 1.3%. CONCLUSION: The method is simple and accurate. It can be used for quality control of Chanfukang Granules.

Adult ; Age Distribution ; Aged ; Aged, 80 and over ; DNA, Viral ; blood ; Evaluation Studies as Topic ; Female ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; isolation & purification ; Hepatitis B, Chronic ; blood ; virology ; Humans ; Liver Cirrhosis ; blood ; virology ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Reagent Kits, Diagnostic ; Sensitivity and Specificity ; Viral Load

OBJECTIVETo study the effect of human spastic paraplegia 21 protein (SPG21) on the replication of hepatitis B virus(HBV) and its regulatory mechanism.

METHODSHBV infectious clone pHBV1.3 and its promoter pHBV-Luc were transfected respectively into HepG2 cells with SPG21 of different concentrations, HBsAg and HBeAg in the supernatants were measured by enzyme linked immunosorbent assay (ELISA), expression of HBV core mRNA and protein were detected by RT-PCR and western blot, covalently closed circular DNA(ccc DNA) levels were measured by real-time PCR, and HBV promoter activity was measured by luminometer fluorescence detector.

RESULTSExpression of HBsAg, HBeAg, HBV core protein and cccDNA were upregulated by SPG21 as well as HBV promoter activity in a dose-dependent approach. The activity of HBV promoter increased to 1.63, 3.09 and 4.66 times in HepG2 cells treated with 50mug/ml, 100mug/ml and 200mug/ml SPG21 respectively during 48 hour-treated ( P less than 0.05), as compared to the control group.

CONCLUSIONSSPG21 can enhance the replication of HBV in HepG2 cells.

Adaptor Proteins, Signal Transducing ; metabolism ; Hep G2 Cells ; Hepatitis B virus ; metabolism ; physiology ; Humans ; Transfection ; Virus Replication

Adolescent ; Adult ; Aged ; DNA, Viral ; Female ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; genetics ; Humans ; Male ; Middle Aged ; Mutation ; Young Adult

OBJECTIVETo investigate if an adenovirus vector carrying antisense multidrug resistance gene 1 (MDR1) could reverse multidrug resistance (MDR) of HepG2/ adriamycin (ADM) cells in tumors transplanted in athymic mice.

METHODSAn adenovirus vector carrying AFP promoter and antisense MDR1 was constructed. HepG2 MDR cells (HepG2/ADM) were induced by graded resistance to ADM and were subcutaneously inoculated into athymic mice to construct the transplanted tumor. After adeno-asmdr1 was injected, the volume of the transplanted tumor and the apoptotic body in the xenograft tumor cells were observed and reverse transcriptase polymerase chain reaction was employed to investigate the expression of the mdr1-mRNA from the mouse transplanted tumor cells.

RESULTSFollowing injection with adeno-asmdr1, the tumor volumes in this mice group did not increase. However the tumor volume in the PBS plus ADM group did increase significantly (P less than 0.05). In the tumor xenograft cells, mdr1 mRNA in the xenografts was assessed by RT-PCR and found to be reduced at week 1, and at week 4 in the ADM+asmdr1 group, but it was stable in the ADM group. It was only 20% in the ADM+asmdr1 group compared to the ADM group at the 4th week. Evidence of apoptosis was observed in the tumor xenograft cells treated with adeno-asmdr1, but there was rarely any apoptosis in the group treated with ADM and PBS.

CONCLUSIONAdenovirus carrying antisense mdr1 RNA can partially reverse the MDR of HepG2/ADM cells and inhibit tumor growth by down-regulating mdr1 mRNA resulting in tumor cell apoptosis.

ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Adenoviridae ; genetics ; Animals ; Carcinoma, Hepatocellular ; drug therapy ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; drug effects ; genetics ; Genetic Vectors ; Hep G2 Cells ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; RNA, Antisense ; genetics

Animals ; Antigen-Antibody Complex ; analysis ; DNA ; immunology ; DNA, Protozoan ; immunology ; Female ; In Situ Hybridization ; Lupus Nephritis ; etiology ; Mice ; Mice, Inbred BALB C ; Trypanosoma ; genetics

Objective To explore the predictive value of serum homo sapiens microRNA(hsa-miR)-30c-5p expression in patients with type 2 diabetes mellitus(T2DM)for microvascular complications.Methods A total of 205 T2DM patients admitted to Bazhong Central Hospital from May 2021 to September 2022 were selected as the diabetes group,and the diabetes group was further divided into diabetes with combined group(n=124)and non combined group(n=81)according to the microvascular complications of the patients.In addition,205 healthy people who underwent physical examination during the same period were selected as the control group.The expression of hsa-miR-30c-5p in serum was detected by reverse transcription-polymerase chain reaction(RT-PCR)and compared.The factors affecting microvascular complications were analyzed using multivariate logistic regression analysis,and receiver operating characteristic(ROC)curves were plotted to predict the value of serum hsa-miR-30c-5p expression in predicting microvascular complications in T2DM patients.Results The expression of serum hsa-miR-30c-5p in the combined group(0.58±0.06)and the non-combined group(0.72±0.08)were lower than that in the control group(0.89±0.21),and the differences were significant(t=16.038,7.079,all P=0.001).The combined group was lower than the non-combined group,and the difference was significant(t=14.289,P=0.001).The course of diabetes[(OR(95％CI):3.873(2.976～4.770)],uric acid[(OR(95％CI):2.125(1.211～3.040)]and glycosylated hemoglobin[(OR(95％CI):2.680(1.745～3.616)]were independent risk factors for microvascular complications in T2DM patients(all P＜0.05),while the time within the target range of glucose[(OR(95％CI):0.491(0.135～0.846)]and serum hsa-miR-30c-5p[(OR(95％CI):0.532(2.976～4.770)]were protective factors for microvascular complications in T2DM patients(all P＜0.05).The sensitivity,specificity and area under the curve(95％CI)of serum hsa-mir-30c-5p expression in predicting microvascular complications in T2DM patients were 81.45％,85.19％and 0.802(0.741～0.854),respectively.Conclusion The expression of serum hsa-miR-30c-5p in patients with T2DM is abnormally reduced,and serum hsa-miR-30c-5p is a protective factor for microvascular complications in patients with T2DM.It may have a certain predictive value for microvascular complications in patients with T2DM.

OBJECTIVETo isolate and characterize human rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLSs).

METHODSThe synovial membrane tissues were obtained from 4 RA patients, 1 chondroma patient and 1 healthy subject and FLS were isolated by means of tissue culture. The cell morphology was observed by phase-contrast microscope and the cell surface markers were detected by flow cytometry.

RESULTSThe FLSs were successfully cultured from the synovial membrane tissues with good cell homogeneity after the third passage. The FLSs of the 3rd to 7th passages were stable and proliferated actively, followed by slow proliferation and aging since the 8th passage. Flow cytometry showed that the 4th-passage FLSs from the RA patients contained 99.04% CD90(+) cells, 2.73% CD3(+) cells, 0.29% CD3(-)CD19(+) cells, 2.81% CD3(-)CD16(+)CD56(+) cells, 5.89% CD14(+) cells, and 54.17% CD55(+) cells. The presence of interleukin-1 receptor type I (IL-1RI, 158.63-/+20.32 pg/ml) and IL-1beta (4.67-/+0.82 pg/ml) were detected in the cell culture supernatant of the 4th-passage FLSs from the RA patients by enzyme-linked immunosorbent assay ELISA.

CONCLUSIONFLSs from RA patients can be effectively culture by means of tissue culture, and the cultured FLSs show high expressions of CD90, IL-1RI and IL-1beta.

Adult ; Aged ; Arthritis, Rheumatoid ; pathology ; Cell Proliferation ; Cell Separation ; Cells, Cultured ; Female ; Fibroblasts ; pathology ; Humans ; Interleukin-1beta ; metabolism ; Male ; Middle Aged ; Receptors, Interleukin-1 Type I ; metabolism ; Synovial Membrane ; cytology ; pathology ; Thy-1 Antigens ; metabolism

OBJECTIVETo investigate the clinical value of different magnetic resonance (MR) pulse sequences in diagnosis of spinal metastatic tumor.

METHODSFifteen patients with clinically suspected spinal metastatic tumor were included in this study. These patients were with documented primary tumors. Four MR pulse sequences, T1-weighted spin echo (T1WI SE), T2-weighted fast spin echo (T2WI FSE), short time inversion recovery (STIR), and gradient echo 2-D multi echo data imaging combination (GE Me-2D) were used to detect spinal metastasis.

RESULTSFifteen vertebral bodies were entire involvement, 38 vertebral bodies were section involvement, and totally 53 vertebral bodies were involved. There were 19 focal infections in pedicle of vertebral arch, 15 metastases in spinous process and transverse process. Fifty-three vertebral bodies were abnormal in T1 WI SE and GE Me-2D, 35 vertebral bodies were found abnormal in T2WI FSE, and 50 vertebral bodies were found abnormal in STIR. The verges of focal signal of involved vertebral bodies were comparatively clear in T1WI SE, comparatively clear or vague in T2WI FSE, vague in STIR, and clear in GE Me-2D.

CONCLUSIONSGE Me-2D may be the most sensitive technique to detect metastases. So three sequences (T1WI SE, T2WI FSE, GE Me-2D) can demonstrate the early changes of spinal metastasis roundly.

Cervical Vertebrae ; diagnostic imaging ; Coccyx ; diagnostic imaging ; Humans ; Lumbar Vertebrae ; diagnostic imaging ; Magnetic Resonance Imaging ; methods ; Neoplasm Metastasis ; pathology ; Radiography ; Sacrum ; diagnostic imaging ; Sensitivity and Specificity ; Spinal Neoplasms ; pathology ; secondary ; Spine ; diagnostic imaging ; Thoracic Vertebrae ; diagnostic imaging

Objective To study the characteristics of micro environmental flow and the process of macromolecular drug delivery in solid tumor. Method3D models simulating tumor hemodynamics and drug delivery were developed. Microcirculation and concentration distribution of two macromolecular drugs in tumor were simulated. ResultsThe model successfully reflected abnormal tumor hemodynamics, which results in impeded drug delivery and uneven concentration distribution of the drug. The results also shows that the drug with lower molecular weight is more easy to spread in tumor and excrete with interstitial fluid flow. In contrast, the drug with higher molecular weight tends to accumulate and maintain an effective concentration in tumor for a longer period. ConclusionsThe results derive from this model can reflect physiological conditions, providing a reliable numerical model for the investigation of tumor micro environment, drug delivery and strategy of therapy.