OBJECTIVETo observe the content variation of luteolin and luteolin-7-beta-D-glucoside in Chrysanthemum morifolium Ramat. (CMR) from different collection time.

METHODSRP-HPLC was used to analyze these two components in CMR collected in 2001 and 2002.

RESULTThe content of luteolin was significantly lower than that of luteolin-7-beta-D-glucoside. Furthermore, the former showed no marked changes during collection, while the latter did not varied markedly in early collection but decreased significantly in later collection.

CONCLUSIONThe content of luteolin-7-beta-D-glucoside reflects the quality of Chrysanthemum morifolium Ramat. more viably than that of luteolin.

Chromatography, High Pressure Liquid ; Chrysanthemum ; chemistry ; Flavonoids ; analysis ; Glucosides ; analysis ; Luteolin

ngthen cellular immunity, especially Th1-type immune response to HPV11-E7 DNA vaccine in mice.

Near infrared (NIR) spectroscopy as a kind of rapid process analysis technology has been successfully applied in Chinese medicine pharmaceutical process. In this research, the technology was adopted to establish the rapid quantitative analysis models of main indicators from the Lonicera japonica and Artemisia annua alcohol precipitation process of Reduning injection. On-line NIR spectra of 142 samples from alcohol precipitation process were collected and the content of main indicators for each sample were detected through off-line HPLC. With eliminating outliers, determination of spectra pretreatment method and selecting optimal band, the NIR quantitative calibration model for each indicator was established using partial least squares (PLS). These models were used to predict the unknown samples from precipitation process of Reduning injection to achieve the goal of rapid detection. The results showed that the models were ideal. The correlation coefficients of models for neochlorogenic acid, chlorogenic acid, 4-O-caffeoylquinic acid and secoxyloganin were 0.973 872, 0.985 449, 0.975 509 and 0.979 790, respectively and their relative standard errors of prediction (RSEP) were 2.922 49%, 2.341 37%, 2.930 40% and 2.184 60%, respectively. This study indicated that the NIR quantitative calibration model showed good stability and precision, and it can be used in rapid quantitative detection of main indicators of efficacy in order to on-line monitor the alcohol precipitation process of Reduning injection.
Chemical Precipitation ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Ethanol ; chemistry ; Lonicera ; chemistry ; Quality Control ; Spectroscopy, Near-Infrared ; methods

In order to separate and purify the PEGylated recombinant human granulocyte stimulating factor (rhG-CSF) at large laboratory-scale level, a two-step ion-exchange chromatographic separation procedure was designed. Cation-exchange chromatography was applied first to separate PEGylated rhG-CSF from un-reacted rhG-CSF, followed by anion-exchange chromatography to dissolve individual PEG-rhG-CSF species (mono-, di- and tri-PEGylated rhG-CSF) and remove the free PEG. The molecular weight of individual PEGylated rhG-CSF was determined by MALDI-TOF and SDS-PAGE. MALDI-TOF mass spectrometry revealed that the molecular weights of mono-, di- and tri-PEGylated rhG-CSF are 23.8 kD, 28.6kD and 33.8kD, respectively. Cell proliferation activity was detected by MTT assay using NFS-60 cell. The in vitro residual bioactivity of mono-, di- and tri-PEGylated rhG-CSF were 90%, 75% and 43% respectively, comparing with the un-conjugated rhG-CSF. These results indicated that the un-conjugated rhG-CSF and excess free PEG can be removed completely and the three conjugate species can be purified into homogeneity by the two consecutive ion-exchange chromatographic steps. The purification procedure is easy to scale-up, high in performance and recovery.
Chromatography, Ion Exchange ; methods ; Granulocyte Colony-Stimulating Factor ; biosynthesis ; chemistry ; isolation & purification ; Humans ; Polyethylene Glycols ; chemistry ; isolation & purification ; Recombinant Proteins

Objective To evaluate the effects of the anticipated health education plan in patients with prolapse of lumbar intervertebral disc.Methods The anticipated health education plan was established.The patients with window treatment of lumbar disc herniation or lumbar disc herniation and a half laminectomy discectomy were randomly divided into two groups, patients in treatment group were treated with the anticipated health education plan and patients in control group were treated with the conventional health education.The standard-reaching rate of health education and the satisfaction of the patients were investigated at the day before their discharge.And the results were proposed as the evaluation index for estimate of education effect.To compare the health-related quality of life, the SF-36 was completed by the patients before and after surgery at least 6 months.Results In the observed group,the standard-reaching rate of health education was 100.00%,the control group was 74.19%, there were significant differences between the two groups (x2 = 7.032, P ＜0.01); the patient' s satisfaction was 100.00% in the observed group,80.65% in the control group,there were significant differences between the two groups (x2 = 4.613, P ＜ 0.05).The scores of SF-36 were no significant difference between the two groups before surgery (P ＞ 0.05); The total effective rate of observed group was significantly higher than that on control groups with statistical significance (P ＜ 0.05).Conclusions The anticipated health education plan can satisfy the patients, continually improve nursing quality and the life quality of the patients obviously, which is worthy of clinic application.

This study explored the antibiotic resistance and resistance gene carriage of 140 Salmonella strains isolated from environmental sewage in Guangzhou city between March 1,2022,and November 30,2022.The micro broth dilution method was used to select 17 antibiotics for susceptibility testing.According to whole genome sequencing results,the CARD resistance database was used to obtain corresponding resistance genes.High antimicrobial resistance rates above 80％were observed a-gainst ampicillin,tetracycline,streptomycin,chloramphenicol,and cotrimoxazole.The intermediation rate of polymyxin E and ciprofloxacin exceeded 60％.The multiple drug resistance status was severe,and the rate of multiple drug resistance was as high as 92.86％.The strains carried multiple types of resistance genes,particularly for aminoglycosides,with a carriage rate as high as 92.68％.The resistance of Salmonella in environmental wastewater in Guangzhou to one or more drugs was severe,and the overall multi-drug resistance rate gradually increased over time.The resistance spectrum was diverse,and the resistance mechanism,mediated by mobile genetic elements such as re-sistance genes,was found to be the main cause of resistance to one or more drugs.

OBJECTIVETo detect the cancer stem cells and to evaluate their prognostic implication in patients with lung adenocarcinoma.

METHODSThree phenotypic markers of cancer stem cells (SP-C, CCSP and OCT4) in lung adenocarcinoma were detected by immunofluorecence staining. The correlation among the clinicopathological parameters and phenotypes of cancer stem cells as well as survival were analyzed by Cox proportional hazard method.

RESULTSOf the 57 cases, cancer stem cells were detected in 52, including OCT4(+) bronchioloalveolar stem cell (BASC) phenotype (SP-C(+) CCSP(+) OCT4(+)) in 40 cases and OCT4(-) BASC phenotype (SP-C(+) CCSP(+) OCT4(-)) in 12 cases. Statistical analysis revealed that the phenotype of cancer stem cells was related with the cellular differentiation, i.e. the OCT4(+) BASC phenotype occurred more frequently in the well-differentiated tumors, while the OCT4(-) BASC phenotype usually presented in most of the poorly-differentiated ones. Cox analysis showed that the OCT4(+) BASC phenotype was one of prognostic factors.

CONCLUSIONThe lung adenocarcinoma stem cells have phenotypic features of bronchioalveolar stem cells (SP-C(+) CCSP(+)). The expression of self-renewal regulatory gene OCT4 in these cells indicates an aggressive nature and unfavorable prognosis.

Adenocarcinoma ; genetics ; metabolism ; pathology ; Adult ; Aged ; Aged, 80 and over ; Cell Differentiation ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Neoplastic Stem Cells ; metabolism ; pathology ; Octamer Transcription Factor-3 ; genetics ; metabolism ; Phenotype ; Proportional Hazards Models ; Pulmonary Surfactant-Associated Protein C ; genetics ; metabolism ; Survival Rate ; Uteroglobin ; genetics ; metabolism

OBJECTIVEThis study aimed at exploring the feasibility of using dried blood spots (DBS) to detect HIV drug resistance genotyping in China by comparing the results of drug resistance from DBS, plasma and whole blood samples.

METHODSBlood samples were collected from 39 AIDS patients from Anhui (10), Yunnan (13), Hunan (6) and Xinjiang (10) provinces and autonomous regions. The HIV strains that infected these patients covered all the major HIV-1 subtypes prevailing in China (B, CRF01_AE, CRF07_BC). HIV drug resistance genotyping assay was performed on DBS as well as on the whole blood and plasma samples from the same patients simultaneously by using an in-house nest RT-PCR method. Drug resistance levels were determined based on Stanford University HIV drug resistance database, and the results from these three types of samples were compared.

RESULTSThe percentages of successful amplification of protease and reverse transcriptase regions in the pol gene were 95% (37/39) from DBS, 92% (36/39) from whole blood and 100% (39/39) from plasma samples. The sequences from the three types of samples showed more than 99% identity.86% (31/36) of the DBS samples had the same set of drug resistance mutations as those which were detected from plasma samples. The differences probably resulted from mixed bases.

CONCLUSIONSThere was no major difference in detecting HIV drug resistance genotyping among DBS, plasma and whole blood samples. Therefore, DBS is useful for detection of HIV drug resistance genotyping and is particularly valuable in developing countries like China, especially in remote rural regions.

Dried Blood Spot Testing ; Drug Resistance, Viral ; genetics ; Feasibility Studies ; Genotype ; HIV Infections ; blood ; genetics ; virology ; HIV Seropositivity ; blood ; genetics ; virology ; HIV-1 ; drug effects ; genetics ; Humans ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Load

Objectives: To observe the effect of activated G-protein coupled estrogen receptor 1 (GPER1) on Angiotensin II (AngII)-induced hypertrophy of cultured neonatal rat cardiomyocytes and explore related mechanisms. Methods: Primary cardiomyocytes derived from 2-to 3-day-old neonatal rats were cultured in vitro. Tandem mass tags (TMT) protein mass spectrometry was used to examine protein expressions; relevant bioinformatics analysis was performed to screen the possible regulatory mechanisms. Cardiomyocytes were divided into 6 groups: (1)Blank control group, (2) AngII group, (3)AngII+G1 (GPER1 activator) group, (4)AngII+G1+G15 (GPER1 inhibitor) group, (5)AngII+G1+U0126 (extracellular ERK inhibitor) group and (6)AngII+G1+MK2206 (AKT inhibitor) group (n=3 for each group). Cardiomyocytes GPER1 expressions, mRNA levels of atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP), protein levels of ERK, AKT with their interactions, autophagy-related proteins LC3II and LC3I were compared among different groups;impact of GPER1 on cardiomyocytes apoptosis was detected by flowcytometry. Results: AngII induced cardiomyocytes hypertrophy and upregulation of ANP and BNP mRNA levels in a dose-dependent manner (P<0.05). GPER1 expression could be detected on cardiomyocytes by Immunofluorescence technique. qRT-PCR results showed that GPER1 was activated by the specific activator G1 and mRNA expressions of ANP and BNP were inhibited in a dose-dependent manner by the specific activator G1 (P<0.05); mRNA levels of ANP and BNP were re-elevated in AngII+G1+G15 group (P<0.05). Western blotting results showed that protein expression of p-ERK and p-AKT was significantly higher in AngII group and AngII+G1 group than in blank control group (P<0.05), significantly reduced in AngII+G1+G15 group compared with AngII+G1 group (P<0.05); decreased expressions of p-ERK, p-AKT and mRNA levels of ANP,BNP were also detected in AngII+G1+MK2206 group (P<0.05). G1 induced protein expression was similar between AngII+G1 group and AngII+G1+U0126 group. Flowcytometry results indicated that cardiomyocytes apoptosis was similar between AngII+G1 group and AngII group (P>0.05). Ratio of LC3II/LC3I was significantly higher and autophagy levels were significantly enhanced in AngII group than in blank control group (P<0.01), these changes could be significantly reversed in AngII +G1 group (P<0.01 vs. AngII). Conclusions: Activation of GPER1 could inhibit neonatal cardiomyocytes hypertrophy, this effect might be related to AKT and ERK signaling pathway and cell autophagy.

Objective To observe the clinical efficacy of acupuncture plus intradermal needle in treating mild-to-moderate post-stroke depression (PSD). Method Ninty patients with mild-to-moderate PSD were randomized into group A, B and C, with 30 cases in each group. All the groups were intervened by selecting Baihui (GV20), Shenting (GV24) and Yintang (GV29). Group A received acupuncture plus intradermal needle; group B, intradermal needle alone; group C, acupuncture plus sham acupuncture. Hamilton depression rating scale (HDRS) score and Barthel index (BI) score in the three groups were observed before and at the end of the treatment, and the clinical efficacies of the three groups were compared. Result The HDRS score and BI score showed a significant change after the treatment in the three groups (P<0.01). After the treatment, the HDRS score and BI score in the group A and group B was significantly different from those in group C (P<0.05). The total effective rate was 90.0％ in group A versus 86.7％in group B, and 73.3％ in group C; there was a statistically significant difference between group C and group A or B (P<0.01, P<0.05). Conclusion Both acupuncture plus intradermal needle and intradermal needle alone are effective approaches in treating mild-to-moderate PSD. The treatment efficacy of these two methods is similar.