This study was conducted to investigate the paclitaxel loaded by hydrazone bonds in poly(ethylene glycol)-poly(caprolactone) micelles (mPEG-PCL-PTX) on proliferation and apoptosis of human lung cancer A549 cells and its possible mechanisms of anti-tumor activity. The cell proliferation was measured with MTT assay. Flow cytometry were used to analyze the cell cycle. The cell apoptosis was analyzed using Hoechst/P staining. The expression levels of apoptotic genes expression in the mitochondrial apoptosis pathway were detected by RT-PCR and Western blotting, respectively. The mPEG-PCL-PTX could inhibit the proliferation of A549 cells and promote the apoptosis. The Bax, caspase-3 protein expression were increased while Bcl-2 protein expression was decreased in A549 cells. Results showed that the polymer containing hydrazone bond is non-toxic in vitro, the mPEG-PCL-PTX micelles can inhibit the proliferation and induce the apoptosis of A549 cells. Key words: paclitaxel; micelle; A549 cell; proliferation; cell cycle; apoptosis
Apoptosis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Micelles ; Paclitaxel ; pharmacology ; Polyesters ; Polyethylene Glycols ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

The conversion of exogenous p-hydroxybenzaldehyde to p-hydroxy-methyl-phenol-beta-D-glucoside (gastrodin) was studied by using cell suspension culture of Datura tatula L. The chemical structure of this synthesized gastrodin was identified based on the spectral analysis and chemical evidence. The conversion procedure of p-hydroxybenzaldehyde into gastrodin by D. tatula L. cell suspension cultures was established. The synthesized gastrodin (II) was isolated from the ferment liquor and identified by spectral analysis. At the same time, the p-hydroxybenzyl alcohol (I) converted through biotransformation of p-hydroxybenzaldehyde by cell suspension cultures of D. tatula L. was also isolated and identified. The efficiency of glucosylation of p-hydroxybenzaldehyde was remarkably enhanced by adding salicylic acid (0.1 mg/L) and keeping the lower pressure (0.001MPa) in 25L airlift loop bioreactor. The biotransformation of exogenous p-hydroxybenzaldehyde to gastrodin by cell suspension culture of D. tatula L. is a promising approach.
Benzaldehydes ; metabolism ; Benzyl Alcohols ; chemistry ; Bioreactors ; Biotransformation ; Datura ; metabolism ; Glucosides ; biosynthesis ; chemistry ; Salicylic Acid ; pharmacology ; Suspensions

AIMTo investigate the production of p-hydroxymethylphenol-beta-D-glucoside (gastrodin) through biotransformation by plant cell suspension cultures.

METHODSUsing cell suspension cultures of Datura stramonium to convert the exogenous p-hydroxybenzaldehyde into gastrodin was conducted and the converted compounds were separated with a combination of multi-chromatography. Their chemical structures were determined on the basis of spectral analysis and chemical evidence.

RESULTSThe conversion procedure of p-hydroxybenzaldehyde into gastrodin by Datura stramonium cell suspension cultures was established. The synthesized gastrodin (II) was isolated from the fermental liquor and identified by spectral analysis. At the same time, the p-hydroxybenzyl alcohol (I) converted through biotransformation of p-hydroxybenzaldehyde by cell suspension cultures of Datura stramonium was also isolated and identified. Two compounds were also isolated from the cell cultures and they were identified as beta-D-furanoallulose (III) and n-butyloxystyryl-beta-D-pyranoallulose (IV).

CONCLUSIONDatura stramonium grown in suspension cultures can convert exogenous p-hydroxybenzaldehyde into the corresponding gastrodin.

Benzaldehydes ; metabolism ; Benzyl Alcohols ; isolation & purification ; metabolism ; Biotransformation ; Cell Culture Techniques ; methods ; Cells, Cultured ; Datura stramonium ; cytology ; metabolism ; Glucosides ; biosynthesis ; isolation & purification ; Plant Stems ; cytology ; Plants, Medicinal ; cytology ; metabolism

Aim To observe the effect of ilexonin A (IA) on the proliferation of bone marrow mesenchymal stem cells(BMSCs),and to investigate whether IA can promote the migration of BMSCs by up-regulating the expression of CXCR4 in rats. Methods MTT method was used to assay and analyse the proliferation of BM-SCs which were pretreated with different concentrations of IA (3.125,6.25,12.5,25,50,100,200,400, 800 mg·L-1) for 24,48 and 72h,then the best con-centration and the best optimum time were screened. The third generation of BMSCs was exposed to the opti-mal concentration of IA for 48h. The Transwell system was used to carry out the experiment of BMSCs migra-tion. Western blot was used to analyse the expression of CXCR4. Results MTT assay showed that com-pared with control group, the proliferation of BMSCs was significantly reduced in IA 100 ~800 mg·L-1 groups at 24h(P < 0.05); compared with control group, the proliferation of BMSCs significantly de-creased in IA 100~800 mg·L-1groups at 48h(P<0.05),but markedly increased in IA 6.25 and 3.125 mg·L-1groups (P <0.05); compared with control group,the proliferation of BMSCs was significantly re-duced in IA 12.5~800 mg·L-1groups at 72h(P<0.05). The above results indicated that the BMSCs in-cubated with IA 6.25 and 3.125 mg·L-1for 48h were the optimal choice to promote proliferation. The Transwell migration assay showed that incubation with IA 6.25 and 3.125 mg·L-1for 48h could significant-ly increase the migration of BMSCs(P <0.05), and the migration rate was not related with the concentra-tion of IA. This effect was completely blocked by AMD3100(the antagonist of CXCR4). Western blot showed that incubation with IA 6.25 and 3.125 mg· L-1for 48h could increase the expression of CXCR4 in BMSCs(P<0.05). Conclusion IA can promote the proliferation of BMSCs and increase the migration of BMSCs by up-regulating the expression of CXCR4.

Objective To investigate Notch1 protein expression of leukemic cells in children with pediatric Bcell acute lymphoblastic leukemia(B-ALL) and its effect on prognosis.Methods The expression of Notch1 protein of leukemic cells in bone marrow smears was detected by immunohistochemistry(SABC) in primarily diagnosed childhood ALL,including 63 cases of B-ALL and 14 cases of T-ALL,a group of 34 children with no malignancy served as controls.Reverse transcription-ploymerase chain reaction was used to assay Notch1 mRNA expression in ALL patients.Results The incidence of Notch1 expression was 31.7％ in B-ALL patients,71.4％ in T-ALL patients,and 8.8％ in control group,respectively.Notch1 protein was aberrantly expressed in both B-ALL and T-ALL compared with the controls(P ＜ 0.05,P ＜ 0.001).Multivariate analysis revealed that the expression of Notch1 protein in B-ALL was not associated with patients' age,gender,WBC count at diagnosis(all P ＞ 0.05),it had no influence on the early treatment response.Nevertheless,the Kaplan-Meier curve of event-free survival showed that,in the patients without Notch1 protein expression,the long-term event-free survival rate was as high as 92.7 ％,in contrast,in the children with Notch 1 expression,the event-free survival was only 54.5 ％ (P =0.0054).Conclusions The expression of Notch 1 protein in pediatric B-ALL may predict a poor prognosis,and interfering with Notch1 signaling could be employed as a potential therapeutic target for those patients with Notch1 expression.

Objective:To investigate the effects of self-made Gujin Xiaoji Mixture combined with warming needle therapy on the clinical efficacy and immune function of patients with advanced non-small cell lung cancer (NSCLC) with qi and yin deficiency syndrome.Methods:This experiment was a randomized controlled trial study. 180 patients with advanced NSCLC qi and yin deficiency syndrome in the oncology centre of Qingdao Hospital of Traditional Chinese Medicine were selected as the observation subjects from March 2021 to August 2022, and were divided into 2 groups using the random number table method, with 90 cases in each group. The control group received conventional chemotherapy combined with Sintilimab injection, 21 days as a cycle, with a total of 4 cycles of treatment; and the observation group received Gujin Xiaoji Mixture combined with warming needle therapy based on the control group, 7 days as one course of treatment, with a total of 12 courses. Both groups were followed up for 12 months. The TCM syndrome scores were performed before and after treatment. The functional assessment of cancer therapy-lung (FACT-L) was used to evaluate the quality of life of patients; flow cytometry was used to detect the levels of CD3 +, CD4 +, CD8 + and NK cell, and the CD4 +/CD8 + ratio was calculated; adverse drug reactions and progression free survival of patients during treatment were observed and recorded, the efficacy of TCM syndrome and objective efficacy of solid tumors were evaluated. Results:After treatment, the observation group's post-treatment TCM syndrome score (5.67±1.99 vs. 7.12±2.31, t=-4.53) was lower than that of the control group ( P<0.001); mobility (23.03±2.80 vs. 20.69±2.46, t=5.96), daily living (23.06±2.56 vs. 20.71± 2.33, t=6.42), emotional status (18.44±2.32 vs. 16.12±2.71, t=6.18), and other factors (33.14±4.11 vs. 27.39±4.64, t=8.81) and total score (97.68±7.23 vs. 84.91±7.49, t=11.64) were higher than those in the control group ( P<0.01). In the observation group, after treatment, the levels of CD3 + [(65.14±6.06)% vs. (59.84±5.74)%, t=6.02], CD4 + [(40.09±4.09)% vs. (35.69±3.86)%, t=7.43], NK cell [(29.11±4.81)% vs. (22.38±4.51)%, t=9.68] and CD4 +/CD8 + [(1.52±0.27) vs. (1.14±0.12), t=12.63] were higher than those in the control group ( P<0.01), and CD8 + [(26.82±3.79)% vs. (31.76±4.65)%, t=-7.81] level was lower than that of the control group ( P<0.01). After treatment, the objective remission rate in the observation group was 7.8% (7/90), and the disease control rate was 87.8% (79/90), while the objective remission rate after treatment in the control group was 5.5% (5/90), and the disease control rate was 82.2% (74/90), and there were no statistical significance in the comparison of objective remission rate and disease control rate of the 2 groups ( χ2=0.09, 0.70, P=0.765, 0.407). The total effective rate after treatment was 62.2% (56/90) in the observation group and 34.4% (31/90) in the control group, and the difference between the 2 groups was statistically significant ( Z=-3.89, P<0.001). WBC [(4.27±1.12)×10 9/L vs. (3.84±1.11)×10 9/L, t=2.58] and haemoglobin [(119.93±17.25)g/L vs. (109.76±15.61)g/L, t=4.15] levels of the observation group were higher than those in the control group after treatment ( P<0.01). During follow-up, the median progression-free survival was 6.2 months in the observation group and 5.5 months in the control group patients, and the difference between the 2 groups was not statistically different ( t=0.11, P>0.05). Conclusion:The combination of Gujin Xiaoji Mixture with warming needle therapy can effectively improve the clinical symptoms of patients with advanced NSCLC with deficiency of qi and yin syndrome, improve the immunity and clinical efficacy of patients, alleviate the adverse effects of drugs, and prolong the progression-free survival period.

OBJECTIVETo investigate the clinicopathologic characteristics of childhood renal diseases.

METHODSA retrospective analysis of 1316 renal biopsies performed over the past 20 years was performed.

RESULTSOf the 1316 patients, 383 (29.09% ) were diagnosed as nephrotic syndrome, 291 (22.00%) as acute nephritis syndrome, 224 (17.21%) as isolated hematuria, 209(15.87%) as purpura nephritis, and 96 (7.30% ) as hepatitis B virus-associated nephritis . Mesangial proliferation was the most common pathological change (756 cases; 57.45%), followed by IgA nephropathy (113 cases; 8.59%), endothelial capillary proliferation(112 cases; 8.51%), membranous nephropathy (66 cases; 5.02%), and various minor and minimal changes (59 cases; 4.48%). Alport syndrome, congenital nephrotic syndrome, thin basement membrane nephropathy, fibrillary glomerulopathy disease, and Fabry disease were confirmed by electronic microscopy. IgA, IgM and C1q nephropathy were definitely diagnosed using immune histochemistry or immunofluorescent. A diagnosis of primary glomerular disease was made in 69.53% of the cases (915 cases); secondary glomerular disease was noted in 26.14% (344 cases). Of the 915 cases of primary glomerular disease, 375 (41.0%) had nephrotic syndrome. Secondary glomerular disease due to purura nephritis was common (209/344; 60.8%).

CONCLUSIONSPrimiary glomerular disease predominates in children. Nephrotic syndrome is the most common clinical diagnosis. Mesangial proliferation is the most common pathological patterns in children with renal disease.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Kidney ; pathology ; Kidney Diseases ; pathology ; Kidney Glomerulus ; pathology ; Male ; Renal Insufficiency ; pathology ; Retrospective Studies

OBJECTIVEThis clinical study was designed to evaluate the efficacy and toxicity of the combined regimen of docetaxel, 5-Fu and DDP (TPF) in the treatment of advanced or relapsed nasopharyngeal carcinoma (NPC).

METHODSFifty-six patients with newly diagnosed or recurrent/metastatic NPC following chemotherapy or radiotherapy were enrolled. Both docetaxel and DDP were administered intravenously for 6 hours at the dose of 70 mg/m2 on D1. 5-Fu was given at a dose of 400-500 mg/m2 for 6 hours from D1 to D5. Dexamethasone was routinely administered before injection of docetaxel. This combination was repeated every 3 to 4 weeks, and continued for 4-6 cycles or until PD for the responders.

RESULTSFifty-one (91.1%) patients were evaluable for response assessment. The response rate for whole group was 72.5% (37/51) with a CR rate of 9.8% (5/51). The stable disease accounted for 17.6% (9/51). There were 17(30.4%) chemotherapy-naïve patients. The overall response rate in those was 82.4% with a CR rate of 29.4%. However, the response rate for previously treated patients was 64.7% without CR. Twelve patients had progressed disease, including 5 (8.9%) died of disease progression with a median follow-up of 11 month (ranged from 1 to 19 months). Totally, 196 courses of chemotherapy were administered. The major toxicity was myelosupression, nausea/vomiting. The incidence of leucopenia was 48% with 22.2% of these in NCI grade II or IV. But only 2 patients (3.6%) experienced leucopenia with a fever. Other mild toxicities including alopecia, asthenia, mucositis and diarrhea were also observed.

CONCLUSIONOur preliminary outcome shows docetaxel, 5-Fu and DDP combination is effective and safe for the patients with advanced or relapsed nasopharyngeal carcinoma. But further clinical study is warranted.

Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma, Squamous Cell ; drug therapy ; pathology ; Cisplatin ; administration & dosage ; Female ; Fluorouracil ; administration & dosage ; Follow-Up Studies ; Humans ; Leukopenia ; chemically induced ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; drug therapy ; pathology ; Nausea ; chemically induced ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Remission Induction ; Taxoids ; administration & dosage ; Young Adult

To investigate the existence of tick-borne pathogens infection of rodents at the border of China and the Demo-cratic People's Republic of Korea(DPRK).PCR was used to detect the spotted fever group rickettsiae(SFGR)ompA gene,Ehrlichia chaffeensis(Ec)and Anaplasma phagocytophilum(Ap)16S rRNA,Candidatus Neoehrlichia mikurensis(CNm)groEL gene,Bartonella(Ba)rpoB gene,and Francisella tularensis(Ft)fopA gene in rodents samples collected from Ji'an of Jilin province and Kuandian of Liaoning Province.The positivity rates of 132 wild rats spleen samples,SFGR,Ec,Ap,CNm,Ba,and Ft were 9.85％,12.88％,5.30％,3.79％,51.52％,and 6.06％,respectively,with statistical differences in in-fection rates(x2=149.236,P=0.000).The infection rate of Ba was the highest in wild rats in this area.There was no signifi-cant difference in the infection rate of SFGR,Ec,Ap,CNm,and Ft among different rats species,but there were significant differences in the infection rate of Ba(x2=13.36,P=0.010).The infection rate of Apodemus agrarius was the highest.A-mong 132 wild rats specimens,the coinfection rate of the two pathogens was 15.9％(21/132),with Ba as the main species(15/132),and two cases of coinfection with three pathogens were detected.The infection of six tick-borne pathogens is common in wild rats at the China/DPRK border.Co-infection of two or three pathogens indicates a risk of multiple tick-borne pathogens and mixed natural foci of multiple tick-borne infec-tious diseases.

Aim To investigate the protective effect of 1, 8-Cineole on the injury of human aortic endothelial cells (HAECs) induced by high glucose (HG) via regulating autophagy. Methods Cells were incubated with different doses of 1, 8-Cineole followed by exposing to HG for 60 h, and MTT assay was used to analyse cell viability, lactate dehydrogenase (LDH) was used to detect cytotoxicity, and Western blot was used to detect Beclin1, LC3-II/I, p62, caspase-3 and caspase-9 expressions. Autophagy inhibitor (chloroquine, CQ) was treated on HAECs, and the expressions of Beclinl, LC3-II/I, p62, caspase-3 and caspase-9 were measured by Western blot. Results 1, 8-Cineole increased cell viability, reduced the content of LDH, activated autophagy and inhibited apoptosis. Compared with control group, the expression of Beclinl, LC3-II/I, p62, caspase-3 and caspase-9 in CQ group increased. Simultaneously, the expression of above-mentioned between CQ + HG group and CQ + HG + CH group. Conclusions 1, 8-Cineole has protective effect on the injury of HAECs induced by high glucose, and its effect is related to improving autophagy flux.