OBJECTIVE To investigate the mechanism of Xiaoai Jiedu Recipe in promoting ferroptosis of tumor cells and inhibi-ting the growth of transplanted tumors in hepatocellular carcinoma mice by proteomics method.METHODS C57BL/6J mice were randomly divided into,model group,low-and high-dose groups of Xiaoai Jiedu Recipe,Xiaoai Jiedu Recipe combined with ferropto-sis inhibitor group,ferroptosis inhibitor group and cisplatin group.The H22 mouse transplantation tumor model was constructed and the drug administration interventions were as follows:the low-and high-dose groups of Xiaoai Jiedu Recipe were given Xiaoai Jiedu Reci-pe by gavage at the doses of 10 and 20 g·kg-1·d-1;the ferroptosis inhibitor group was given Liproxstatin-1 by intraperitoneal injec-tion at the dose of 10 mg·kg-1·d-1;the cisplatin group was given cisplatin by intraperitoneal injection at the dose of 10 mg·kg-1·d-1;the Xiaoai Jiedu Recipe combined with ferroptosis inhibitor group was given a gavage dose of 20 g·kg-1·d-1of Xiaoai Jiedu Recipe and an intraperitoneal injection of 10 mg·kg-1·d-1 of the ferroptosis inhibitor Liproxstatin-1;the model group was given an equal amount of saline by gavage.The drugs were administered for 11 d continuously.The tumors were stripped to calcu-late tumor inhibition rate.Pathological changes were observed by hematoxylin-eosin(HE).Mitochondrial structural changes were ob-served by transmission electron microscopy.Reactive oxygen species(ROS)levels were detected by flow cytometry.Serum was pre-pared and analysed by TMT peptide labelling combined with LC-MS/MS to find differential protein expression profiles,applying IPA software.Serum iron ions,glutathione(GSH)and malondialdehyde(MDA)levels were measured biochemically.Protein expression levels of nuclear factor E2-related factor 2(Nrf2),heme oxygenase-1(HMOX1),cystine/glutamate reverse transporter protein(SLC7A11)and glutathione peroxidase 4(GPX4)were measured by protein Western blot.RESULTS The tumor growth was inhibi-ted in the low-and high-dose groups and the cisplatin group,with inhibition rates of 36.12%,51.63%and 57.43%,respectively,while the tumor growth was promoted in the ferroptosis inhibitor group,with an inhibition rate of-45.56%,and the tumor size was re-duced in the Xiaoai Jiedu Recipe combined with ferroptosis inhibitor group compared with the ferroptosis inhibitor group,with an inhi-bition rate of 18.11%.HE staining showed that apoptotic cells and a large number of vacuoles accumulated in the tumor tissues of the high-dose group and the cisplatin group.Transmission electron microscopy showed that the mitochondrial atrophy and membrane densi-ty increased to a certain extent in the low and high-dose groups of Xiaoai Jiedu Recipe.Flow cytometry results showed that ROS levels were significantly increased after the intervention of Xiaoai Jiedu Recipe(P<0.01).Proteomic tests showed that there were 129 differ-ential proteins,including 62 down-regulated proteins and 67 up-regulated proteins,in the high-dose group of Xiaoai Jiedu Recipe compared with the model group,and these differential expressions were involved in lipid metabolism,et al.The results of biochemical tests showed that the intervention of Xiaoai Jiedu Recipe increased the serum iron and MDA levels,and decreased the GSH level in mice.Western blot results showed that the protein expression levels of Nrf2 and HMOX1 increased,and those of SLC7A11 and GPX4 decreased after the intervention of the high-dose group of Xiaoai Jiedu Recipe(P<0.01).CONCLUSION Xiaoai Jiedu Recipe pro-motes ferroptosis in hepatocellular carcinoma cells by inducing the Nrf2/HMOX1 signaling pathway,regulating oxidative stress and ele-vating the level of lipid peroxidation,which may be one of its mechanisms of action in inhibiting the growth of transplanted tumors.

Objective: To explore the anti-lung cancer mechanism of Maimendong Tang and Qianjin Weijingtang (Jin Fang) by detecting the expression profiles of long noncoding RNA (lncRNA) and mRNA in mice tumor tissues of orthotopic Lewis lung cancer model. Method: C57BL/6 mice were randomly divided into normal group, model group and Jin Fang group (20 g·kg^-1·d^-1). After successful establishment of Lewis lung cancer model in situ in mice, Jin Fang was given orally the next day after treatment. Using gene chip technology, differential lncRNA and mRNA closely related with Jin Fang' s anti-lung cancer effect were detected, and cluster analysis was performed. The key lncRNA and mRNA were screened out by t-test and fold change of differential expression. Bioinformatic methods were used to predict target genes regulated by differential lncRNA, and functional and pathway analysis was performed. The histopathological technique was used to detect the differences in the tumor tissue of each group under light microscope. Result: lncRNA and mRNA chip hybridization results showed that Jin Fang regulated differential expressions of 887 lncRNA, in which 442 were up-regulated and 445 were down-regulated (P<0.05). There were 610 differential mRNA expressions, in which 376 were up-regulated and 234 were down-regulated(P<0.05). GO analysis showed that down-regulated or up-regulated target genes were mainly involved in biological processes (BP), such as cell metabolism regulation and signal pathway regulation. Molecular function (MF) analysis showed such functions DNA binding, protein binding, receptor binding and transcription factor activity in down-regulated or up-regulated target genes. Target genes were involved in multiple biological processes, cellular components and molecular functions. Signal pathway predictions indicated that Jin Fang can regulate the Janus kinase/signal transducer and activator of tran-ions(JAK/STAT) and other signaling pathways closely related to the development of lung cancer. The results of histopathological examination confirmed that Jin Fang could significantly improve the pathological changes of lung tissues in the mice compared with the model group. Conclusion: Jin Fang may exert its anti-lung cancer effect by regulating the expressions of multiple lncRNAs and mRNAs, and down-regulating related signaling pathways.

Aim To determine the effect of cornel iri-doid glycoside ( CIG ) on human hepatocyte cell line (L-02) injured by D-galactosamine (GalN) and tumor necrosis factor-α( TNF-α) .Methods Firstly, CIG was extracted , separated and purified . Cell lesion model injured by D-GalN/TNF-αwas tested by MTT method.T-AOC, SOD, MDA and calcium ion concen-tration were taken as indicators to study the effects of CIG on L-02 cell injured by D-GalN/TNF-α.The ex-pression of p-PERK, p-eIF-2α, caspase-3 protein were detected by Western blot .Results 44 mg · L-1 D-GalN and 100 μg · L-1 TNF-αwere suitable for L-02 cell lesion model.CIG high, middle, low concentra-tion group could significantly increase the L-02 cell ac-tivity by 21％, 13％, 8％, respectively and SOD activity and T-AOC ability as well compared with model group.At the same time, they markedly reduced the MDA activity except the low concentration .Three con-centrations of CIG could reduce the expression of endo-plasmic reticulum stress related protein PERK , eIF-2αand apoptosis-associated protein caspase-3. Conclu-sions CIG could protect L-02 cells injured by D-GalN/TNF-α.Increasing the cellular antioxidant abili-ty, reducing the damage of endoplasmic reticulum stress and the expression of apoptosis-associated protein may be the possible mechanism .

Acute retinal necrosis syndrome (ARNS) is a group of eye syndrome.Acute uveitis, retinal artery occlusive vasculitis, fused necrotic retinitis and late stage of retinal detachment is the main clinical manifestation.A part of patients may be associated with increased intraocular pressure.The etiology and pathogenesis is still not clear completely and most people think that may be related to the virus infection, which mainly to reflected to be herpes simplex virus (HSV), varicella zoster virus (VZV), EB virus and giant cell virus (CMV) infection.Its diagnosis mainly depends on clinical manifestation, examination and etiological examination.Acute retinal necrosis syndrome is urgent and develops quickly, and it is lack of specific clinical symptoms in early times.By the way, it enjoys high misdiagnosis rate and poor prognosis.It is hard to cure, therefore, it is an important reason for the blindness.Once diagnosed, treatment should be adopted by carrying local and systemic antiviral, preventive laser photocoagulation in time.At the same time, it is essential that vitreous body resection combine with silicone oil tamponade treatment when necessary.The study shows that the effective measures of early treatment will be able to prevent disease progression and improve visual acuity.Therefore, early diagnosis and treatment of acute retinal necrosis syndrome is very important.In this paper, combination of the literature on the diagnosis and treatment of acute retinal necrosis syndrome were reviewed.

OBJECTIVETo discuss the mechanisms of electroacupuncture at "Neiguan" (PC 6) on p38 MAPK signaling pathway in rats with cardiac hypertrophy.

METHODSForty SD rats were randomly divided into four groups: a normal group, a model group, a model plus p38 MAPK inhibitor group, a model plus electroacupuncture group, ten rats in each group. The model rats were established by subcutaneous injection 3 mg/(kg x d) of Isoprenaline Hydrochloride; model plus p38 MAPK inhibitor group were injected 0.3 mg/(kg x d) of specific inhibitor SB 203580; model plus electroacupuncture group was treated by electroacupuncture at "Neiguan"(PC 6) with continuous-wave, 2 Hz and 1 mA for 20 minutes, once a day for 14 days. There was no treatment in other two groups. The contents of TNF-alpha and IL-1beta in heart tissue were detected by radioimmunoassay and the p38 MAPK, p-p38 MAPK by western blot.

RESULTSCompared with normal group, the contents of TNF-alpha, IL-1beta, p38 MAPK, p-p38 MAPK were significantly increased in model group (all P < 0.01). The contents of TNFalpha, IL-1beta, p38 MAPK, p-p38 MAPK were significantly decreased in model plus p38 MAPK inhibitor group and model plus electroacupuncture group, compared with model group, all P < 0.05; compared with normal group, P < 0.05, P < 0.01; but no significant difference between model plus p38 MAPK inhibitor group and model plus electroacupuncture group (P > 0.05).

CONCLUSIONElectroacupuncture at "Neiguan" (PC 6) can prevent the phosphorylation of p38 MAPK of myocardial hypertrophy, and the mechanism maybe adjust p38 MAPK signaling pathway by inhibiting the expression of TNF-alpha and IL-1beta.

Acupuncture Points ; Animals ; Cardiomegaly ; metabolism ; therapy ; Electroacupuncture ; Interleukin-1beta ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Tumor Necrosis Factor-alpha ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo study the expression of P57(kip2) and Maspin in the pathological scar and their possible role in the pathogenesis of abnormal scars.

METHODSImmunohistochemistry integrated image analysis and reverse transcription polymerase chain reaction (RT-RCR) were performed to detect the expression of P57(kip2) and Maspin in hypertrophic scar, keloid, mature scar and normal skin. Statistics was used to analyze the datas.

RESULTSThe expression of P57(kip2) protein was fixed to fibroblast intranuclear in abnormal scar, and the expression of P57(kip2) protein and P57(kip2) mRNA decreased (P < 0.05). The expression of Maspin protein was fixed to fibroblast cytoplasm and intranuclear in abnormal scar, and the expression of Maspin protein and Maspin mRNA decrease, compared with that in normal group (P < 0.05). There was positive correlation between P57(kip2) protein and Maspin protein expression (P < 0.01).

CONCLUSIONSThe decreased expression of P57(kip2) and Maspin in abnormal scar shows that they are cicatrix-related genes. There is a positive relationship between the two genes. It may be one of the mechanisms of pathogenesis of abnormal scar. It makes effect through fibroblasts.

Cicatrix ; metabolism ; pathology ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Cyclin-Dependent Kinase Inhibitor p57 ; metabolism ; Fibroblasts ; metabolism ; Humans ; Serpins ; metabolism

OBJECTIVETo discuss the reasons and the treatments for the femoral refracture.

METHODSFrom 2004 to 2008, 10 cases of femoral refracture after plate removal were selected and treated with open reduction and bone nail internal fixation, included 4 males and 6 females ranging from 19 to 48 years with an average age of 33 years old. According to Müller classification, there were 4 cases of type A, 3 of B, 3 of C. The skeletal tissues were taken for pathological analysis.

RESULTSTen patients were followed-up for 10 to 18 months (averaged 14 months). All wound healed at one stage,there were no complications. The fuction of lower limbs walking and weight loading werer recovered. Pathological section showed that part of the lamellar bone tissues were necrotic. The bone architectures were chaotic, micrangium blocked, Haversian canals were atrophy, new vessels and Haversian canals growed in necrotic bone. The tunnel in rigid bone and new Haversian system were formed. The structure was cutting cone. It was the process of new bone substitutting necrotic bone in bony remodeling.

CONCLUSIONThe internal fixation using ordinary plates overwhelmingly destroy blood circulation of bone and cause necrosis of bone. It can also effect bone remodeling and descent mechanical property. Refracture can readily happen after the plate removal. Treatment of open reduction and bone nail internal fixation can achieve excellent and good effect.

Adult ; Bone Plates ; Female ; Femoral Fractures ; pathology ; surgery ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Recurrence

Background and Objective: Chemotherapy regimen containing anthracyclines has been used as the standard treatment for acute myeloid leukemia (AML). This study was to compare the efficacy and toxicity of the chemotherapy regimen containing perarubicin (THP) with that containing mitoxantrone (MIT) for young patients with newly diagnosed AML. Methods: A total of 129 patients with newly diagnosed AML, aged 16 to 60 years olds, were assigned for induction chemotherapy containing one to two courses with standard-dose cytarabine (Ara-C) and an anthracycline antibiotic, THP or MIT. When complete remission was achieved after induction therapy, the patients received two courses of consolidation therapy identical to the induction regimen. From then, the patients were alternately given four courses of consolidation therapy consisting of Ara-C/I-HP or Ara-C/MIT every three weeks. Maintenance treatment continued for three years when patients were in continuous complete remission (CCR). Results: Twenty-six out of 42 patients (61.90%) receiving THP therapy, and 48 out of 73 patients (65.75%) treated by MIT achieved CR (P>0.05). Nine (34.61%) and 11 (22.92%) out of CR patients treated by THP and MIT, respectively, relapsed within one year (P= 0.28). Moreover, the incidences of toxicities, such as infection, nausea/ vomiting and cardiac events, were similar in these two groups (P>0.05) except for alopecie, which was 26.19% in the THP group compared to 42.47% in the MIT group (P<0.01). Conclusions: Regimen containing THP plus Ara-C can be used for young adults with newly diagnosed AML for remission induction, but it is not superior to the regimen with MIT. Consolidation chemotherapy with THP or MIT is feasible for young adults with AML after CR.

OBJECTIVETo compare the differences of the efficacy and different therapeutic drugs on the treatment of benign prostatic hyperplasia (BPH) in order to ensure the optimal indication for different BPH patients.

METHODSA randomized, parallel-controlled, multicenter clinical trial was conducted. From September 2002 to December 2003 906 BPH patients were enrolled into 7 therapeutic groups, including selective-adrenoceptor antagonist (terazosin, doxazosin tamsulosin and naftopidil), 5 alpha-reductase inhibitor (finasteride and epristeride) and natural product (cernilton). International Prostate Symptom Score (IPSS) and Quality of Life (QOL), uroflowmetry, total prostatic volume (TPV) and transitional zone volume and residual urine were used as efficacy criteria.

RESULTSAccording to the baseline, the IPSS and Qmax were significantly correlated to the prostatic volume and transitional zone volume (P < 0.01). At average follow-up of 6 months, significant improvements in IPSS, QOL, Qmax and residual urine volume were observed in each therapeutic group, and no difference in IPSS improvement was found among the groups. Prostatic volume and transitional zone volume were significant decreased in 5alpha-reductase inhibitor groups (P < 0.05). In patients with baseline TPV greater than 35.5 cm3, the improvement of Qmax was more significant than that in patients with TPV less than 35.5 cm3 in finasteride group (P < 0.01) (5.7 ml/s and 2.2 ml/s respectively), and more significant symptomatic improvements were also found in cernilton, doxazosin and naftopidil group. In each group, the improvement of symptom were more significant in patients with IPSS higher than 20 points (P < 0.01).

CONCLUSIONSEach drug observed in this study can improve the subjective and objective symptoms significantly for BPH patients, especially for patients with higher IPSS baseline. When using 5alpha-reductase inhibitor, prostatic volume can be decreased significantly and more obviously subjective and objective improvement can be found in the patients with TPV greater than 35.5 cm3.

5-alpha Reductase Inhibitors ; Adrenergic alpha-Antagonists ; therapeutic use ; Aged ; Aged, 80 and over ; Androstadienes ; therapeutic use ; Double-Blind Method ; Doxazosin ; therapeutic use ; Enzyme Inhibitors ; therapeutic use ; Finasteride ; therapeutic use ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Naphthalenes ; therapeutic use ; Piperazines ; therapeutic use ; Plant Extracts ; therapeutic use ; Prazosin ; analogs & derivatives ; therapeutic use ; Prostate ; drug effects ; pathology ; physiopathology ; Prostatic Hyperplasia ; drug therapy ; Quality of Life ; Secale ; Sulfonamides ; therapeutic use ; Treatment Outcome

Adenovirus vectors are one of the most promising gene transfer systems. They are of great value for gene therapy because these vectors achieve temporal high-level transgene expression and high gene transfer efficiency. To meet increasing needs of adenovirus vectors for gene therapy programs, parallel development of efficient, scalable and reproducible production processes is required. Perfusion cultivation of 293 cells is one of the most commonly used methods to produce adenovirus vectors and it is suitable for industrialized production specially. Experimental studies had been carried out to produce recombinant adenovirus containing the green fluorescent protein gene (Ad-GFP) by perfusion cultivation of HEK-293 N3S cells in a 5L stirring bioreactors. Perfusion rate was 1-2 volume/day. To infect the 293 N3S cells with Ad-GFP at the density of (2-4) x 10(6) cells/ ml. The time of collecting cells was 48 hours post infection. After three rounds of freeze/thaw and centrifugation, the crude viral lysates were stored at--80 degrees C until use. Then to get the Ad-GFP products by 2 x CsCl-gradient purification. The purity of the products was determined by the A260/A280 ratio and a high performance liquid chromatography (HPLC) assay. The infective titer was determined by a TCID50 assay. The culture term was 10-12 days. The infectious titer, the number of virus particle and the ratio of infectious titer to virus particle for the product were 1.0 x 10(11) IU/mL, 1.68 x 10(12) VP/mL and 6.0% IU/VP respectively. The A260/A280 ratio was 1.33, and the purity determined by HPLC was 99.2%. The cell specific productivity was around 1000 IU/cell. By perfusion cultivation of 293 N3S cells in a 5L stirring bioreactors, we established the production process for Ad-GFP, which paves a way to produce other recombinant adenovirus for gene therapy.
Adenoviridae ; genetics ; growth & development ; isolation & purification ; Bioreactors ; microbiology ; Cell Line ; Gene Transfer Techniques ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Kidney ; cytology ; virology ; Recombinant Proteins ; biosynthesis ; genetics ; Recombination, Genetic ; Virus Cultivation ; instrumentation ; methods