A Highly Sensitive Spectrophotometric Assay of Bleomycins Based on the Fading Reaction of Some Halofluorescein Dyes

In weak acidic medium, anticancer antibiotics bleomycin A5 （BLMA5） and bleomycin A2 （BLMA2） can react with halofluorescein dyes such as erythrosin （Ery）, eosin Y （EY）, eosin B （EB） and rose bengal （RB） by virtue of electrostatic attraction and hydrophobic force to form the ion-association complexes, which can result in the fading reactions of four halofluorescein dyes. The maximum fading wavelengths of these four dyes were located at 527 nm for Ery, 515 nm for EY, 517 nm for EB and 546 nm for RB, respectively. The decrements of absorbance （AA） were directly proportional to the concentrations of bleomycin in a certain range. A new method for the determination of bleomycins anticancer drugs based on fading reactions of halofluorescein dyes has been developed. The method was not only highly sensitive but also simple and rapid. The molar absorptivities （ε） ranged from 1.5 × 10^5 to 7.5 × 10^5 L·mol^-1·cm^-1. It was applied to determination of the bleomycins in human serum, urine and rabbit serum samples. In this work, the spectral properties and the optimum reaction conditions were investigated. The structure of ion-association complexes and the reaction mechanism were discussed.

Trigonelline mitigates bleomycin-induced idiopathic pulmonary fibrosis in mice

Objective:To evaluate the effect of trigonelline on bleomycin-induced idiopathic pulmonary fibrosis(IPF)and to explore its underlying mechanisms using network pharmacology.Methods:IPF was induced in C57BL/6 mice by a single intratracheal instillation of bleomycin(5 mg/kg).Trigonelline was administered at doses of 25,50,and 100 mg/kg/day orally from the 2nd day post-bleomycin induction up to the 14th day.In IPF-induced mice,lung coefficient,immune cell infiltration in bronchoalveolar lavage fluid,and oxidative stress were measured.Histological alterations in lung tissues were also assessed.Moreover,network pharmacology approach was conducted to reveal molecular interactions of bleomycin and trigonelline with targets of IPF.Results:Trigonelline treatment reduced bleomycin-induced oxidative stress and immune cell infiltration,and mitigated physiological changes in the lung tissues of mice.Moreover,trigonelline alleviated bleomycin-induced histological alterations in lung tissues.Network pharmacology analysis showed that bleomycin and trigonelline interacted with IPF targets,such as NFKB1,HDAC2,HIF1A,and TLR4.Conclusions:The interaction of trigonelline with key IPF targets and its ameliorative effects on lung damage and oxidative stress highlight its potential in treating IPF.It may be considered an antifibrotic agent for further clinical development.

Investigating the mechanism of action of Bu-Yang-Huan-Wu decoction in treating bleomycin-Induced pulmonary fibrosis through the epithelial-mesenchymal transition pathway

Background:To explore the effects and mechanisms of Bu-Yang-Huan-Wu Decoction on pulmonary fibrosis in mice.Methods:Forty-five C57BL/6J mice were randomly divided into three groups:Control,Model,and Bu-Yang-Huan-Wu Decoction.Pulmonary fibrosis was elicited in mice through a solitary intratracheal administration of 2.5 mg/kg bleomycin.For the control group,mice were given a solitary intratracheal administration of a comparable volume of PBS.Treatment began on the first day after the successful model establishment and lasted for 21 days.The survival rate and body weight of the mice were recorded daily,and on the 22nd day,bronchoalveolar lavage fluid was collected to determine total cells and total protein.The wet/dry weight ratio of lung tissue and hydroxyproline were measured.Lung tissue pathology was observed using hematoxylin and eosin staining and Masson staining.The mRNA expression of epithelial-mesenchymal transition-related proteins(E-cadherin and vimentin)was detected by RT-qPCR,and their protein expression was analyzed by western blot.Results:Compared to the model group,the Bu-Yang-Huan-Wu Decoction treatment notably enhanced both the survival rate and body weight in pulmonary fibrosis mice,significantly reduced lung tissue wet/dry weight ratio,total cells,and protein in bronchoalveolar lavage fluid,and hydroxyproline content.The pathological morphology of lung tissue was significantly improved,with increased expression of the epithelial cell marker E-cadherin mRNA and protein,and decreased expression of the mesenchymal cell marker vimentin mRNA and protein.Conclusion:Bu-Yang-Huan-Wu Decoction can improve the degree of bleomycin-induced pulmonary fibrosis in mice by inhibiting epithelial-mesenchymal transition.

博来霉素的抗肿瘤活性及诱导肺纤维化毒性的研究进展

博来霉素(bleomycin,BLM)是日本科学家Umezawa在1966年从轮枝链霉菌(Streptomyces verticillus)中首次分离得到一种糖肽类天然抗生素。研究发现博来霉素具有抗癌活性,在临床上作为一类化疗药物,广泛用于治疗头颈部鳞癌、鼻咽癌、食管癌、恶性淋巴瘤、乳腺癌、绒毛膜癌等,与顺铂、依托泊苷联合使用,对睾丸癌的有效率约为90%[1]。尽管博来霉素具有抗瘤作用强、抗瘤谱广、给药途径多的优点,通常不会造成患者的白细胞减少和抑制免疫功能。但是文献报道博来霉素诱发约46%患者发生肺炎样症状及肺纤维化症状,其中3%的病人会发生死亡[2],从而大大限制了博来霉素的使用。博来霉素的化学结构已经明确,其金属结合区、连接区、联噻唑端基区在抗肿瘤方面发挥不同作用,结构中二糖部分的生物活性是目前的研究热点。

LOW DOSE PIRFENIDONE SUPPRESSES TRANSFORMING GROWTH FACTOR BETA-1 AND TISSUE INHIBITOR OF METALLOPROTEINASE-1, AND PROTECTS RATS FROM LUNG FIBROSIS INDUCED BY BLEOMYCIN

Objective To investigate the optimal dosage of pirfenidone for the treatment of pulmonary fibrosis induced by bleomycin in Wistar rats, and the alteration of expressions of transforming growth factor beta-1 （ TGF-β1 ）, tissue inhibitor of metalloproteinase-1 （ TIMP-1 ）, and matrix metalloproteinase-13 （ MMP-13 ） in lung tissue. Methods Male Wistar rats were endotracheally instilled with bleomycin or normal saline. Pirfenidone （25-800 mg · kg^-l · d^-1 ）, dexamethasone （3 mg/kg）, or 1% carboxymethylcellulose sodium were given daily by feed 2 days before instillation of bleomycin. Groups T7 and T14 were fed pirfenidone 50 mg · kg^-1 · d^-1 at 7 days or 14 daYs after bleomycin instillation. Lungs were harvested at 28 days after bleomycin instillation. Patholological changes in luffg tissues were evaluated with HE staining. Lung collagen was stained by sirius red and measured by content of hydroxypro- line. Expression of proteins of TGF-β1 TIMP-1, and MMP-13 were detected by Western blotting. Results At doses of 25, 50, and 100 mg· kg^- 1 · d ^- 1, pirfenidone had significant anti-fibrotic effects for bleomy- cin-induced rat pulmonary fibrosis, and these effects were most significantly attenuated at the dosage of 50 mg · kg^-1 ·d^ -1（ HE： P 〈 0. 01, P 〈 0.01, and P = 0.064; sirius red： P 〈0.05, P 〈 0.01, and P 〈 0.05 ; hydroxyproline： P = 0.595, P 〈 0.01, and P = 0.976）. Pirfenidone at a dosage of 50 mg · kg^- l · d^-1 inhibited protein expression of TGF-131 and TIMP-1 in lung tissue in the early phase （0.79 and 0.75 times of control group）, but had no effect on ex- nr^eelnn nf MMP-13. Conclusion Low dose pirfenidone, especially at dosage of 50 mg · kg^-1 · d^-1, has significant anti-fibrotic effects on bleomycin-induced rat pulmonary fibrosis. Pirfenidone partially inhibits the enhancement of the expression of TGF-131 and TIMP-β1 in lung tissue.

A novel pulmonary fibrosis murine model with immune-related liver injury

Idiopathic pulmonary fibrosis(IPF),characterized by aggravated alveolar destruc-tion and fibrotic matrix deposition,tendentiously experiences the stage called acute exacerbation IPF(AE-IPF)and progresses to multiple organ damage,especially liver injury.Recent studies have found a variety of immune microenvironment disorders associated with elevated IPF risk and secondary organ injury,whereas current animal models induced with bleomycin(BLM)could not completely reflect the pathologi-cal manifestations of AE-IPF patients in clinic,and the exact underlying mechanisms are not yet fully explored.In the current study,we established an AE-IPF model by tracheal administration of a single dose of BLM and then repeated administrations of lipopolysaccharide in mice.This mouse model successfully recapitulated the clinical features of AE-IPF,including excessive intrapulmonary inflammation and fibrosis and extrapulmonary manifestations,as indicated by significant upregulation of Il6,Tnfa,Il1b,Tgfb,fibronectin,and Col1a1 in both lungs and liver and elevated serum aspartate transaminase and alanine transaminase levels.These effects might be attributed to the regulation of Th17 cells.By sharing this novel murine model,we expect to pro-vide an appropriate experimental platform to investigate the pathogenesis of AE-IPF coupled with liver injury and contribute to the discovery and development of targeted interventions.

Activity of boanmycin against colorectal cancer

INTRODUCTIONBoanmycin (Bleomycin A6, BAM ), a newantitumor antibiotic, was isolated from manycomponents of bleomycin (BLM) produced bystreptomyces pingyangensis which were obtainedfrom a soil sample collected in Pingyang County,Zhejiang Province, China. Boanmycin has a similarchemical structure to that of BLM, but the terminalamine moiety is different[ 1].

Modulatory effect of D-pinitol on bleomycin-induced pulmonary fibrosis in rats

Objective:To assess the effect of D-pinitol on pulmonary fibrosis induced by bleomycin.Methods:Sprague-Dawley rats received intratracheal bleomycin(6 IU/kg)to induce pulmonary fibrosis,followed by administration of either D-pinitol(5,10,or 20 mg/kg)or vehicle or methylprednisolone(10 mg/kg)over 28 days after bleomycin administration.Lung function,biochemical parameters,serum biochemistry,mRNA expressions,and histological features were observed.Results:D-pinitol at 10 and 20 mg/kg significantly(P<0.05)attenuated bleomycin-induced bronchoalveolar lavage fluid,decreased myeloperoxidase,nitric oxide,malondialdehyde levels,and increased glutathione and superoxide dismutase level.D-pinitol also improved lung function(enhanced pause,frequency of breathing,expired volume,and tidal volume).Besides,D-pinitol significantly(P<0.05)upregulated Nrf2 and downregulated mRNA expressions of TGF-β,collagen-1,and Smad-3.Furthermore,considerably less inflammation(peribronchial,perivascular,and total),Ashcroft,and interstitial fibrosis scores were observed in the D-pinitol group.Conclusions:D-pinitol exerts its effect against bleomycin-induced pulmonary fibrosis via antioxidative and anti-fibrotic pathways.

Cyclophosphamide abrogates the expansion of CD4^(+)Foxp3^(+) regulatory T cells and enhances the efficacy of bleomycin in the treatment of mouse B16-F10 melanomas

Objective:Promotion of the proliferative expansion of CD4^(+)Foxp3^(+)regulatory T cells(Tregs)is one of the side effects that limits the use of bleomycin(BLM)in the treatment of tumors.In this study,we examined the hypothesis that cyclophosphamide(CY),a chemotherapeutic agent with the capacity to eliminate tumor infiltrating Tregs,abrogated BLM-induced expansion of Tregs and consequently resulted in a better anti-tumor effect.Methods:The in vitro effects of BLM,with or without mafosfamide(MAF,the active metabolite of CY),on both TGF-β-induced differentiation of Tregs(iTregs),and TNF-induced expansion of naturally occurring Tregs(nTregs)were assessed.The in vivo effect of low doses of BLM and CY on tumor-infiltrating Tregs,as well as on the growth of mouse B16-F10 melanomas,was also studied.Results:In vitro treatment with BLM promoted the differentiation of iTregs,as well as TNF-induced expansion of nTregs.These effects of BLM were completely abrogated by MAF.Furthermore,in the mouse B16-F10 melanoma model,treatment with low doses of BLM increased the number of tumor-infiltrating Tregs,and this effect of BLM was also abrogated by CY.Importantly,combination therapy with low doses of BLM and CY showed synergistic anti-tumor effects.Conclusions:CY abrogated the effect of BLM on the expansion of Tregs.The combination of these 2 chemotherapeutic agents may represent a safer and more effective therapy in the treatment of cancer patients,and thus merits future clinical evaluation.

Early- and intermediate-term outcome of transarterial embolization for symptomatic hepatic focal nodular hyperplasia

Purpose: To evaluate the early-and intermediate-term outcome in patients with symptomatic hepatic focal nodular hyperplasia(FNH) treated with transarterial embolization using bleomycin-iodinated oil and polyvinyl alcohol(PVA) particles. Materials and methods: In this two-center retrospective study between January 2005 and December 2013, 27 consecutive patients with symptomatic hepatic FNH underwent superselective catheterization by microcatheter techniques and embolization using bleomycin-iodinated oil combined with PVA. Early-term(3–41 months) follow-up of TAE was performed in terms of symptom control, changes in lesion size, and complications. Intermediate-term(45–112 months) follow-up was carried out to assess symptom control and reinterventions for recurrence. Results: Embolization was performed in 27 patients with 31 lesions. Technical success was achieved in all cases. The follow-up period ranged from three to 112 months. At early-term follow-up, mean lesion diameters were decreased significantly from 6.4±2.7 cm pre-intervention to 3.6±1.8 cm at 3-9 months after embolization(P<0.001). A total of 7 lesions had complete resolution during the whole follow-up period. At intermediate-term follow-up, local recurrence was found in 1 treated lesion at 54-months. Contrast-enhanced scans showed complete lack of residual arterial blood supply in the majority of lesions during the follow-up period. There was no major complication associated with the procedure. Conclusion: Transarterial embolization using bleomycin-iodinated oil and PVA is a feasible, safe and effective alternative in both early-and intermediate-terms for the treatment of symptomatic hepatic FNH.

Treatment of Interstitial Peumopathy by Fei Tong Oral Liquid in the Malignant Tumor Patients after Radio-and/or Chemotherapy

Fei Tong Kou Fu Ye (肺通口服液 Fei Tong Oral Liquid) was used to treat 30 cases of interstitial pneumopathy after radio- and/or chemotherapy.In comparison with the control group (15 cases) treated with hormones,the therapeutic effects in improving dyspnea,cough,respiratory rate,cyanosis,findings in X-films and CT examination,partial pressure of oxygen in artery,FVC and VC were found significantly better (P<0.05).The total effective rate obtained was 83.33%.

SRT1720, a SIRT1 Activator, Aggravates Bleomycin-Induced Lung Injury in Mice

Diagnosis and management of interstitial lung diseases (ILDs), caused by lung epithelial injury followed by apoptosis, are often challenging. It has been controversial whether the SIRT1 protein, a principal modulator of longevity due to caloric restriction, ameliorates or aggravates ILD in animal models. Here we examined the effect of SRT1720, a syn- thetic activator of SIRT1, on bleomycin-induced lung injury in a mouse model and apoptosis in cultured epithelial cells. Oral intubation of SRT1720 over a period of 15 days caused body weight loss and a high mortality rate among bleomy- cin-treated mice. Histological examinations showed that the SRT1720 load increased fibrosis in the bleomycin-treated lung. An analysis of bronchoalveolar lavage fluid revealed remarkably increased numbers of inflammatory cells in the SRT1720-treated group. Moreover, the apoptosis of A549 lung cancer cells, caused by X-ray irradiation and an anti-Fas activating antibody, was promoted by SRT1720. These results indicate that SRT1720 not only aggravates bleomy- cin-induced ILD, but stimulates the apoptosis of physically and biologically stimulated A549 cells. While SIRT1 acti- vators are considered promising for the treatment of conditions such as diabetes mellitus, fatty liver, and chronic ob- structive pulmonary diseases, an excess of food containing SIRT1 activators may be harmful depending on the disease state, especially in the case of acute inflammation.

Expression of PEPT2 mRNA in the lung of rat with bleomycin-induced pulmonary fibrosis

Objective：Pulmonary fibrosis is a common pathological phenomena in lung cancer patients after chemotherapy or radiotherapy, which is a key factor hindering to transport ion of high concentrated drug to the lung tissue, peptide trans-porter has become targets of the rational design of peptides and peptide drug. The purpose of the study is to investigate the expression of PEPT2 mRNA in the lung of rats with bleomycin （BLM）-induced pulmonary fibrosis. Methods：Fifty healthy adult Spragne-Dawley rats were randomized into five groups, the rats in BLM 7d, 14d and 28d groups were treated with a single instil ation of 5 mg/kg of BLM, to induced pulmonary fibrosis models. On days 7, 14 and 28, the animals were kil ed by exsan-guination respectively. Normal saline （NS） group were treated by NS, on days 14, the animals were kil ed by exsanguinations. Control group were untreated. The lung samples were processed for light microscopy and determined the hydroxyproline （HYP） concentration. The expression of PEPT2 mRNA were measured by RT-PCR. PEPT2 cDNA fragments were tested by dideoxy chain termination. Results：Compared with control and NS group, HYP levels increased on day 7 of BLM group, but there was no statistical significant dif erence （P〉0.05）. HYP levels markedly increased on days 14 and 28 of BLM group, there was statistical significant dif erence （P〈0.01）. The morphological study showed that col agenous fiber proliferated on days 14 and 28 of BLM group, especial y on day 28, formed pulmonary fibrosis. There were no significant changes of pulmo-nary PEPT2 mRNA expression at dif erent groups （P〉0.05）. Conclusion：The pulmonary fibrosis models of SD rats can be induced by a single instil ation of 5 mg/kg of bleomycin on 28d. There were no significant changes of PEPT2 mRNA expression in the lung of rats with bleomycin-induced pulmonary fibrosis.

Effects and possible anti-tumor immunity of electrochemotherapy with bleomycin on human colon cancer xenografts in nude mice

AIM: To evaluate the anti-tumor effects and possible involvement of anti-tumor immunity of electrochemotherapy (ECT) employing electroporation and bleomycin in human colon cancer xenografts in nude mice, and to establish the experimental basis for clinical application of ECT. METHODS: Forty nude mice, inoculated subcutaneously human colon cancer cell line LoVo for 3 wk, were allocated randomly into four groups: B+E+ (ECT), B+E- (administration of bleomycin alone), B-E+ (administration of electric pulses alone), and B-E- (no treatment). Tumor volumes were measured daily. The animals were killed on the 7* d, the weights of xenografts were measured, and histologies of tumors were evaluated. Cytotoxicity of spleen natural killer (NK) and lymphokine-activated killer (LAK) cells was then assessed by lactic dehydrogenase release assay. RESULTS: The mean tumor volume of group B+E+ was statistically different from the other three groups after the treatment (F= 36.80, P<0.01). There was one case of complete response, seven cases of partial response (PR) in group B+E+, one case of PR in group B+E- and group B-E+ respectively, and no response was observed in group B-E-. The difference of response between group B+E+ and the other three groups was statistically significant (X2 = 25.67,P<0.01). Histologically, extensive necrosis of tumor cells with considerable vascular damage and inflammatory cells infiltration were observed in group B+E+. There was no statistical difference between the cytotoxicity of NK and LAK cells in the four treatment groups. CONCLUSION: ECT significantly enhances the chemosensitivity and effects of chemotherapy in human colon cancer xenografts in nude mice, and could be a kind of novel treatment modality for human colon cancer. The generation of T-cell-dependent, tumor-specific immunity might be involved in the process of ECT.

CXC CHEMOKINE RECEPTOR 3 MODULATES BLEOMYCIN-INDUCED PULMONARY INJURY VIA INVOLVING INFLAMMATORY PROCESS

Objeelive To investigate the role of CXC chemokine receptor 3 （ CXCR3 ） in bleomycin-induced lung injury by using CXCR3 gene deficient mice. Methods Sex-, age-, and weight-matched C57BL/6 CXCR3 gene knockout mice and C57BL/6 wide type mice were challenged by injection of bleomycin via trachea. Lung tissue was stained with HE method. Airway resistance was measured. Bronchoalveolar lavage （BAL） was performed using phosphate buffered saline twice, cell number and differentials were counted by Diff-Quick staining. Interleukin （IL）-4, IL-5, IL-12p40, and interfon-~ in BAL fluid and lung homogenate were measured by enzyme-linked immunosorbent assay. Unpaired t test was explored to compare the difference between two groups. Results On day 7 after bleomycin injection via trachea, CXCR3 knockout mice were protected from bleomycininduced lung injury as evidenced by fewer accumulation of inflammatory cells in the airway and lung interstitium compared with their wild type littermates （ P 〈 0. 05 ）. Airway resistance was also lower in CXCR3 knockout mice compared with wild type mice （P 〈 0. 01 ）. Significantly lower level of inflammatory cytokines release, including the altered production of IL-4 and IL-5 both in BAL fluid and lung tissue was seen in CXCR3 knockout mice than in wild type mice （both P 〈0. 05）. Conclusion CXCR3 signaling promotes inflammatory cells recruiting and initiates inflammatory cytokines cascade following endotracheal bleomycin administration, indicating that CXCR3 might be a therapeutic target for pulmonary injury.

The Electrochemical Behavior of Bleomycin at A Co/GC Ion Implantation Modified Electrode

In 0.10 mol/L HOAc-NaOAc buffer solution (pH 4.59). a sensitive reduction peak of bleomycin is obtained by linear sweep voltammetry at Co/GC ion implantation modified electrode. Its electrochemical behavior has been studied. The experiments of AES and XPS show that Co is surely implanted into the surface of GCE and improved the electrocatalytic activity.

VEGFmRNA and eNOSmRNA expression in immature rabbits with bleomycin-induced pulmonary hypertension

Objective: To investigate the evolution of pulmonary hypertension, the pathological changes of pulmonary arteries, and the expression of VEGFmRNA and eNOSmRNA of pulmonary arterial endothelial cells in immature rabbits treated with intratracheal bleomycin (BLM). Methods: Immature rabbits were divided into control and BLM group. Two and four weeks after intratracheal normal saline or BLM injection, the systolic, diastolic and mean pulmonary artery pressure (PASP, PADP, MPAP) were measured by micro-catheter; the pathological changes and the expression of VEGFmRNA and eNOSmRNA of endothelial cells in pulmonary arteries were evaluated by HE and in situ hybridization.Results: Two and four weeks after intratracheal injection of BLM, the PASP, PADP and MPAP increased 53%, 49%, 52%in 2 weeks, and 43%, 89%, 56% in 4 weeks; the wall thickness increased and the cavity in middle and small pulmonary arteries became narrow; the Thickness Index (TI) and Area Index (AI) increased 25%, 14% in 2 weeks, and 22%, 24% in 4 weeks; the level of VEGFmRNA and eNOSmRNA expression decreased 46%, 43% in 2 weeks, and 43%, 51% in 4 weeks.There was no significant difference between 2 weeks and 4 weeks BLM groups. Conclusion: The pulmonary artery pressure was elevated, the thickness of wall increased and the cavity became narrow in middle and small pulmonary arteries, and the level of VEGFmRNA and eNOSmRNA expression in pulmonary arterial endothelial cells decreased in immature rabbits after 2 weeks and 4 weeks of intratracheal 4 U/kg BLM injection.

Studies on the Electrochemical Behavior of Bleomycin at a Co/GC Ion Implantation Modified Electrode and Its Application

In a 0. 10 mol/L HAc-NaAc buffer solution (pH = 4. 59), a sensitive reduction peak of bleomycin was observed by linear sweep voltammetry at a Co/GC ion implantation modified electrode. The peak potential was-0. 73 V(iw. SCE). The peak current was proportional to the concentration of bleomycin over the range of 5.0 × 10-8-1.0× 10-6 and 1.0× 10-6-1. 0 × 10-5 mol/L with a detection limit of 2.0 × 10-8 mol/L. The electrochemical behavior of the reduction peak of bleomycin at the Co/GC modified electrode was studied by linear sweep and cyclic voltammetry and applied to the determination of bleomycin in urine. This method is simple, rapid and reliable. The reduction process is quasi-reversible. The experiments of AES and XPS showed that Co was surely implanted into the surface of GCE and the depth distribution of Co was in good agreement with Gooses normal distribution; the implanted Co at GCE improved the electrocatalytic activity.

ANTITUMOR ACTIVITY OF IMMUNOCONJUGATES COMPOSED OF BOANMYCIN AND MONOCLONAL ANTIBODY

Boanmycin (bleomycin A6 . BM) . an antitumor antibiotic, was conjugated to monoclonal antibodies including R19, H 111 and CCT2. The immunoconjugates exhibited selective cytotoxicity to related target cells including cecum cancer Hce-8693 cells, liver cancer BEL-7402 cells and leukemia CEM cells. They were highly effective against related human tumor xenografts in nude mice, and the inhibition rates by the conjugates were much higher than those by free BM. The inhibition rate by R19-BM conjugate against human cecum cancer xenografts reached 90%. BY immunoelectron microscopy, CCT2-BM conjugate showed specific binding and internalization in leukemia CEM cells. The results indicate that boanmycin-monoclonal antibody immunoconjugates are highly active both in vitro and in vivo.

Assessment of traditional Chinese medicine pattern in a bleomycininduced pulmonary fibrosis mouse model: A pilot study

Objective:To initially explore traditional Chinese medicine patterns in a bleomycin-induced pulmonary fibrosis mouse model.Methods:Thirty-six C57BL/6 mice were divided by the random number table method(with 12 rats per group)into three groups:a blank group,a model group,and a number 2 Feibi recipe(FBR-2)group.The pulmonary fibrosis mouse model was established by intratracheal instillation of bleomycin.The FBR-2 group was treated with FBR-2 for 4 weeks.Symptoms in the mice such as mental behavior,food/water intake,body weight,body temperature,respiratory rate,and tongue image were observed.The samples were collected on the 14th day and 28th day after modeling,and lung tissues were visually assessed and microscopically evaluated by staining with hematoxylin-eosin and Masson.The expression levels of hydroxyproline,interleukin(IL)-33,IL-37,tissue plasminogen activator,and plasminogen activator inhibitor-1 were determined by enzyme-linked immunosorbent assay.Results:Mice in the model group were poor in spirit,less active,slow in response,showed reduced food/water intake,body temperature,and body weight,increased respiratory rate,and their tongue color had changed from light red to dark red.However,treatment with FBR-2 significantly improved these symptoms.Extensive inflammatory cell infiltration and collagen fiber deposition were observed in the lung tissues of the model group.Compared with the blank group,the levels of hydroxyproline,IL-33,and plasminogen activator inhibitor-1 in the model group significantly increased(all P<.05),whereas that of tissue plasminogen activator significantly decreased on the 14th day and 28th day(P=.036 and P=.005,respectively).Moreover,FBR-2 improved lung inflammation and fibrinolysis imbalance and reduced collagen fiber deposition.Conclusion:To some extent,our bleomycin-induced pulmonary fibrosis mouse model exhibited traditional Chinese medicine patterns of qi deficiency,blood stasis,and heat retention.