Cell line模式在生产中的选择和应用

现代消费电子产品市场竞争激烈,客户需求呈现多样化发展,生产商需要根据不同机种的特点在皮带线模式和Cell line模式之间进行人力,设备,产能的评估并做出合理的选择和应用,从而可以在利润率不断下降的市场上稳定收益。     

Phase II Evaluation of VDC‐1101 in Canine Cutaneous T‐Cell Lymphoma

Background

Canine cutaneous T‐cell lymphoma (CTCL) is an uncommon disease for which efficacious therapies are lacking. The novel anticancer nucleotide prodrug VDC‐1101 (formerly known as GS‐9219) has shown efficacy in dogs with multicentric lymphoma. One of the observed adverse effects with this drug was a skin change characterized by hair loss, erythema, and pruritus, implying delivery of VDC‐1101 to the skin.

Hypothesis/Objectives

The primary study objective was to identify the objective response rate (ORR) to VDC‐1101 in canine CTCL; secondary objectives included characterization of progression‐free survival (PFS) and adverse events (AEs).

Animals

Twelve dogs with chemotherapy‐naïve or relapsed, histologically and immunohistochemically confirmed CTCL.

Methods

Dogs received VDC‐1101 as a 30‐minute IV infusion once every 21 days. Prednisone (1 mg/kg PO q48h) was administered concurrently.

Results

In 11 evaluable patients, responses included 1 complete response (CR), 4 partial responses (PR), 2 stable disease (SD), and 4 progressive disease for an ORR of 45% and biologic response rate (CR/PR/SD) of 64%. The median PFS was 37.5 days (26 to >399 days), which includes 1 durable and ongoing CR (>1 year). Gastrointestinal and hematologic AEs were mild; no dogs developed grade 3 or 4 AEs. Three dogs developed dermatopathies and 1 of these dogs was removed from the study as a result of this AE.

Conclusions and Clinical Importance

VDC‐1101 has activity against canine CTCL and could provide another treatment option in a disease process with a poor prognosis.     

Analysis of Seroreactivity against Cell Culture–Derived Bartonella spp. Antigens in Dogs

Background

Little is known about the specificity of Bartonella spp. immunofluorescent antibody (IFA) assays in dogs. Bacteremia in sick dogs most often has been associated with Bartonella henselae (Bh), Bartonella vinsonii subspecies berkhoffii (Bvb), and Bartonella koehlerae (Bk). Clarification of the diagnostic utility of IFA serology when testing against these organisms is needed.

Objective

To evaluate the specificity of Bartonella IFA assays utilizing 6 cell culture–grown antigen preparations.

Animals

Archived sera from SPF dogs (n = 29) and from dogs experimentally infected with Bvb (n = 10) and Bh (n = 3).

Methods

Antibodies (Abs) to Bvb genotypes I, II, and III, Bh serotype I, strains H‐1 and SA2, and to Bk were determined by IFA testing.

Results

Serum from naïve SPF dogs shown to be negative for Bartonella bacteremia did not react with any of the 6 Bartonella antigens by IFA testing. Dogs experimentally infected with Bvb genotype I developed Abs against homologous antigens, with no cross‐reactivity to heterologous Bvb genotypes, Bh H‐1, SA2 strains, or to Bk. Dogs experimentally infected with Bh serotype I developed Abs against Bh H‐1, but not to Bh SA2 strain with no cross‐reactive Abs to Bvb genotypes I–III or to Bk.

Conclusions and Clinical Importance

Bartonella spp. Ab responses during acute experimental infections are species and type specific.     

家蚕转基因高产丝研究论文被《Cell Research》列为研究亮点并专题评论

中科院上海生命科学研究院李胜研究员和西南大学夏庆友教授及其研究团队合作完成的家蚕转基因研究成果"Ras1CAoverexpressionin the posterior silk glandi mproves silk yield"     

Study on Expression and Immune Effect of F Protein of Newcastle Disease Virus Type Ⅶ in Insect Cell

This experiment was conducted to study the protein expression of F gene of Newcastle disease virus (NDV) type Ⅶ by baculovirus expression system.The F gene was amplified by RT-PCR and cloned into pFastBac HT A plasmid, and then the recombinant plasmid was transformed into DH10Bac competent cells to get the positive recombinant bacmid.After 72 h transfection of Sf9 cell with recombinant bacmid, the expression of interest protein was detected by indirect immunofluorescence assay (IFA), SDS-PAGE analysis and Western blotting. Serum antibody titers of immunized SPF chickens were determined by ELISA and the protective properties were determined by protection test. The results showed that F gene was expressed in Sf9 cells infected with the recombinant baculovirus. The expressed protein could induce high titer specific antibody against NDV. The protective rate of recombinant F protein group was 90%, which was significantly higher than that of the negative control group. These results laid a foundation for study on NDV subunit vaccine.     

Concordance of c‐kit Mutational Status in Matched Primary and Metastatic Cutaneous Canine Mast Cell Tumors at Baseline

Background

Mutation analysis of proto‐oncogene c‐kit (c‐kit) is advisable before starting treatment with tyrosine kinase inhibitors in dogs with mast cell tumor (MCT), including those with metastatic disease. Testing is usually performed on primary tumors, assuming that c‐kit mutation status does not change in metastasis.

Hypothesis/Objectives

To give an insight into the mutational processes and to make a recommendation on the use of c‐kit mutational analysis in the clinical setting.

Animals

Twenty‐one client‐owned dogs with metastatic MCT.

Methods

Dogs undergoing resection or biopsy for both primary and matched metastatic MCT were prospectively enrolled. Total RNA or DNA was extracted from primary MCT and corresponding metastases. Exons 8, 9, and 11 were amplified by PCR and sequenced. Genetic features between primary MCT and metastases were compared. Their correlation with clinicopathologic features was investigated.

Results

Concordance (mutated or wild‐type) of mutational status, evaluable in 21 primary and matched metastatic (20 nodal and 1 splenic) MCTs, was 100%. Three new c‐kit mutations were identified. No significant correlation was detected between c‐kit mutation and clinicopathologic features.

Conclusions and Clinical Importance

Proto‐oncogene c‐kit mutational status is conserved between any primary and its matched secondary tumor, suggesting that both can be used for c‐kit mutational testing. Targeted therapies might be also used to treat metastatic disease.     

Cats with Inflammatory Bowel Disease and Intestinal Small Cell Lymphoma Have Low Serum Concentrations of 25‐Hydroxyvitamin D

Background

Inflammatory bowel disease (IBD) and intestinal small cell lymphoma (ISCL) are common diseases in cats. The prevalence of alterations in the serum concentrations of fat soluble vitamins, such as vitamin D, in cats with IBD and ISCL is unknown.

Hypothesis/Objectives

The objective of this study was to measure serum 25 hydroxyvitamin D (25[OH]D) concentrations in cats with IBD or ISCL. Serum 25(OH)D also was measured in healthy cats, and in hospitalized ill cats with nongastrointestinal diseases.

Animals

Eighty‐four cats were included in the study: 23 in the healthy group, 41 in the hospitalized ill group, and 20 in the IBD/ISCL group.

Methods

Retrospective study. Serum samples for vitamin D analysis were frozen at −20°C until serum 25(OH)D was measured by high‐performance liquid chromatography (HPLC).

Results

Although there was overlap in serum 25(OH)D concentrations among the 3 groups, serum 25(OH)D concentrations were significantly lower in the cats with IBD or ISCL compared to healthy cats (P < .0001) and hospitalized ill cats (P = .014). In the IBD/ISCL group, there was a significant moderate positive correlation between serum albumin and 25(OH)D concentrations (r = 0.58, P = .018).

Conclusion and Clinical Importance

The median serum concentration of 25(OH)D was significantly lower in cats with IBD/ISCL than in healthy cats and in hospitalized ill cats. Additional studies are required to elucidate the mechanism of hypovitaminosis D in cats with gastrointestinal diseases, to define the best management strategy to treat this complication, and to investigate its potential prognostic implications.     

Establishment and Identification of a Fetal Fibroblast Cell Line with Double Transgenic (pGH/IGF-Ⅰ) Porcine

This study was aimed to establish a fetal fibroblast cell line of double transgenic (pGH/IGF-Ⅰ) pigs,and reserve fibroblast cells of double transgenic pigs for the follow-up study.A method of trypsin digestion was adopted to isolate and culture body tissues from pregnant porcine,and porcine fetal fibroblasts were isolated successfully through primary culture,subculturing and cryopreservation.The morphological observation,determination of viability before cryopreservation and after recovery,dynamic growth analysis,vimentin immunohistochemistry and microbial contamination detection were all done to study the biological characteristics of the cell line.The results showed that the fibroblasts were cultured and isolated successfully by trypsin digestion and differential centrifugation.The cell viability before cryopreservation and after recovery were 94.3% and 91.2%,respectively.The growth curve was sigmoidal,and experienced the incubation period,exponential growth period and platform three stages.The vimentin immunohistochemistry was positive,the microbial contamination detection were all negative.The results indicated that a fibroblast cell line of double transgenic porcine was successfully established.     

Flow Cytometric Characterization and Clinical Outcome of CD4+ T‐Cell Lymphoma in Dogs: 67 Cases

Background

Canine T‐cell lymphoma (TCL) is conventionally considered an aggressive disease, but some forms are histologically and clinically indolent. CD4 TCL is reported to be the most common subtype of TCL. We assessed flow cytometric characteristics, histologic features when available, and clinical outcomes of CD4+ TCL to determine if flow cytometry can be used to subclassify this group of lymphomas.

Objective

To test the hypothesis that canine CD4+ T‐cell lymphoma (TCL) is a homogeneous group of lymphomas with an aggressive clinical course.

Animals

Sixty‐seven dogs diagnosed with CD4+ TCL by flow cytometry and treated at 1 of 3 oncology referral clinics.

Methods

Retrospective multivariable analysis of outcome in canine CD4+ TCL including patient characteristics, treatment, and flow cytometric features.

Results

The majority of CD4+ TCL were CD45+, expressed low class II MHC, and exhibited an aggressive clinical course independent of treatment regimen (median survival, 159 days). Histologically, CD4+ TCL were classified as lymphoblastic or peripheral T cell. Size of the neoplastic lymphocytes had a modest effect on both PFI and survival in this group. A small number of CD4+ TCL were CD45− and class II MHC high, and exhibited an apparently more indolent clinical course (median survival not yet reached).

Conclusions and Clinical Importance

Although the majority of CD4+ TCL in dogs had uniform clinical and flow cytometric features and an aggressive clinical course, a subset had a unique immunophenotype that predicts significantly longer survival. This finding strengthens the utility of flow cytometry to aid in the stratification of canine lymphoma.     

Addition of Seminal Plasma to Post‐thawing Equine Semen: What is the Effect on Sperm Cell Viability?

Effect of seminal plasma addition after thawing on viability or cryocapacitation is not definitively established. This experiment was performed to verify the effect of adding seminal plasma, autologous or homologous (from an animal with good semen freezability). Five ejaculates from each of four stallions with proven fertility were collected and cryopreserved. The semen was subsequently thawed and divided into the following three treatment groups: no seminal plasma addition after semen thawing (NOSP); the addition of homologous seminal plasma after semen thawing (HSP) and the addition of autologous seminal plasma after semen thawing (ASP). The addition of 20% of seminal plasma led to an increase in the cell population that simultaneously show plasma and acrosomal membrane integrity (p < 0.05). The addition of seminal plasma did not alter the total motility, the amount of cells with mitochondrial membrane potential or the sperm velocities (average path velocity, straight-line velocity and curvilinear velocity). However, the beat/cross-frequency, straightness and linearity were reduced in ASP and HSP groups (p < 0.05). Unexpectedly, the addition of homologous seminal plasma reduced the proportion of cells with progressive motility (p < 0.05) and the addition of autologous seminal plasma reduced the amplitude of the lateral head displacement (p < 0.05). Based on the increase in the cell populations that had the plasma and acrosomal membrane integrity simultaneously identified in this study, we proposed that the addition of seminal plasma (autologous or homologous) into post-thawed semen before insemination could increase semen fertility.     

Generation of Human β-thalassemia Induced Pluripotent Stem Cells from Amniotic Fluid Cells Using a Single Excisable Lentiviral Stem Cell Cassette

Induced pluripotent stem cells (iPSCs) derived from somatic cells of patients represent a powerful tool for biomedical research and may have a wide range of applications in cell and gene therapy. However, the safety issues and the low efficiency associated with generating human iPSCs have limited their usage in clinical settings. The cell type used to create iPSCs can significantly influence the reprogramming efficiency and kinetics. Here, we show that amniotic fluid cells from the prenatal diagnosis of a β-thalassemia patient can be efficiently reprogrammed using a doxycycline (DOX)-inducible humanized version of the single lentiviral "stem cell cassette" vector flanked by loxP sites, which can be excised with Cre recombinase. We also demonstrated that the patient-derived iPSCs can be characterized based on the expression of pluripotency markers, and they can be differentiated into various somatic cell types in vitro and in vivo. Moreover, microarray analysis demonstrates a high correlation coefficient between human β-thalassemia iPS cells and human embryonic stem (hES) cells but a low correlation coefficient between human β-thalassemia amniotic fluid cells and human β-thalassemia iPS cells. Our data suggest that amniotic fluid cells may be an ideal human somatic cell resource for rapid and efficient generation of patient-specific iPS cells.     

Detection of Subclinical Mastitis in Dromedary Camels (Camelus dromedarius) using Somatic Cell Counts and the N-Acetyl-β-D-glucosaminidase Test

Somatic cell counts, N-acetyl--D-glucosaminidase (NAGase) activity and the infection status of the udder were determined in quarter milk samples (n = 86) from 22 multiparous, clinically healthy camels, traditionally managed by Bedouin nomads in the Negev desert, Israel. Seventy (81.4%) of the 86 samples examined contained bacteria, of which 35 (40.7%) gave mixed isolations of two or more bacteria, suggesting the existence of subclinical mastitis in the camel herds studied. Sixteen samples (18.6%) yielded no growth of bacteria. Staphylococcus aureus, Micrococcus spp., Bacillus spp., Streptococcus dysgalactiae and Escherichia coli were the main organisms isolated. The somatic cell count (SCC) ranged from 1.01×10⁵ to 11.78×10⁶ cells/ml. NAGase values were between 41.4 and 372 NAGase units. Quarter milk samples that contained bacteria had significantly (p<0.01) higher mean values for SCC but the mean NAGase levels were not significantly different for the bacteriologically negative and positive samples. There was a low correlation coefficient (r ² = 0.097) between the SCC and NAGase in the quarter milk samples from which bacteria were not isolated (n = 16) and a low negative correlation (r ² = –0.038) with the samples that contained bacteria (n = 70). The type of bacteria had a significant effect (p<0.01) on the SCC but not on the NAGase activity. Quarter samples from which Staphylococcus aureus (coagulase positive) was isolated showed the highest mean SCC and this organism is therefore suspected to be the underlying cause of the subclinical mastitis. The SCC gave a better indication of the presence of pathogenic microorganisms in milk samples than did NAGase.     

Effect of Duration of Packed Red Blood Cell Storage on Morbidity and Mortality in Dogs After Transfusion: 3,095 cases (2001–2010)

Background

Accumulating evidence suggests that transfusion of packed red blood cells (PRBCs) stored for >14 days is associated with increased rates of sepsis, multiple organ dysfunction, and mortality in human patients.

Objective

To determine if duration of PRBC storage has an effect on morbidity and mortality in dogs after transfusion.

Animals

Dogs admitted to the Matthew J Ryan Veterinary Hospital of the University of Pennsylvania.

Methods

A retrospective case review of dogs identified through blood bank logbooks that received PRBC transfusions (minimum, 5 mL/kg) between 2001 and 2010. Dogs were categorized according to major cause of anemia (eg, hemorrhage, hemolysis, ineffective erythropoiesis) for analysis.

Results

A total of 3,095 dogs received 5,412 PRBC units. Longer duration of PRBC storage was associated with development of new or progressive coagulation failure (P = .001) and thromboembolic disease (P = .005). There was no association between duration of PRBC storage and survival for all dogs overall. However, a logistic regression model indicated that for dogs with hemolysis, 90% of which had immune‐mediated hemolytic anemia, longer duration of PRBC storage was a negative risk factor for survival. For every 7 day increase in storage, there was a 0.79 lesser odds of 30 day survival (95% CI, 0.64–0.97; P = .024).

Conclusions and Clinical Importance

Duration of PRBC storage does not appear to be a major contributing factor to mortality in the overall canine population. However, longer duration of PRBC storage may negatively impact outcome in dogs with immune‐mediated hemolytic anemia, thus warranting further investigation with prospective studies.     

Effects of Compound Preparation of Chinese Herbal Extract on Performance,Immune and Antioxidant Function and Ruminal Bacterial Flora of Cows with High Somatic Cell Count In Milk北大核心CSCD

The aim of this experiment was to investigate the effects of compound preparation of Chinese herbal extract on performance, immune and antioxidant function and ruminal bacterial flora of cows with high somatic cell count (SCC) in milk. Ten transitional Holstein cows with similar body condition, lactation period, milk yield and milk SCC higher than 500 000 cells / mL were selected and divided into control group [milk SCC = (97.48±23.75) ten thousand cells / mL] and experimental group [milk SCC = (98. 78 ± 29. 92) ten thousand cells / mL] based on the principle of similar milk SCC, each group contained 5 cows, and cows in 2 groups were not treated with medication. The control group was fed a total mixed ration (TMR), and the experimental was fed the TMR+54 g / (d.head) compound preparation of Chinese herbal extract. The experiment lasted for 35 days. The results showed as follows: 1) compared with the control group, the addition of compound preparation of Chinese herbal extract in the diet significantly decreased the SCC in milk at days 14 and 28 of cows with high SCC in milk (P<0.05), but had no significant effects on milk yield, milk composition and nutrient apparent digestibility (P>0.05) . 2) Compared with the control group, the addition of compound preparation of Chinese herbal extract in the diet significantly increased the contents of immunoglobulin A (IgA), immunoglobulin M (IgM) and interferon-γ (IFN-γ) in serum of cows with high SCC in milk (P<0.05), and significantly decreased the serum tumor necrosis factor-α (TNF-α) content (P<0.05) . 3) Compared with the control group, the addition of compound preparation of Chinese herbal extract in the diet significantly decreased the proportions of Actinobacteria, Succinivibrionaceae_UCG-001 and Ruminococcus_gauvreauii_group in rumen of cows with high SCC in milk (P < 0. 05), and significantly increased the rumen Prevotellaceae_UCG-001 proportion (P<0.05) . It is concluded that the addition of compound preparation of Chinese herbal extract in the diet can reduce the SCC in milk, increase the immune function, and improve the rumen bacterial flora composition of cows with high SCC in milk. © 2023 Authors. All rights reserved.     

A Review of Current Treatment Options in the Treatment of Ocular and/or Periocular Squamous Cell Carcinoma in Horses: Is There a Definitive “Best” Practice?

This review examines the most commonly reported treatment options for ocular squamous cell carcinoma (OSCC) and periocular squamous cell carcinoma (POSCC) in horses and proposes to conclude on the most viable method based on available published studies in terms of treatment outcome, known side effects, advantages, disadvantages, and reliability of available evidence. After a literature search for peer-reviewed published articles, seven most commonly reported on treatments for OSCC and/or POSCC were identified: surgery, photodynamic therapy, carbon dioxide (CO₂) laser ablation, radiofrequency hyperthermia, cryotherapy, chemotherapy, and radiation therapy. Combination therapies were supported as a most successful recommendation; however, when considering site-specific outcomes, the following conclusions may be drawn: limbal squamous cell carcinoma (SCC) was most effectively treated with surgery and adjunctive therapy including CO₂ laser ablation, mitomycin C, and brachytherapy; third eyelid SCC reported good outcomes when treated with surgery alone (clear margins) and in combination with brachytherapy for unclear margins; eyelid SCC, surgical resection was usually limited and most reports supported the use of adjunctive brachytherapy, although photodynamic therapy appeared to be a promising new treatment. It was deemed unreasonable to conclude on the best treatment for cornea, conjunctivae (palpebral and bulbar), and medial canthi in isolation because of lack of evidence. A consistently favored treatment for OSCC and/or POSCC in horses does not currently exist. The presentation of data in the literature and its lack of consistency make it impossible to statistically analyze and make comparative conclusions on treatment outcomes. This review provides a basis for further research to establish a best-practice protocol.