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Objective: PD-1/PD-L1 immune checkpoint treatment is effective for some triple-negative breast cancer populations with PD-L1 expression, but the response rate is still not satisfactory. This study aims to explore the mechanism of drug resistance to breast cancer anti-PD-1 therapies and the strategies for overcoming the resistance to PD-1therapies. Methods: By constructing a human triple-negative breast cancer drug-resistant cell line called BT-549R5 and a mouse breast cancer drug-resistant cell line called 4T1R3, and applying the whole-gene shRNA library screening, candidate drug resistance-associated molecules were obtained and verified by cytological experiments. The expression of Tyro3, Axl and MerTK of the TAM family in the 4T1R3 group was tested using the Western blot method. The down-regulation of CDK9 on the effect of T cells killing the BT-549R5 cells was observed through T cell killing tests, while the down-regulation of Tyro3 and CDK9 on the effect of anti-PD-1 therapies for transplanted breast tumors was observed in mouse tumor formation experiments. Results: The cell lines and animal models of breast cancer resistant to PD-1 treatment were successfully constructed. Tyro3, Axl and MerTK were highly expressed in 4T1R3 cells. Whole genome sequencing showed that Tyro3 and CDK9 were highly expressed in BT-549R5 cells. T cell killing experiment showed that the survival rate of BT-549R5 cells in the CDK9 down-regulated group and the control group decreased gradually with the increase of T cells, but the survival rate of BT-549R5 cells in the CDK9 down-regulated group decreased rapidly. Tumor formation experiment in mice showed that under anti-PD-1 treatment, the transplanted tumor in the 4T1R3 cell group grew rapidly compared with the 4T1 cell group (P<0.05), and the tumor volume of the 4T1R3 group was larger than that of the 4T1 group on Day 20. Nevertheless, the tumor growth rates in the CDK9-knockdown 4T1R3 cell group and the Tyro3-knockdown 4T1R3 cell group were similar to that of the 4T1 cell group, and the tumor volumes at day 20 were signiference lower than that of 4T1R3 cell group(P<0.05). Conclusions: Tyro3 and CDK9 are associated with the drug resistance to anti-PD-1 therapies for breast cancer. Inhibiting the expression of Tyro3 and CDK9 can reverse the drug resistance to breast cancer treatment.
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Humanos , Animais , Camundongos , c-Mer Tirosina Quinase/metabolismo , Receptores Proteína Tirosina Quinases/genética , Receptor Tirosina Quinase Axl , Proteínas Proto-Oncogênicas/metabolismo , Antígeno B7-H1/genética , Neoplasias de Mama Triplo Negativas/genética , Resistencia a Medicamentos Antineoplásicos , Biomarcadores , Linhagem Celular Tumoral , Quinase 9 Dependente de CiclinaRESUMO
Objective: PD-1/PD-L1 immune checkpoint treatment is effective for some triple-negative breast cancer populations with PD-L1 expression, but the response rate is still not satisfactory. This study aims to explore the mechanism of drug resistance to breast cancer anti-PD-1 therapies and the strategies for overcoming the resistance to PD-1therapies. Methods: By constructing a human triple-negative breast cancer drug-resistant cell line called BT-549R5 and a mouse breast cancer drug-resistant cell line called 4T1R3, and applying the whole-gene shRNA library screening, candidate drug resistance-associated molecules were obtained and verified by cytological experiments. The expression of Tyro3, Axl and MerTK of the TAM family in the 4T1R3 group was tested using the Western blot method. The down-regulation of CDK9 on the effect of T cells killing the BT-549R5 cells was observed through T cell killing tests, while the down-regulation of Tyro3 and CDK9 on the effect of anti-PD-1 therapies for transplanted breast tumors was observed in mouse tumor formation experiments. Results: The cell lines and animal models of breast cancer resistant to PD-1 treatment were successfully constructed. Tyro3, Axl and MerTK were highly expressed in 4T1R3 cells. Whole genome sequencing showed that Tyro3 and CDK9 were highly expressed in BT-549R5 cells. T cell killing experiment showed that the survival rate of BT-549R5 cells in the CDK9 down-regulated group and the control group decreased gradually with the increase of T cells, but the survival rate of BT-549R5 cells in the CDK9 down-regulated group decreased rapidly. Tumor formation experiment in mice showed that under anti-PD-1 treatment, the transplanted tumor in the 4T1R3 cell group grew rapidly compared with the 4T1 cell group (P<0.05), and the tumor volume of the 4T1R3 group was larger than that of the 4T1 group on Day 20. Nevertheless, the tumor growth rates in the CDK9-knockdown 4T1R3 cell group and the Tyro3-knockdown 4T1R3 cell group were similar to that of the 4T1 cell group, and the tumor volumes at day 20 were signiference lower than that of 4T1R3 cell group(P<0.05). Conclusions: Tyro3 and CDK9 are associated with the drug resistance to anti-PD-1 therapies for breast cancer. Inhibiting the expression of Tyro3 and CDK9 can reverse the drug resistance to breast cancer treatment.
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Humanos , Animais , Camundongos , c-Mer Tirosina Quinase/metabolismo , Receptores Proteína Tirosina Quinases/genética , Receptor Tirosina Quinase Axl , Proteínas Proto-Oncogênicas/metabolismo , Antígeno B7-H1/genética , Neoplasias de Mama Triplo Negativas/genética , Resistencia a Medicamentos Antineoplásicos , Biomarcadores , Linhagem Celular Tumoral , Quinase 9 Dependente de CiclinaRESUMO
The basic knowledge of college chemistry is an important basis for the key undergraduate course-Biochemistry for students majoring in Life Sciences. Based on theoretical analysis of the close relationship between "Biochemistry" and "College Chemistry", this paper summarized these knowledge nodes to the content of Biochemistry from the four major courses: Chemistry, Inorganic Chemistry, Organic Chemistry, Analytical Chemistry and Physical Chemistry. We took the peptide bond,mechanism of enzymes and protein purification as examples to show the relevance between the basic knowledge of Chemistry and the knowledge points of Biochemistry, aiming to enlighten the students to understand the importance of the basic knowledge of Chemistry. We also proposed three kinds of teaching pattern for introducing the basic knowledge of " College Chemistry" to " Biochemistry" , including " Described directly by teachers", " Integrated into contents of Biochemistry" and " Students learning by themselves" , so as to ensure that the students comprehend clearly the essence of "Biochemistry". Finally we discussed how to culture the students' ability to think independently and critically. The idea and practice about Biochemistry teaching will benefit the students to grasp the essence of the course and it will also give one new perspective for teachers to carry out teaching reforms in life science courses.
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<p><b>OBJECTIVE</b>To study the expression of transforming growth factor-β1 (TGF-β1) and plasminogen activator inhibitor-1 (PAI-1) and its significance in premature infants with bronchopulmonary dysplasia (BPD).</p><p><b>METHODS</b>A retrospective analysis was performed on the clinical data of 96 very low birth weight infants (gestational age of ≤ 32 weeks) who survived for more than 28 days and were admitted to the Neonatal Intensive Care Unit between January 2010 and December 2012. These subjects were divided into BPD group (n=21) and non-BPD group (n=75). The expression of TGF-β1 and PAI-1 in blood was measured by ELISA.</p><p><b>RESULTS</b>The levels of TGF-β1 and PAI-1 in the BPD group increased gradually from the 7th day to the 14th day and then to the 21st day after birth, and were significantly higher than in the non-BPD group at all time points (P<0.01). The TGF-β1 and PAI-1 levels in the non-BPD group on the 7th, 14th, and 21st days after birth were not significantly different from each other (P>0.05).</p><p><b>CONCLUSIONS</b>The expression of TGF-β1 and PAI-1 in blood is elevated in premature infants with BPD, which may be associated with the development of BPD.</p>
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Feminino , Humanos , Recém-Nascido , Masculino , Displasia Broncopulmonar , Metabolismo , Ensaio de Imunoadsorção Enzimática , Recém-Nascido Prematuro , Inibidor 1 de Ativador de Plasminogênio , Sangue , Estudos Retrospectivos , Fator de Crescimento Transformador beta1 , SangueRESUMO
Objective To investigate the influence of surgery with the guideline of minimally invasive concept in prognosis of patients with hypertensive basal ganglia hematomas. Methods Fifty-seven patients with hypertensive intra-cerebral hemorrhage were randomly divided into 2 groups:Group A (admitted to our hospital from January 2007 to December 2008 and performed surgery under the condition that the content of hematoma reached the level for surgery,n=26) and Group B (admitted to our hospital from January 2009 to June 2011 and received surgery with the guideline of minimally invasive concept once noting the tendency ofexpanded hematoma,n=31).We evaluated the influence of surgery (total removal of the hematoma and proper stopping the bleeding) according to the condition that tendency of expanded hematoma appeared and with the guideline of minimally invasive concept in the prognosis of these patients. Results No significant differences in consciousness classification and hematoma volume before surgery were noted between the 2 groups (P>0.05).Responsible vessels were noted in 15 patients from Group A and 27 patients from Group B, and significant difference was noted between these 2 groups (P<0.05).The hematoma clearance rate was 75% in Group A,and re-bleeding was noted in 4 patients (15.4%) after the surgery; while that was higher than 90% in Group B, and re-bleeding was only noted in 2 patients (6.5%) whose responsible vessels could not be found.The good recovery rate in Group A was 46.2% 3 months after surgery, while that in Group B was 74.2%, which indicated that the effect in group B was obviously better than that in group A (P<0.05). Conclusion Tendency of expanded hematoma should be paid attention in patients with hypertensive basal ganglia cerebral hemorrhage; it is important to quickly identify the cases showing clear indications for surgery and to perform the procedures at the earliest time; the procedures, including completely removal of the hematoma and properly stopping the stanch bleeding under direct vision with the guideline of minimally invasive concept can improve the recovery fiom hypertensive basal ganglia cerebral hemorrhage.
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<p><b>OBJECTIVE</b>To study the pathological changes in the non-myelin sheath by observing histological damages to the neurofilament protein and apoptosis of neurons in rats with experimental autoimmune encephalomyelitis (EAE).</p><p><b>METHODS</b>Forty-eight Wistar rats were randomly divided into two groups: control and EAE (24 rats in each group). Behavioral changes were observed. Inflammation reactions and demyelination were observed by hematoxylin eosin staining and LOYEZ staining.The level of neurofilament was detected by immunohistochemistry. Apoptosis of the neuron in the spinal cord was detected by TUNEL.</p><p><b>RESULTS</b>Behavioral and histological results confirmed that the model of EAE rats was prepared successfully. In the EAE group, typical morphological features of axonal damage (sparsed axonal density, axonal distortion, axonal transection and even axonal disappearance) were found from the seventh day after immunization and the morphological changes were the most obvious on the fourteenth day. Neurofilament density in the EAE group was significantly lower than in the control group (P<0.01) at 7, 14 and 21 days after immunization. The neuronal apoptosis index in the EAE group at 7, 14 and 21 days after immunization was significantly higher than in the control group (P<0.01).</p><p><b>CONCLUSIONS</b>In addition to inflammatory demyelination, axonal damage and neuronal apoptosis can be observed in the early stage of EAE. Pathological changes may be associated with neurological dysfunction.</p>
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Animais , Feminino , Ratos , Apoptose , Axônios , Patologia , Encefalomielite Autoimune Experimental , Patologia , Psicologia , Imuno-Histoquímica , Bainha de Mielina , Patologia , Proteínas de Neurofilamentos , Neurônios , Patologia , Ratos Wistar , Medula Espinal , PatologiaRESUMO
<p><b>OBJECTIVE</b>To explore the brain activation before and in early period after olfactory adaptation using functional magnetic resonance imaging, and discuss the mechanisms of olfactory adaptation.</p><p><b>METHODS</b>Ten right-handed, normosmic subjects underwent 2 times of olfactory stimulation tasks with the interval of 20 minutes. The odorant used was isovaleric acid. The fMRI data was processed by the SPM5 software. Rating odor intensity and valence using visual analogue scale (VAS), and the results of 2 tasks were statistically analyzed.</p><p><b>RESULTS</b>There was no significant difference between 2 tasks on both intensity and hedonicity scores. In task 1, the brain activation in bilateral cerebellum, frontal (including orbitofrontal gyrus), insula, thalamus, cingulate gyrus, putamen, amygdala, piriform cortex, the left inferior parietal lobule, precentral gyrus, right hippocampus, pallidum, middle temporal gyrus, supramarginal gyrus. In task 2, only the right middle frontal gyrus activated, and the voxels decreased significantly. Paired t-test results showed that: (task1-task2) activated regions in left precentral gyrus, frontal lobe (including the orbitofrontal gyrus), insula, right superior temporal gyrus, cerebellum; (task2-task1) activation in the left inferior parietal lobule and right lingual gyrus.</p><p><b>CONCLUSIONS</b>The sensitivity of brain activation is still at a low level, when subjects had recovered from adaptation in subjective olfactory perception. Underwent repeated olfactory stimulation, second olfactory cortex plays less role on olfactory perception and advanced processing.</p>
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Adulto , Feminino , Humanos , Masculino , Adaptação Fisiológica , Córtex Cerebral , Fisiologia , Imageamento por Ressonância Magnética , Métodos , Percepção Olfatória , FisiologiaRESUMO
<p><b>OBJECTIVE</b>To observe whether injectable hydrogel implantation could prevent adverse cardiac remodeling and preserve cardiac function in rabbits with myocardial infarction (MI).</p><p><b>METHODS</b>A novel injectable hydrogel, the copolymer MPM/alpha-CD, which self-assembled between alpha-cyclodextrin and methoxy polyethylene glycol-poly (caprolactone)-(dodecanedioic acid)-poly (caprolactone)-methoxypolyethylene glycol triblock polymer, was synthesized by chemical crosslinking and characterized by biocompatibility and biodegradability. Experimental MI was induced in male rabbits by coronary artery ligation. The MI rabbits were randomly divided into hydrogel group (200 microl MPM/alpha-CD were injected into the infarcted myocardium 7 days after MI) and control group (equal volume phosphate buffered saline myocardial injection, n = 8 each). Four weeks after MPM/alpha-CD implantation, echocardiography, histochemistry and immunohistochemistry were performed.</p><p><b>RESULTS</b>Left ventricle ejection fraction was significantly improved in the hydrogel-treated group (56.1% +/- 8.4%) than that in the control group (37.3% +/- 6.4%, P < 0.05). Histological analysis indicated that hydrogel degraded 4 weeks after hydrogel injection, and prevented scar expansion and wall thinning [(3.08 +/- 0.32) mm vs. (2.18 +/- 0.46) mm in control group, P < 0.05]. Neovasculature formation was similar between the hydrogel group [(100.8 +/- 2.4)/mm(2)] and the control [(98.5 +/- 2.9)/mm(2), P > 0.05].</p><p><b>CONCLUSION</b>MPM/alpha-CD could serve as an excellent injectable biomaterial for improves cardiac function and attenuating scar expansion and left ventricular dilation in MI rabbits.</p>
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Animais , Masculino , Coelhos , Ventrículos do Coração , Hidrogel de Polietilenoglicol-Dimetacrilato , Farmacologia , Usos Terapêuticos , Infarto do Miocárdio , Terapêutica , Remodelação VentricularRESUMO
Primary renal lymphoma is one of the malignant lymphomas that initially presents in the extra lymphonode, which is rarely seen in children. This study reported two cases of primary renal lymphoma in children who were definitively diagnosed by renal biopsy. Renal tubular acidosis was the initial manifestation in both cases. They were referred to the hospital with chief complaints of polydipsia, polyuria, debilitation, vomiting and anemia. Imaging and laboratory examinations showed bilateral renomegaly, hypocalcemia, hypophosphatemia, and metabolic acidosis. One of the patients discontinued therapy. The other received chemotherapy including prednisone, vincristine, cytarabine and L-asparaginase, combined with intrathecal injections of methotrexate, dexamethasone and Ara-C and supporting treatment. Twenty-three days after treatment, polydipsia and polyuria were relieved, and acidosis, kaliopenia and anemia were improved in the patient. There were no abnormal findings in the renal B-ultrasound re-examination. It was concluded that when a patient is suspected of renal lymphoma, diagnostic puncture and renal biopsy should be performed early. Early combined therapeutics including chemotherapy, radiation therapy, surgery and supporting treatments may result in a favorable prognosis in patients with this disease.
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Criança , Humanos , Masculino , Acidose Tubular Renal , Diagnóstico , Neoplasias Renais , LinfomaRESUMO
<p><b>OBJECTIVE</b>To explore the diagnosis and treatment of penile verrucous carcinoma.</p><p><b>METHODS</b>The clinical and pathological data of 4 patients with penile verrucous carcinoma were analyzed.</p><p><b>RESULTS</b>The patients ranged in age from 42 to 76 years (average 52). All the tumors showed exophytic papillary lesions, the biggest being 2.1 to 5.8 cm in diameter. The lesions were confined to the glans penis in two cases and invaded the shafts in the other 2 (1 accompanied by syphilis). One patient, whose tumor was small (1.4 cm in diameter) and confined to the glans penis, underwent glandectomy. One with a larger tumor confined to the glans penis and the other 2 with the shafts involved underwent partial penectomy, including the one accompanied by syphilis, who underwent the operation after treated by Benzathine benzylpenicillin. Histopathological examination of the specimens showed that the tumor cells were mostly well-differentiated and the surgical margins were tumor free in all the 4 cases. HE stain was performed in all the specimens. Microscopic examination revealed papillomatosis and hyperkeratosis of the epithelium, with bulbous projections into the lamina propria consisting of well-differentiated squamous epithelial cells. Marked invasion of the stroma by lymphocytes was noted. Follow-up ranged from 3 to 7 years (average 4.6), revealing no recurrence. The result of the rapid plasma regain (RPR) test was negative but that of the Treponema pallidum passive-particle-agglutination (TPPA) test remained positive in the blood of the patient accompanied by syphilis after treatment.</p><p><b>CONCLUSION</b>Verrucous carcinoma of the penis is characterized by low malignant potential and locally aggressive nature. It seldom develops metastasis to regional lymphonodes or distant areas. Glandectomy or partial penectomy can be chosen for its treatment, with favorable prognosis.</p>
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Idoso , Humanos , Masculino , Pessoa de Meia-Idade , Carcinoma Verrucoso , Diagnóstico , Patologia , Cirurgia Geral , Seguimentos , Neoplasias Penianas , Diagnóstico , Patologia , Cirurgia GeralRESUMO
<p><b>OBJECTIVE</b>To study the effects of heat shock preconditioning on the expression of heat shock protein-70 (HSP70) and apoptosis of the neuron in experimental autoimmune encephalomyelitis (EAE) rats.</p><p><b>METHODS</b>Thirty-six Wistar rats were randomly divided into control, EAE and heat shock preconditioning groups (n=12 each). The EAE animal model was induced with guinea pig myelin basic protein. Heat shock preconditioning was performed 24 hrs prior to the EAE model inducement. No treatment was done in the control group. The neurological signs were observed after immunization. The spinal cords were removed and stained with hematoxylin and eosin. HSP70 was detected by immunohistochemistry. Apoptosis of the neuron was measured by TUNEL.</p><p><b>RESULTS</b>Heat shock preconditioning significantly alleviated clinical signs and neuronal injury. HSP70 expression in the heat shock preconditioning group was significantly higher than in the untreated EAE group (21.08 +/- 0.87 vs 10.17 +/- 0.51; P < 0.01). Heat shock preconditioning suppressed apoptosis of the neuron compared with the EAE group (apoptosis rate: 21.92 +/- 1.00% vs 58.92 +/- 1.67%; P < 0.01).</p><p><b>CONCLUSIONS</b>Heat shock preconditioning might improve the neurological outcome in EAE rats, possibly through the induction of HSP70 synthesis and the reduction of apoptosis of the neuron in spinal cords.</p>
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Animais , Feminino , Masculino , Ratos , Apoptose , Medula Óssea , Patologia , Encefalomielite Autoimune Experimental , Patologia , Terapêutica , Cobaias , Proteínas de Choque Térmico HSP70 , Temperatura Alta , Neurônios , Patologia , Ratos WistarRESUMO
<p><b>OBJECTIVE</b>To determine the threshold of millimeter wave irradiation for fetal injury in mice and the mechanism of decrease of learning and memory function in their offsprings and to verify whether the millimeter wave has the non-thermal effect.</p><p><b>METHODS</b>Pregnant mice were irradiated by millimeter wave with frequencies of 37.4, 42.2, 53.0 and 60.0 GHz at power densities of 1, 3, 5, 8 mW/cm(2) for two hours daily from the 6th to 15th day of their gestation. Learning and memory functions of their offsprings were tested by a Y-type electric maze. c-Fos protein expression level in hippocampus of their offsprings was determined with immunohistochemistry 0, 30, 60, 90 and 120 minutes after the offsprings were trained respectively.</p><p><b>RESULTS</b>The minimal power density of millimeter wave for the decrease in learning and memory function and decrease of c-Fos protein expression level in hippocampus of their offsprings caused by 37.4, 42.2 GHz and 53.0, 60.0 GHz was 5 and 3 mW/cm(2). Severity of injury for learning and memory in offsprings caused by irradiation increased with the power density of millimeter wave. The millimeter wave did not cause increase of the body temperature of the pregnant mice.</p><p><b>CONCLUSION</b>The threshold of millimeter wave with 37.4, 42.2 GHz, and 53.0, 60.0 GHz causing fetal injury in mice is 5 and 3 mW/cm(2) respectively. The decrease in learning and memory functions in offspring mice is related with decrease of c-Fos protein expression level in hippocampus. Millimeter wave has the non-thermal effects.</p>
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Animais , Feminino , Camundongos , Gravidez , Relação Dose-Resposta à Radiação , Hipocampo , Metabolismo , Efeitos da Radiação , Imuno-Histoquímica , Aprendizagem , Efeitos da Radiação , Memória , Efeitos da Radiação , Camundongos Endogâmicos BALB C , Micro-Ondas , Efeitos Tardios da Exposição Pré-Natal , Proteínas Proto-Oncogênicas c-fosRESUMO
Objective To study the clinical characteristics of children with demyelinating disease.Methods Age of onset,presymptoms,clinical manifestations and auxiliary examinations of 51 children diagnosed as demyelinating disease were analyzed retrospectively.Results The onset age was from 3 months to 14 years,and the number of school age children was 38.Before illness,32 cases were relevant to infection,3 cases to vaccine inoculation.Most of children had acute courses.In the initial stage,32 cases with peripheral nervous demyelinating disease presented to paralysis of limbs;18 cases with central nervous demyelinating disease presented to visual disorder,somatasthenia,fever,convulsion and headache.The EMGs of children with peripheral nervous demyelinating disease showed nervous lesion.Among 18 children with central nervous demyelinating disease,17 cases showed abnomal signal on MRI or CT.Conclusions Children with demyelinating disease displays diversified clinical manifestations.By investigating case history combined with auxiliary exaninations,it is not difficult to make a correct diagnosis and its prognosis is good commonly.