Objective: To study the effect of berberine (BR) on ventricular remodeling in experimental rats with myocardial infarction (MI) and its mechanisms.
Methods: The MI model of experimental rats was established by ligation of the left anterior descending coronary artery and the MI animals were randomly divided into 3 groups: MI+BR group, in which the rats received BR 20 mg/kg.d, Sham group and MI group, the rats in those 2 groups received the same volume of normal saline. All animals were treated for 8 weeks. The cardiac function and structure were assessed by echocardiography, cardiac interstitial collagen deposition was evaluated by Masson stain, the myocardial cell apoptosis was detected by Tunel method, and the activation of nuclear factor (NF-κB) was also examined.
Results: For echocardiography, MI group had enlarged left ventricular end diastolic diameter (7.28 ± 0.29) mm than Sham group (6.86 ± 0.36) mm,P<0.05, but it decreased in MI+BR group (6.89 ± 0.99) mm,P>0.05. MI group had increased left ventricular end systolic diameter (5.88 ± 0.33) mm than Sham group (4.61 ± 0.31) mm, but it decreased in MI+BR group (4.68 ± 1.17) mm, allP< 0.01. MI group showed increased left ventricular posterior wall compensatory hypertrophy (1.81 ± 0.85) mm than Sham group (1.67 ± 0.16 mm),P<0.05, while in MI+BR group, it was deereased to (1.65 ± 0.14) mm. MI group presented decreased LVEF (45.77 ± 3.17) % than Sham group (67.28 ± 4.15) %, but it increased in MI+BR group (64.64 ± 5.82) %, allP<0.01. For Masson stain, cardiac interstitial collagen deposition in MI group (11.39 ± 0.45) % was higher than Sham group (2.65 ± 0.45) %, but less in MI+BR group (7.00 ± 0.87) %, allP<0.01. For Tunel examination, the myocardial cell apoptosis index was increased in MI group (21.31 ± 2.34) than Sham group (0.99 ± 0.38), but decreased in MI+BR group (14.15 ± 1.62), allP<0.01. For NF-κB activation study, the nuclear protein p65 content was higher in MI group (0.14 ± 0.02) ng/ml than Sham group (0.06 ± 0.01) ng/ml, but lower in MI+BR group (0.10 ± 0.02) ng/ml, allP<0.01.
Conclusion: Application of BR may improve the ventricular remodeling and cardiac function in experimental MI rats, it might be because of BR partially inhibit NF-κB activation, reduce collagen deposition and help anti-apoptosis in myocardial cells.

Objectives Tostudytheexpressionchangesofproproteinconvertase1(PC1)incerebral cortex nerve cells and its substrate neuropeptide Y (NPY)after focal cerebral ischemia in mice and to investigatetheeffectofPC1inneuronalischemicinjury.Methods Twenty-fourmaleC57micewere randomly allocated into a sham-operation group,an ischemia-reperfusion 4-or 24-hour group with computer (n=8 in each group). A rat model of middle cerebral artery occlusion was induced by the intraluminal suture method. Western blot and real-time quantitative nucleic acid amplification were used to detect the expression changes of PC1,NPY,and mRNA in mouse cortical neurons. Results (1)Compared with the sham operation group,the expression of PC1 mRNA of ischemic cortex brain tissue at ischemic side in the ischemia-reperfusion 4-hour group increased 2. 66 ± 0. 24 and in the ischemia-reperfusion 24-hour group expressed 2. 07 ± 0. 23 (all P<0. 05). Compared with the sham operation group,the PC1 precursor protein level increased significantly at 4 hours (P<0. 05). There was no significant difference in the 24-hour group (P >0. 05 ). (2 )Compared with the sham operation group,the preproNPY mRNA and protein level increased significantly after reperfusion in the ischemia-reperfusion 4-hour group (P < 0. 05 ),the mRNA expressed 2. 31 ± 0. 27,and the increase of precursor protein level continued until 24 hours. Conclusion TheexpressionofprecursorPC1increasedaftercerebralischemia-reperfusioninmice, thus affecting the processing activity of PC1 ,and resulting in NPY protein,an active substrate of PC1 accumulated with the form of precursors,which may be one of the underlying mechanisms of neuronal ischemic injury.

Objective To study the prognostic and predictive significance of 21-gene assay ( oncotype DX)in breast cancer.Methods Real-time quantitative PCR(RT-QPCR)was used to detect 21 gene expression in breast cancer tissues (100 cases)and recurrence score(RS)was calculated.Results Among the 100 cases, 52 cases had low RS , 22 cases had middle RS , and 26 cases had high RS .The recurrence rate of five years was 1.92%,4.55%and 15.38%respectively.21 gene expression had nothing to do with patients'age, tumor size, histological grade , lymph node metastasis state , ER expression , or PR expression .It was associated with HER 2 expression .Conclusions 21 genes is a good prediction factor in breast cancer and its prognosis .

Objective To investigate ERCC1 expression in advanced breast cancer and its relationship with cisplatin resistance. Methods ERCC1 expression in 50 cases of advanced breast cancer was measured by RT-QPCR. The chemotherapy (CAF) was given. Gemcitabine and cisplatin were given because of metastasis. Results The expression of ERCC1 has no relationship with age, lymph node metastasis, clinicl stage, histologi-cal grade or human epidermal growth factor receptor-2 (HER2) expression. The effective rate was 18.8% (3/16) for ERCC1 high-expressed patients and 79.4%(27/34)for ERCC1 low-expressed patients in terms of cisplatin chemotherapy. The sensitity for cisplatin chemotherapy was high for patients with low ERCC1 expression and it was low for patients with high ERCC1 expression. The effective rate (complete remission+partial remission), and ineffective rate (stable disease+progressive disease) between the two had statastical significance (P<0.001). Con-clusions ERCC1 low-expressed patients can benefit from cisplatin. ERCC1 can be used as a moleculer marker for predicting chemotherapy efficacy in breast cancer.

Objective To investigate the expression of thymidylate synthase (TS) in breast cancer and its relationship with fluorouracil sensibility and patients' prognosis.Methods TS expression in 80 cases of breast cancer was measured by RT-QPCR.The chemotherapy (CAF) was given.Relationship between TS and fluorouracil sensibility was studied,as well as the relation between TS expression and prognosis of breast cancer.Result Positive expression of TS gene was detected by Q-PCR 1n 27.5％ of the breast cancer specimens.The expression of TS had no relation with age,lymphatic metastasis,histological grade or clinical stage,while it was obviously correlated with HER2 (P＜0.01).The expression of TS had negative correlation with the antitumor effect of fluorouracil.The five-year survival rate of TS positive patients was lower than that of TS negative patients (P＜0.01).Conclusions Breast cancer patients with low expression of TS may benefit from fluorouracil.TS can be used as a molecule marker for predicting efficacy of breast cancer chemotherapy.

Objective:To investigate characteristic CT manifestations of primary pulmonary diffuse large B-cell lymphoma (DLBCL).Methods:CT images of 14 patients [10 males and 4 females, age （54±2） years old, range 32 to 91] with pathologically-proved pulmonary DLBCL lymphoma were retrospectively analyzed．Plain CT and contrast enhanced CT imaging were performed in all 14 patients. Image characteristics including lesion size, locations and distribution, morphology and margin, density and enhancement degrees, bronchia and lesion surroundings, other thoracic extra-pulmonary manifestations, as well as distant metastasis were analyzed and recorded. The maximal diameter of mass and/or nodules, pre and post-contrast CT values were measured. Among all 14 cases, 8 cases were initially diagnosed as lung carcinoma, 5 cases as infection, one case as lymphoma.Results:Among all 14 primary lung DLBCL cases, there were 10 case with multiple lesions and 4 with single lesion. Masses and/or nodules were found in 12 cases, with the maximum diameter of the lesions as 0.8-8.2 cm, the median value as 5.3 (2.9, 7.8) cm. Two cases showed simple consolidation. The margins of the lesions were clear and smooth in 12 cases, and fuzzy in 2 cases. The density of the lesions on pre-contrast CT was relatively uniform, with mean CT value (35.1±1.0) HU. After contrast, 10 cases displayed mild to moderate homogeneous enhancement, 4 cases showed heterogenous enhancement. The mean CT value of post-contrast images was (61.8±1.5) HU. In arterial phase, the mean CT value was (50.9±1.3) HU. Angiographic sign was found in 9 cases in arterial phase. Of the 14 cases, bronchus was clear and smooth in 5 cases. In 4 cases, bronchus was found slight compressed or stenosis; and 5 cases showed intra-lesion bronchi invasion or occlusion. Interstitial tissue around the lesion was found slightly thickened in 8 cases. The pleura showed unevenly thickened and invaded in 8 cases. Mediastinal or hilar lymphadenopathy and fusion were found in 10 cases, with 3 cases involving mediastinal large blood vessels, and 7 cases displaying infiltrative growth pattern. There were 4 cases with pleural effusion. CT follow-up after treatment in 8 cases showed no distant metastasis (7 cases showed good prognosis, with lesions disappearing after radiotherapy, chemotherapy or surgical resection; 1 case showed progressed with lesion increased after chemotherapy). Six patients abandoned the treatment and discharged from the hospital.Conclusions:Primary DLBCL is a high invasive and malignant entity with certain CT characteristics. The confirmed diagnosis of pulmonary DLBCL depends on pathological results.

Objective To observe the dynamic changes of cardiac lymphangiogenesis in Doxorubicin(DOX)-induced dilated cardiomyopathy(DCM)model mice,and to study the the protective mechanism of Kuoxin Decoction.Methods The DCM mouse model was established by intraperitoneal injection of DOX,and the dynamic observation was performed every week.On this basis,60 C57BL/6 mice were randomly divided into 6 groups(n=10):control group,Model group,L-KXD,M-KXD and H-KXD groups and Captopril group.After successful modeling,the KXD and the positive control drug Captopril were administered continuously for 28 days.Echocardiography was used to detect cardiac function in mice,HE staining and Masson staining were used to observe pathological and morphological changes of the heart,Whole-mount immunofluorescent staining was used to detect the expression of LYVE-1 and Podoplanin in epicardial lymphatic vessels,Western blot was used to detect the expression of VEGFR-3 protein,and qPCR was used to detect the expression of VEGFR-3 mRNA.Results DCM mice induced by DOX showed significant cardiac function decline from the third week(DOX:15 mg·kg-1,P<0.05),and significant ventricular remodeling at the fifth week(DOX:15 mg·kg-1,P<0.01);The lymphatic vessel area of the mouse heart decreased significantly from the fourth week(DOX:20 mg·kg-1,P<0.0001),and the expression of VEGFR-3 decreased significantly from the third week(DOX:15 mg·kg-1,P<0.01).Conclusion KXD can improve ventricular remodeling and cardiac function in DOX-induced DCM mice,promote cardiac lymphangiogenesis,and upregulate the expression of VEGFR-3 at protein and mRNA levels,with a better effect than captopril.DOX-induced cardiac lymphangiogenesis in DCM mice leads to severe myocardial fibrosis and weakened cardiac function,which gradually worsens with the accumulation of modeling time and dose.KXD can promote cardiac lymphangiogenesis and improve cardiac function in DOX-induced DCM mice.The mechanism may be related to the up-regulation of VEGFR-3 expression.

【Objective】 To analyze the polymorphisms of GYPA and GYPB mRNA spliceosomes associated with MNS blood group, and to explore the mechanism of subcellular localization of GPA and GPB protein isomerism encoded by various spliceosomes as well as the expression of MNS blood group antigen. 【Methods】 Ten blood samples of voluntary blood donors were randomly selected. The total mRNA of peripheral blood was extracted and reversed into cDNA. Nested PCR was used to amplify reading open frame of GYPA and GYPB gene, and sequencing was performed by Sanger. The base sequence obtained was compared with GYPA(NCBI: NM_002099) and GYPB(NCBI: Nm_002100.5). After the wild type and various splicing isomer of the open reading frame of GYPA and GYPB had been obtained, they were fused with the encoding gene of green fluorescent protein (GFP) by fusion PCR technology, then cloned and transfected into HEK293 cells for over expression. The subcellular localization of GPA-GFP and GPB-GFP fused fluorescent proteins was monitored by focusing laser scanning microscope. 【Results】 Exon-1 and Exon-2 were missing in GYPA mRNA of the 2 samples, and 2~26 amino acids were missing in the predicted GPA isomer, and the full length sequence of GYPB mRNA was complete. GYPA mRNA was intact in 6 samples, exon-2 was missing in GYPB mRNA, 13~45 amino acids were missing in the predicted GPB protein isomer, and other exon sequences were intact. One sample had intact GYPA mRNA, and 364~385 bases in exon-5 of GYPB mRNA were replaced by AG, indicating truncation of amino acid signal peptide. The GYP mRNA sequences of other samples were complete. The fluorescence signal of GP-GFP fusion protein showed that all GPA and GPB glycoprotein isomers, cloned according to various RNA splicing, could demonstrate the orientation distribution on the cell membrane surface, while some alternative splicing leaded to different degrees of protein dispersion in the cell, and affected the distribution speed and proportion of protein on the cell surface, which might be one of the reasons for the strength variation of MNS antigen. 【Conclusion】 The GYP mRNA spliceosome is obviously polymorphic, but the partial deletion of GYP mRNA fragment does not affect the localization and distribution of the protein isomers encoded by GYP mRNA on the cell surface, which can ensure the expression of MNS antigen characteristics.

Objective This study aimed to establish a pre-metastatic niche mouse model utilizing luciferase-labeled Lewis (Luc-Lewis) lung cancer cells and to assess the efficacy of this model employing both qualitative and quantitative methods. Methods C57BL/6 mice were categorized into two groups: a normal control group and a model group, each containing 15 individual mice. The pre-metastatic niche model was established via tail vein injection of Luc-Lewis lung cancer cells. Body mass were measured daily for all groups. Tumor fluorescence signals within the mice were detected using a high-throughput enzyme marker instrument. Lung tissue specimens were harvested to evaluate metastatic progression. HE staining was used to assess histopathological changes. Real-time quantitative PCR and Western blot analysis were used to detect the mRNA and protein expression of lysyl oxidase (LOX), matrix metalloproteinase 9 (MMP9), versican (VCAN), and fibronectin (FN), which are the specific markers for the formation of the microenvironment of lung tissues before metastasis. Results Significant declines in body mass and observable lethargy were noted in the model group when compared to the control group. Distinct fluorescence signals were observed in the lung tissue of the model group, demonstrating a positive correlation with the duration of model establishment. By day 14, elevated mRNA and protein expression levels of LOX, MMP9, VCAN, and FN were significantly evident. In addition, histopathological evaluations revealed augmented interstitial thickness, alveolar atrophy and significant inflammatory cell infiltration within the lung tissues of the model group. By the 21st day, metastatic lesions manifested in the lung tissues of the model group, suggesting an approximate pre-metastatic niche maturation timeline of 14 days. Conclusion A pre-metastatic niche mouse model for Lewis lung cancer is successfully established.
Mice ; Animals ; Lung Neoplasms/pathology* ; Matrix Metalloproteinase 9 ; Mice, Inbred C57BL ; Carcinoma, Lewis Lung ; Disease Models, Animal ; RNA, Messenger ; Tumor Microenvironment

Although somatic cells can be reprogrammed to pluripotent stem cells (PSCs) with pure chemicals, authentic pluripotency of chemically induced pluripotent stem cells (CiPSCs) has never been achieved through tetraploid complementation assay. Spontaneous reprogramming of spermatogonial stem cells (SSCs) was another non-transgenic way to obtain PSCs, but this process lacks mechanistic explanation. Here, we reconstructed the trajectory of mouse SSC reprogramming and developed a five-chemical combination, boosting the reprogramming efficiency by nearly 80- to 100-folds. More importantly, chemical induced germline-derived PSCs (5C-gPSCs), but not gPSCs and chemical induced pluripotent stem cells, had authentic pluripotency, as determined by tetraploid complementation. Mechanistically, SSCs traversed through an inverted pathway of in vivo germ cell development, exhibiting the expression signatures and DNA methylation dynamics from spermatogonia to primordial germ cells and further to epiblasts. Besides, SSC-specific imprinting control regions switched from biallelic methylated states to monoallelic methylated states by imprinting demethylation and then re-methylation on one of the two alleles in 5C-gPSCs, which was apparently distinct with the imprinting reprogramming in vivo as DNA methylation simultaneously occurred on both alleles. Our work sheds light on the unique regulatory network underpinning SSC reprogramming, providing insights to understand generic mechanisms for cell-fate decision and epigenetic-related disorders in regenerative medicine.
Male ; Mice ; Animals ; Cellular Reprogramming/genetics* ; Tetraploidy ; Pluripotent Stem Cells/metabolism* ; Induced Pluripotent Stem Cells/metabolism* ; DNA Methylation ; Spermatogonia/metabolism* ; Germ Cells/metabolism*