Altered Splicing in Stable Cell Strains Expressing Mini-hF9 Gene with Nonsense Mutation / 中国实验血液学杂志

OBJECTIVE@#To investigate the molecular mechanism in stable cell strains expressing Mini-hF9 gene with nonsense mutation.@*METHODS@#Mini-hF9 gene and its nonsense mutants were transfected into HeLa cells independently, and stable cell strains were obtained after G418 resistance screening and monoclonal transformation. The altered splicing and protein expression of mRNA in Mini-hF9 gene in stable cell strains were detected by using RT-PCR and Western blot.@*RESULTS@#The wild type and nonsense mutated human coagulation factor IX stable cell strains were constructed successfully, which were named HeLa-F9-WT, HeLa-F9-M1 and HeLa-F9-M2. Only normal splicing Norm was detected in the wild-type cell strain HeLa-F9-WT; Norm and Alt-S1 splicing were detected in HeLa-F9-M1; while Norm, Alt-S1 and Alt-S2 splicing were detected in HeLa-F9-M2.@*CONCLUSION@#The nonsense associated altered splicing (NAS) pathway, which generated alternately spliced transcripts, might be triggered in coagulation factor IX gene with nonsense mutation.

Clinical value of contrast-enhanced ultrasound-guided biopsy in early diagnosis of pleural tuberculoma / 第二军医大学学报

Objective To explore the clinical value of contrast-enhanced ultrasound-guided biopsy in the early diagnosis of pleural tuberculoma. Methods A total of 87 patients were recruited, who were admitted to Shanghai Pulmonary Hospital affiliated to Tongji University from Jan. 2017 to Jan. 2018 and were diagnosed to have pleural tuberculoma based on clinical indication and imaging. All patients underwent contrast-enhanced ultrasonography and ultrasound-guided biopsy, and then the samples were sent for pathological examination, acid-fast bacilli culture and polymerase chain reaction for Mycobacterium tuberculosis. The clinical data of the patients were collected and the results of contrast-enhanced ultrasound, final diagnostic positive rate and adverse reactions were statistically analyzed. Results The success rate of contrast-enhanced ultrasound-guided biopsy of 87 patients was 100%, among which 70 cases were diagnosed with tuberculosis, 7 with maligmant tumors, 4 with other infection diseases, and 6 cases obtained negative results (confirmed by diagnostic anti-tuberculosis and thoracoscopic biopsy, 5 with tuberculosis and 1 with pleural mesothelioma). Seventy-five of 87 cases were eventually diagnosed with tuberculosis, of which 70 (93.33%) were diagnosed by contrast-enhanced ultrasound-guided biopsy. The intensity of contrast-enhanced ultrasound was an important differential factor for pleural tuberculoma and malignant tumor; low enhancement mainly existed in the pleural tuberculoma (66/75), and high enhancement mainly existed in the malignant tumor (4/7). Hemorrhage and/or hemoptysis occurred in 5 cases during the puncture, pleural reaction occurred in 3 cases, and all of them was relieved after symptomatic treatment. Conclusion Contrast-enhanced ultrasound-guided biopsy has a high value for the early diagnosis of pleural tuberculoma, with high positive rate, small trauma and good reproducibility, and is worthy of clinical promotion.

Killing effect of Huaier combined with DC-CIK on nude mice bearing colon cancer HT29 stem cells / 中国中药杂志

To compare the therapeutic effects of different treatment methods on the nude mice bearing colon cancer HT29 cells. BalB/C nude mice colon cancer stem cell models were established and randomly divided into the following four groups, with 8 nude mice in each group: blank control group, DC-CIK group, Huaier group, and Huaier combined with DC-CIK group (combined treatment group). The mice in DC-CIK group and combined treatment group received 1×10⁶ DC-CIK cells treatment by tail vein injectionafter the tumor stem cells were inoculated for 4 days,2 times a week for three weeks. The mice in Huaier group and combined treatment group received intragastric administration at the dose of 20 g/60 kg body weight, 0.2 mL/time, once a day for a total of three weeks. The mice in control group received equal volume of normal saline. Tumor size and body weight of nude mice were measured every 2 days during treatment for three weeks in each group. After the treatment, the nude mice were sacrificed to measure the tumor weight and the tumor inhibition rate was calculated. The RT-PCR method was used to detect the expression levels of the key genes in the signal pathway. After the end of the treatment, the quality of the tumor in the Huaier group, DC-CIK group and combined treatment group was significantly lower than that in the control group; the quality in combined treatment group was significantly lower than that in Huaier group and DC-CIK group.Among them, the tumor inhibition rate reached 46.77% in the combined treatment group. In respect of changes in expression levels of key genes in the signaling pathway, the mRNA expression levels of key genes PI3KR1 and Akt in PI3K/Akt pathway, key genes Wnt1 and CTTNB1 in Wnt/-catenin pathway, and key genes Notch1, Notch2, Notch3 in Notch pathway in the combined treatment group were lower than those in DC-CIK group and Huaier group. The Huaier combined with DC-CIK group showed best therapeutic effect among different treatment methods for HT29 stemcell colon tumors in nude mice, providing a new idea for clinical treatment of colon cancer.

The effect of Chinese yam extract combined with tegafur on the cytotoxicity of HT-29 cell line in vivo and in vitro / 天津医药

Objective To study the inhibitory effects of Chinese yam combined with tegafu on colon cancer HT-29 cells in vivo and in vitro.Methods The changes of cell morphology were observed by the method of adopting 2×2 factorial designs,after HT-29 colon cancer cells were treated with physiological saline containing dimethyl sulfoxide(blank control), tegafur(36 mg/L)and Chinese yam(125 mg/L).MTT method was used to detect the proliferation of tumor cells,and flow cytometry was used to detect the expression of tumor stem cells(CD133+cells).The nude mouse model of colon cancer HT-29 cells was established. The treatment was given in the above method, and the tumor inhibition rate was calculated. The VEGF positive rate of the tumor was detected by immunohistochemistry. Results The inhibited proliferation of colon cancer HT-29 cells was significantly higher in the synergistic group than that in Chinese yam group and tegafur group,and the inhibitory rates were higher in the three treatment groups than that of blank control group.The proportion of CD133+cells was significantly lower in HT-29 cells in the synergistic group compared with that of the Chinese yam group and the tegafur group,and which was lower in these three groups than that of the blank control group.After the treatment in three treatment groups,the tumor quality and VEGF positive rate were significantly lower than those of the blank control group,and which was lower in the tegafur+Chinese yam group than that of Chinese yam group and the tegafur group. Conclusion Chinese yam combined with tegafu can inhibit the proliferation of colon cancer HT-29 cells and their stem cells.

In vivo antitumor effect of Chinese yam extract combined with DC-CIK on nude mice bearing colon cancer HT29 stem cells / 中草药

Objective: To compare the therapeutic effects of different treatments on the nude mice bearing colon cancer HT29 cells. Methods: BalB/C nude mice colon cancer stem cell model was established and the mice were randomly divided into following four groups: blank control group, DC-CIK group, yam group, and yam combined with DC-CIK group (combined treatment group), each group of 10 nude mice. In DC-CIK group and combined treatment group, after tumor stem cells were inoculated in nude mice for 4 d, 1 × 106 DC-CIK cells were used through the tail vein injection for treatment, twice a week for three weeks. The mice in Chinese yam group and combined treatment group were ig administered with 125 mg/kg Chinese yam extract, once daily for three weeks, and the mice in control group were treated with equal volume of saline instead. Tumor size and body weight of nude mice were measured every 2 d during treatment for three weeks in each group. After the treatment, the nude mice were sacrificed to take out the tumor weight and the tumor inhibitory rate was calculated. The RT-PCR method was used to detect the expression levels of the key genes in signal pathway. Results: After treatment, the quality of tumor in yam group, DC-CIK group and combined treatment group was significantly lower than that in control group. Among them, the inhibitory rate was 51.26% in combined treatment group. In the changes of expression levels of key genes in signaling pathway, the key genes of PI3KR1 in PI3K/Akt pathway, the key gene of Wnt1 in Wnt/beta-catenin pathway, and the mRNA expression of key gene Notch1 in Notch pathway in combined treatment group were decreased compared with DC-CIK group and yam group. Compared with the DC-CIK group, the mRNA expression changes were not statistically significant. Conclusion: The effect of Chinese yam combined with DC-CIK group is the best on the treatment of HT29 cell stem cell tumor of colon cancer in nude mice. It provides a new idea for clinical treatment of colon cancer.

Lymphoma of nasal cavity and nasopharynx: a report of 319 consultation cases / 临床与实验病理学杂志

Purpose To investigate the clinical features,pathological type and composition of nasal cavity and nasopharyngeal lymphoma.Methods The clinical data,histopathologic features and immunotypes of 319 consulted cases of nasal cavity and nasopharyngeal lymphoma from department of pathology,Beijing Friendship Hospital,Capital Medical University were retrospectively analyzed.According to the new WHO (2008) classification,all data of these cases were reappraised.Results Among these 319 cases,3 cases (0.9％) were diagnosed as classical Hodgkins lymphoma (CHL).Other 316 cases (99.1％) were diagnosed as non-Hodgkins lymphoma (NHL),56.0％ (177 cases) of them were T and NK-cell lymphoma and 44.0％ (139 cases) were B-cell lymphoma.The commonest subtypes were extra-nodal NK/T-cell lymphoma (NK/TCL) in 160 cases (50.6％),diffuse large B-cell lymphoma in 64 cases (DLBCL 20.3％).Among these 319 cases,106 cases (33.2％) over 60 years old were diagnosed as NHL.The most common subtypes were DLBCL in 40 cases (37.7％),NK/TCL in 36 cases (34.0％).There was no statistic difference in the incidence rate between the two types.Other 210 cases of NHL 0.9％ below 60 age including NK/TCL 124 cases (59.0％)and DLBCL 24 cases (11.4％).The diference is statistically significant.Conclusion Among those 319 nasopharyngeal lymphoma cases,3 cases are considered as CHL.NK/TCL is the commonest subtype of other 316 cases.DLBCL and NK/TCL are the most common subtypes in 106 cases over 60 years old.There was no statistic difference in the incidence rate between the two types.