Background and purpose:In 2016 the National Cancer Institute(NCI)decided stopping to use NCI-60 cell lines for drug screening,suggesting that tumor cell lines were losing their value as a tool for drug discovery and basic research.The reason for NCI-60 cells'retirement'was that the preclinical studies based on traditional cellular and animal models did not obtain the corresponding expected efficacy in clinical trials.Since the major cancer behaviors,such as proliferation and metastasis,are fundamentally altered with long-term culture,the tumor cell lines are not representative of the characteristics of cancer in patients.Currently,scientists hope to create a new cancer model that are derived from fresh patient samples and tagged with details about their clinical past.Our purpose was to create patient-derived breast cancer primary cell lines as new cancer model for drug screening and basic research.Methods:Breast cancer tissues were collected in the Department of Breast Surgery,Affiliated Hospital of Guizhou Medical University.The collection of tumor tissue samples was approved by the Ethics Committee of the Affiliated Hospital of Guizhou Medical University(approval number:2022 ethics No.313),and the collection and use of tumor tissues complied with the Declaration of Helsinki.The primary breast cancer cell lines were isolated from the patient's breast cancer tissues and cultured in BCMI medium.After the cells proliferated,the media were replaced with DEME medium.Cell line STR genotyping was done to determine cell-specific genetic markers and identification.Clone formation assay and transplantation assay were done to analyze the ability of breast cancer primary cell lines to form tumors.Results:We created 6 primary breast cancer cell lines.The 6 primary breast cancer cell lines from the patients were tagged with the definitively clinicopathological features,clinical diagnosis,therapeutic regimens,clinical effectiveness and prognostic outcomes.The STR genotyping assays identified the genetic markers and determined the identities of the 6 primary breast cancer cell lines.Clone formation assays and transplantation assay showed that the proliferative capacities of the patient-derived primary breast cancer cell lines were significantly greater compared with the conventional breast cancer cell lines.Conclusion:We created a panel of 6 patient-derived primary breast cancer cell lines as new cancer model for drug screening and basic research in breast cancer.

Objective To evaluate the clinical efficacy and hemorheological effects of traditional Chinese medicine injection combined with conventional Western medicine in the treatment of stable angina pectoris in coronarrt heartdisease using a mesh meta-analysis system.Methods Computer searches were conducted on China National Knowledge Infrastructure(CNKI),WanFang Data Knowledge Service Platform(WanFang Data),VIP,SinoMed,PubMed,Cochrane Library,Web of Science,and other clinical randomized controlled trials(RCTs)related to the combination of traditional Chinese medicine injection and conventional Western medicine for the treatment of stable angina pectoris in coronary heart disease.The search was conducted until April 2023.The Cochrane bias risk assessment tool was used to evaluate the quality of the included studies,and Stata13.0 software was used for meta-analysis.Results 57 RCTs were ultimately included,involving 5 types of traditional Chinese medicine injections.The results of the network meta-analysis showed that:① in terms of clinical total effective rate,the efficacy of traditional Chinese medicine injection was ranked in the following order:conventional Western medicine treatment combined with Shenmai injection>Shenxiong glucose injection>Shenfu injection>Ciwujia injection>Huangqi injection>conventional Western medicine treatment;② In terms of the effective rate of angina symptoms,the order of efficacy of traditional Chinese medicine injection is conventional Western medicine treatment combined with Shenxiong glucose injection>Ciwujia injection+conventional therapy>Huangqi injection>Shenfu injection>Shenmai injection>conventional Western medicine treatment;③ In terms of the effective rate of electrocardiogram,the order of efficacy of traditional Chinese medicine injection is conventional Western medicine treatment combined with Shenmai injection>Shenxiong glucose injection>Huangqi injection>Ciwujia injection>Shenfu injection>conventional Western medicine treatment;④ In terms of improving the high shear viscosity of whole blood,the efficacy of traditional Chinese medicine injection is ranked in the following order:conventional Western medicine treatment combined with Shenmai injection>Shenxiong glucose injection>Shenfu injection>Huangqi injection>Ciwujia injection>conventional Western medicine treatment;⑤ In terms of improving whole blood low shear viscosity,the efficacy of traditional Chinese medicine injections is ranked in the following order:conventional Western medicine treatment combined with Shenfu injection>Shenmai injection>Shenxiong glucose injection>Ciwujia injection>Huangqi injection>conventional Western medicine treatment;⑥ In terms of improving plasma viscosity,the order of efficacy of traditional Chinese medicine injection is conventional Western medicine treatment combined with Huangqi injection>Shenfu injection>Shenmai injection>Shenxiong glucose injection>Ciwujia injection>conventional Western medicine treatment;⑦ In terms of improving fibrinogen,the efficacy ranking of traditional Chinese medicine injection is conventional Western medicine treatment combined with Shenfu injection>Shenxiong glucose injection>Shenmai injection>Ciwujia injection>conventional Western medicine treatment.Conclusion The combination of traditional Chinese medicine injection and conventional Western medicine treatment has a significant effect on stable angina pectoris in coronary heart disease.Among them,Shenmai Injection has the best therapeutic effect in terms of clinical total effective rate,electrocardiogram,and improvement of whole blood high shear viscosity;In terms of improving angina symptoms,Shenxiong Glucose Injection has the best effect;In terms of improving whole blood low shear viscosity and fibrinogen,Shenfu Injection has the best effect;In terms of improving plasma viscosity,Shenfu Injection has the best effect,but currently it is limited by the number and quality of studies included,and the above conclusions still need to be verified by more high-quality studies.

Objective To evaluate the analgesic effects of herbal medicine extraction on bone cancer pain of rat models. Methods Rat models of cancer-induced bone pain was established by using the MRMT-1 cell line injected into the tibia. Changes of behavioral signs indicative of pain including 50% paw withdrawal threshold (von Frey tactile sensitivity test)and thermal withdrawal latency were observed. The cellular reorganization of dorsal root ganglion (DRG) was measured by histological analysis. Results In the behavioural tests, herbal medicine treatment attenuated mechanical allodynia and thermal hyperalgesia. Histological examination showed that herbal medicine inhibited DRG neuronal nuclear and somatic size reduction with nucleolar segregation. Conclusion The herbal medicine extraction was an anti-nociceptive agent in rat models of bone cancer pain.

Objective To evaluate therapeutic effects of Bushen-Zhuanggu medicine extraction on bone destruction in rats bone cancer pain.Methods Rat models of cancer-induced bone pain were established by inoculating MRMT-1 cells into tibia of rats.Behavioral signs indicative of pain including 50%paw withdrawal threshold(von Frey tactile sensitivity test)and thermal withdrawal latency were observed.Pathomorphological changes of tibia were monitored with HE staining.Results In the behavioral tests,herbal medicine treatment attenuated mechanical allodynia and thermal hyperalgesia.Histological examination showed that this treatment inhibited tumor proliferation and preserved the cortical and trabccular bone structure.Conclusion Bushen-Zhuanggu medicine extraction is an anti-nociceptive and bone-preserving agent in rats of bone cancer pain.

Objective To establish the methods for quantitating the circulating tumor DNA with PicoGreen fluorescent nucleic acid stain and investigate the role of the quantitative analysis in diagnosis of breast cancer. Methods Circulating tumor DNA was isolated from serum with QIAmp blood kit and was quantitated by spectrofluorometry with PicoGreen fluorescent stain. Receiver operator characteristic ( ROC) curve and area under the curve were used to estimate the role of DNA quantification in diagnosis of breast cancer. Results The efficiency of QIAamp blood kit isolating DNA from serum was 37. 8%-46. 2%, average 43. 4%. Circulating tumor DNA concentration as low as 1 ng could be detected by PicoGreen spectrofluorometry. and the detected range was 1-500 ng/0. 2 ml. The median concentration of serum DNA in breast cancer group was (169. 70+ 124. 10) μg/L, and that of healthy control and breast benign group were (54. 30±36. 84) μg/L and (51. 70±29. 04) μg/L, respectively (P<0. 01). The area under the ROC curve was 0. 899 (95% CI: 0.848-0.951), and the sensitivity was 78. 2%, the specificity was 90% by using the cutoff value of 96. 0 ng/ml. Conclusions The concentration of circulating tumor DNA can be efficiently quantitated by PicoGreen spectrofluorometry, which indicates the potential of clinical applicability in breast cancer diagnosis.

With Jingkang No.5 (PiA), calli of the PiA induced for 10-15 days were transferred into amino acid liquid culture medium, to establish excellent rice suspension cell lines successfully in a relative short time. The growth characteristics and differentiation conditions of suspension cells were measured at different phases. Results revealed that the optimal subculture time was 7-10 days, and the cells cultured for 30-120 days had the best differentiation ability (57.1%) and regeneration ability (20%). This study is promising in further using the suspension cell for genetic transformation and protoplasm isolation.
Cell Culture Techniques ; Cell Differentiation ; physiology ; Cell Line ; Culture Media ; Oryza ; cytology

Objective To study and evaluate the effects of Kanglaite Injection (KLT) in rat model of cancer pain. Methods According to Medhurst' method, a rat model of cancer-induced bone pain wasestablished by intra-tibial injection of 3?103 MRMT-1 rat mammary gland carcinomal cells in Sprague-Dawley rats. KLT group were treated with 10 mL/kg of KLT, ip, twice daily from day 8 to day 17 after injection of MRMT-1 cells. In celecoxib group, 30 mg/kg, ig, twice daily for 10 days. NS group and sham group were given with 10 mL/kg of NS, ip, twice daily for 10 days. On day 17, rats in morphine group received a single administration of 3 mg/kg morphine. Pain-related behaviors including allodynia (the hind paw withdrawal response to 2 g von Frey filament stimulation) and the difference of weight bearing between hind paws were measured in all 5 groups on day 17. In addition, the left hind limbs were removed and X-rays were obtained for assessing of tumor-induced bone destruction. Results On day 17 after injection of MRMT-1 cells or NS, the paw withdrawal response frequency to 2 g von Frey filament stimulation on the left paw in KLT group, celecoxib group, morphine group and NS group were 53.3%?20.7%, 46.7%?20.7%, 13.3%?16.3% and 90% ?16.7%, respectively. The paw withdrawal response frequency in KLT group was significantly decreased compared with that in NS group (P

Objective To investigate the effect of Huangqikeli on immune function of mice. Methods 30 mice were randomly divided into 3 groups. The mice in A and B group were administrated with distilled water,C group were administrated with Huangqikeli. On the tenth day,all groups except A group were injected with cyclophosphamide to induce the model. The effects on the phagocytic function of Huangqikeli to macrophage in abdominal cavity of mice,delayed hypersensitivity induced by DNCB,and production of hemolysin antibody in mice that CRBC sensitized were observed. Result Compared with immuno-suppressive models given distilled water (B group),phagocytic function of macrophage in abdominal cavity of mice and delay hypersensitivity leaded by DNCB were enhanced (P

Objective To investigate the influence of Yudan Rongxin Keli on immunoregulation in mice. Methods The mice were randomly divided into the normal group, cyclophosphamide (Cy) model group, Yudan Rongxin Pill group and Yudan Rongxin Keli group. The effects of Yudan Rongxin Keli on the phagocytosis of macrophage in mouse abdomen, delayed-type hypersensitivity (DTH) induced by dinitrochlorobenzene (NDCB), and generation of hemolysin antibody induced by chicken red blood cell (CRBC) were observed respectively. Result Compare with the Cy model group, the phagocytosis of macrophage in Yudan Rongxin Keli group was promoted (P

Objective:To evaluate the anti-metastatic and bone preserving therapeutic effects of herbal medicine extraction on rat model of bone metastatic carcinoma.Methods:A rat model of cancer-induced bone pain using the MRMT-1 cell line injected into the tibia was established to investigate the efficacy of the herbal medicine extraction,on osteoclast activity and bone mineral density.The development of the bone tumor and structural damage to the bone was monitored by radiological analysis.Specimens of the tibial bone were processed for tartrate-resistant acid phosphatase(TRAP)stain to observe the bone pathological changes and count TRAP stained osteoclasts.OPG and RANKL expression was evaluated by immunohistological methord.Results:Histological and radiological examination showed that the herbal medicine extraction significantly inhibited tumor proliferation and preserved the cortical and trabecular bone structure.In addition,a dramatic reduction of tartrate resistant acid phosphatase-positive polykaryocytes(osteoclasts)and increase of OPG expression were observed.Conclusions:The herbal medicine extraction was an anti-metastatic and bone preserving therapeutic effects in a rat model of metastatic cancer pain.