[Preparation and in vitro evaluation of FDM 3D printed theophylline tablets with personalized dosage].

OBJECTIVE: To explore the feasibility of preparing different doses of tablets for personalized treatment by fused deposition modeling (FDM) 3D printing technology, and to evaluate the in vitro quality of the FDM 3D printed tablets. METHODS: Three different sizes of hollow tablets were prepared by fused deposition modeling 3D printing technology with polyvinyl alcohol (PVA) filaments. Theophylline was chosen as the model drug. In the study, 20 mg, 50 mg and 100 mg of theophylline was filled into the cavity of the tablets, respectively. The microscopic morphology of the tablets was observed by scanning electron microscopy (SEM). The weight variation of the tablets was investigated by weighing method. The hardness of the tablets was measured by tablet hardness tester. The contents of the drugs in the tablets were determined by ultraviolet and visible spectrophotometry (UV-Vis), and the dissolution apparatus was used to assay the in vitro drug release of the tablets. RESULTS: The prepared FDM 3D printed tablets were all in good shape without printing defects. And there was no leakage phenomenon. The diameter and thickness of the tablets were consistent with the design. The layers were tightly connected, and the fine structure of the formulation could be clearly observed without printing defects by scanning electron microscopy. The average weight of the three sizes of tablets was (150.5±2.3) mg, (293.6±2.6) mg and (456.2±5.6) mg, respectively. The weight variation of the three sizes of tablets were boss less than 5%, which met the requirements; The hardness of the tablets all exceeded 200 N; The contents of theophylline in the three tablets were 98.0%, 97.2% and 97.9% of the dosage (20 mg, 50 mg and 100 mg), and the relative standard deviation (RSD) was 1.06%, 1.15% and 0.63% respectively; The time for 80% drug released from the three dosage of tablets was within 30 min. CONCLUSION: Three different dosages of theophylline tablets were successfully prepared by FDM 3D printing technology in this study. The exploration may bring beneficial for the preparation of personalized dose preparations. We expect that with the development of 3D printing technology, FDM 3D printed personalized tablets can be used in the clinic as soon as possible to provide personalized treatment for patients.

[Preparation and in vitro evaluation of fused deposition modeling 3D printed compound tablets of captopril and hydrochlorothiazide].

OBJECTIVE: To explore the feasibility of preparing compound tablets for the treatment of hypertension by fused deposition modeling (FDM) 3D printing technology and to evaluate the quality of the printed compound tablets in vitro. METHODS: Polyvinyl alcohol (PVA) filaments were used as the exci-pient to prepare the shell of tablet. The ellipse-shaped tablets (the length of major axes of ellipse was 20 mm, the length of the minor axes of ellipse was 10 mm, the height of tablet was 5 mm) with two separate compartments were designed and printed using FDM 3D printer. The height of layer was 0.2 mm, and the thickness of roof or floor was 0.6 mm. The thickness of shell was 1.2 mm, and the thickness of the partition wall between the two compartments was 0.6 mm. Two cardiovascular drugs, captopril (CTP) and hydrochlorothiazide (HCT), were selected as model drugs for the printed compound tablet and filled in the two compartments of the tablet, respectively. The microscopic morphology of the tablets was observed by scanning electron microscopy (SEM). The weight variation of the tablets was investigated by electronic scale. The hardness of the tablets was measured by a single-column mechanical test system. The contents of the drugs in the tablets were determined by high performance liquid chromatography (HPLC), and the dissolution apparatus was used to measure the in vitro drug release of the tablets. RESULTS: The prepared FDM 3D printed compound tablets were all in good shape without printing defects. The average weight of the tablets was (644.3±6.55) mg. The content of CTP and HCT was separately (52.3±0.26) mg and (49.6±0.74) mg. A delayed in vitro release profile was observed for CTP and HCT, and the delayed release time for CTP and HCT in vitro was 20 min and 40 min, respectively. The time for 70% of CTP and HCT released was separately 30 min and 60 min. CONCLUSION: CTP and HCT compound tablets were successfully prepared by FDM 3D printing technology, and the printed tablets were of good qualities.

MiR-1266 suppresses the growth and metastasis of prostate cancer via targeting PRMT5.

The article "MiR-1266 suppresses the growth and metastasis of prostate cancer via targeting PRMT5, by C.-M. Sun, G.-M. Zhang, H.-N. Qian, S.-J. Cheng, M. Wang, M. Liu, D. Li, published in Eur Rev Med Pharmacol Sci 2019; 23 (15): 6436-6444-PMID: 31378882" has been withdrawn from the authors due to some inaccuracies (some data cannot be repeated by our further research). The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/18525.

Circular RNA hsa_circ_0017247 acts as an oncogene in bladder cancer by inducing Wnt/ß-catenin signaling pathway.

The article "Circular RNA hsa_circ_0017247 acts as an oncogene in bladder cancer by inducing Wnt/ß-catenin signaling pathway, by C.-T. Han, Q.-Y. Bao, S.-J. Cheng, M. Liu, H.-N. Qian, D. Li, published in Eur Rev Med Pharmacol Sci 2020; 24 (3): 1081-1087-DOI: 10.26355/eurrev_202002_20158-PMID: 32096177" has been withdrawn from the authors since they decided to perform further experiments. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/20158.

[Preparation and in vitro evaluation of fused deposition modeling 3D printed verapa-mil hydrochloride gastric floating formulations].

OBJECTIVE: To explore the feasibility of preparing gastric floating formulations by fused de-position modeling (FDM) 3D printing technology, to evaluate the in vitro properties of the prepared FDM 3D printed gastric floating formulations, and to compare the influence of different external shapes of the formulation with their in vitro properties. METHODS: Verapamil hydrochloride and polyvinyl alcohol (PVA) were used as the model drug and the excipient, respectively. The capsule-shaped and hemisphere-shaped gastric floating formulations were then prepared by FDM 3D printing. The infill percentages were 15%, the layer heights were 0.2 mm, and the roof or floor thicknesses were 0.8 mm for both the 3D printed formulations, while the number of shells was 3 and 4 for capsule-shaped and hemisphere-shaped formulation, respectively. Scanning electron microscopy (SEM) was used to observe the morpho-logy of the surface and cross section of the formulations. Gravimetric method was adopted to measure the weights of the formulations. Texture analyzer was employed to evaluate the hardness of the formulations. High performance liquid chromatography method was used to determine the drug contents of the formulations. The in vitro floating and drug release behavior of the formulations were also characterized. RESULTS: SEM showed that the appearance of the FDM 3D printed gastric floating formulations were both intact and free from defects with the filling structure which was consistent with the design. The weight variations of the two formulations were relatively low, indicating a high reproducibility of the 3D printing fabrication. Above 800.0 N of hardness was obtained in two mutually perpendicular directions for the two formulations. The drug contents of the two formulations approached to 100%, showing no drug loss during the 3D printing process. The two formulations floated in vitro without any lag time, and the in vitro floating time of the capsule-shaped and hemisphere-shaped formulation were (3.97±0.41) h and (4.48±0.21) h, respectively. The in vitro release of the two formulations was significantly slower than that of the commercially available immediate-release tablets. CONCLUSION: The capsule-shaped and hemisphere-shaped verapamil hydrochloride gastric floating formulations were prepared by FDM 3D printing technology successfully. Only the floating time was found to be influenced by the external shape of the 3D printed formulations in this study.

Circular RNA hsa_circ_0017247 acts as an oncogene in bladder cancer by inducing Wnt/ß-catenin signaling pathway.

OBJECTIVE: Bladder cancer (BLCA) is the most common genitourinary malignancy in the world. Recent studies have revealed that circular RNAs (circRNAs) are dysregulated in malignant tumors and participate in carcinogenesis. The purpose of our work is to uncover how hsa_circ_0017247 functions in BLCA. PATIENTS AND METHODS: In this research, Real Time-quantitative Polymerase Chain Reaction (RT-qPCR) was conducted to monitor hsa_circ_0017247 expression in BLCA samples. Besides, proliferation assay, colony formation assay, and flow cytometry assay were performed in BLCA cells after hsa_circ_0017247 was knocked down. Meanwhile, the Western blot assay was conducted to explore the target signaling pathway of hsa_circ_0017247. Furthermore, tumor formation and metastasis assays were also conducted in vivo. RESULTS: Compared with the adjacent tissues, a significant upregulation in hsa_circ_0017247 expression was observed in BLCA samples. Functional assays showed that the inhibition of cell proliferation was induced via downregulating hsa_circ_0017247 in BLCA in vitro, while the promotion of cell proliferation was induced via downregulating hsa_circ_0017247 in BLCA in vitro. Moreover, the results of further experiments revealed that the targeted proteins in the Wnt/ß-catenin signaling pathway were downregulated via knockdown of hsa_circ_0017247 in BLCA. In addition, tumor formation and metastasis of BLCA were inhibited via knockdown of hsa_circ_0017247 in nude mice. CONCLUSIONS: We discovered a vital regulatory mechanism of hsa_circ_0017247 in BLCA which might serve as a new therapeutic intervention for BLCA patients.

MiR-1266 suppresses the growth and metastasis of prostate cancer via targeting PRMT5.

OBJECTIVE: To elucidate the correlation between microRNA-1266 (miR-1266) and prostate cancer (PCa) progression, and to investigate the possible underlying mechanism. PATIENTS AND METHODS: The expression level of miR-1266 and protein arginine methyltransferase 5 (PRMT5) in PCa tissues and cell lines was first detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). After up-regulating or down-regulating miR-1266 expression in cells, cell proliferation, migration and invasion abilities were detected. Possible target genes of miR-1266 were predicted and validated by bioinformatics analysis and dual-luciferase reporter gene assay, respectively. Finally, abnormal expression of PRMT5 was ascertained after transfection. RESULTS: MiR-1266 was lowly expressed in PCa tissues and cell lines, whereas PRMT5 exhibited the opposite results. Up-regulated expression of miR-1266 significantly inhibited the proliferation, migration and invasion abilities of PC-3 cells. However, the growth and migration of DU145 cells with low miR-1266 expression were significantly accelerated. Meanwhile, the number of invading cells was significantly increased. PRMT5 was verified as a potential target gene of miR-1266. Furthermore, results found that miR-1266 was negatively correlated with PRMT5. In addition, the expression of PRMT5 was remarkably decreased after miR-1266 overexpression, which could be restored after knockdown of miR-1266. CONCLUSIONS: MiR-1266 inhibits the growth and metastasis of PCa by targeting PRMT5. We may provide a potential and prospective therapeutic target for PCa.

The anti-tumor immune responses induced by a fusion protein of ovarian carcinoma anti-idiotypic antibody 6B11ScFv and murine GM-CSF in BALB/c mice.

Ovarian carcinoma anti-idiotypic antibody 6B11 was murine derived; we previously have cloned 6B11 single-chain Fv antibody (6B11ScFv) and constructed the 6B11ScFv/human granulocyte-macrophage colony stimulating factor (GM-CSF) fusion protein (designated as 6B11GM) to enhance the immunogenecity of the single-chain Ab(2). Because of the difference in species specificity between human GM-CSF and murine GM-CSF, there is no immune competent animal model on which the effect and metabolism of 6B11GM as a vaccine could be observed. In this study, 6B11mGM fusion gene was constructed by the fusing murine GM-CSF cDNA gene with 6B11ScFv. The fusion gene was cloned and expressed. The product of this gene is a fusion protein. It could specifically interact with the primary anti-ovarian carcinoma monoclonal antibody (COC166-9) and rat anti-mouse GM-CSF monoclonal antibody, respectively, and stimulate the growth of NFS-60 cells (a murine GM-CSF-dependent cell line). The specific anti-tumor immune response could be induced in BALB/c mice after immunized with anti-idiotypic fusion protein instead of ovarian carcinoma antigen without carrier proteins and adjuvant. Ab(3) could be detected in the sera of immunized mice with 6B11mGM by enzyme-linked immunoadsorbent assay test. Moreover, the fusion protein stimulated proliferation of CD4+ T cell from the spleen of BALB/c mice and proliferation of CD8+ T cell to a lesser degree. Therefore, 6B11mGM probably induces both humoral and cellular immunity against ovarian carcinoma in vivo.

The experimental study of ovarian carcinoma vaccine modified by human B7-1 and IFN-gamma genes.

With the advance of immunology, immunogene therapy is becoming a possible therapeutic alternative to advanced cancer. The aim of tumor immunogene therapy is to enhance the immune response to tumors. Evidence suggests that CD80 (B7-1) and Interferon-gamma (IFN-gamma) play important roles in antitumor immunity induced by T lymphocyte. To study the antitumor immune effects of ovarian carcinoma vaccine modified with human B7-1 and IFN-gamma genes, we constructed the bicistronic retroviral vector pLXSN, encoding human B7-1 and IFN-gamma. In vitro, the primary ovarian carcinoma cells were transduced with the retroviral vector and prepared as a vaccine. Then autologous lymphocytes were irritated with the ovarian carcinoma cells for competition inhibitory cytotoxic testing. The tumor-specific cytotoxic activity was greatly enhanced by the double gene-modified vaccine. In vivo, the tumorigenicity of the double gene-modified ovarian cell line 3AO/B7-1.IFN-gamma was evaluated in a human immune reconstituted SCID mice (hu-PBL-SCID) model. The double-gene modification markedly decreases tumor genecity. Together, the results suggest that combining B7-1 and IFN-gamma could be a useful therapeutic approach in ovarian cancer.

[Cloning of human TRAIL cDNA and its expression in COS-7 cells].

The human TRAIL cDNA was amplified with the total RNA from the human acute promyelocytic leukemia cell line HL-60 by means of RT-PCR, and was cloned into the pGEM-T vector. The DNA sequence analysis showed that it was consistent with the published sequence. Then, the insert of human TRAIL cDNA was subcloned into the mammalian expression vector pcDNA3. The hybrid plasmid pcDNA3-hTRAIL was transformed into COS-7 cells, and transiently expressed in the COS-7 cells. The activity of the expressed product could induce apoptosis in U937 cell line.

[Analysis of complications associated with catheter intraperitoneal chemotherapy of ovarian carcinoma].

115 patients suffering from ovarian carcinoma were admitted from May 1976 through August 1991, and were treated with intra-peritoneal chemotherapy. A total of 191 catheters which with 2mm in diameter and 50 cm in length were inserted with 608 courses of chemotherapy. 29 plastic catheters used by 29 patients were kept for 18.4 days averagely, while 162 silica catheters used by 86 patients were kept 109 days in average. The duration of keeping silica catheters was significantly longer than plastic catheters (P < 0.01). Complications were found in 29 patients, 25.2% of the total: 5 cases of infection (4.3%), 2 of partial intestinal obstruction (1.7%), 4 of painful sensation (3.5%), 12 with inflow obstruction (10.4%) and 6 with falling off of the catheters (5.2%). Complications between the two groups were compared. There was no statistical significance (P < 0.05). When catheter retainment times of the two groups were compared, significant differences were found between plastic and silica catheters (P < 0.001). Our results indicate that both kinds of catheters may be used in intraperitoneal chemotherapy of ovarian cancer patients, and the silica ones seem better.

[Target therapy by monoclonal antibody against ovarian carcinoma conjugated with liposomes and adriamycin].

Conjugates of monoclonal antibody COC166-9 and adriamycin entrapped in liposome (MLA) were prepared in our laboratory. In vitro growth inhibition of SKOV3 ovarian carcinoma cell line was carried out by MLA and controls. Target therapies by MLA, adriamycin, normal mouse IgG instead of MAb, and control were given for 24 nude mice models with subcutaneous human ovarian carcinoma xenografts and 16 ascitic ovarian carcinoma respectively. MLA group showed the best therapeutic effect than all the other groups which gave a helpful clue to clinical use.

[Target therapy by monoclonal antibody against ovarian carcinoma conjugated with liposome and adriamycin].

Conjugates of monoclonal antibody (MAb) and adriamycin entrapped in liposome (MLA) were prepared by COC166-9. The MAb against ovarian serous adenocarcinoma was generated in our laboratory. Target therapies were done in nude mice model with subcutaneous tumor xenografts in 24 and ascitic carcinoma in 16 by MLA. The results demonstrated that MLA group presented the best therapeutic effect than all the other groups, which gave a very helpful clue to clinical target therapy of ovarian carcinoma in the future.

Anti-idiotypic monoclonal antibodies against anti-ovarian carcinoma monoclonal antibody COC166-9. Generation and application.

Anti-idiotypic monoclonal antibody (Mab Ab2) by MAb COC166-9 against ovarian serous papillary adenocarcinoma was prepared. Hybridomas of Ab2 screened by sandwich ELISA and immunocompetitive inhibition tests were procured and named as 6B11 and 1H12. The number of their chromosomes were 93 and 91, and DNA analysis also proved the characteristics of hybridomas. These Ab2s could induce delayed type hypersensitivity (DTH), the cellular immune response. The results of the immune reaction of 6B11 with SKOV3 (ovarian carcinoma cell line) were similar to OC166-9 (Ag), the positive control, while 1H12 was weaker. Anti-anti-idiotypic antibody (Ab3) was also raised by 6B11 and 1H12 respectively. They all showed positive immunohistochemical stainings with ovarian serous adenocarcinoma tissue sections and immunocytochemical stainings with SKOV3 cells as was shown by COC166-9. In the antibody dependent cell mediated cytotoxicity (ADCC) tests, they showed no differences against SKOV3 as compared with COC166-9. We anticipate that 6B11 and 1H12 may be used as vaccines against ovarian carcinoma and may provide a clue for its prevention and treatment.

[The induction of specific immunity to human ovarian carcinoma cells by anti-idiotypic antibodies].

Monoclonal anti-idiotypic antibodies, 6B11 and H12, were employed instead of the tumor antigen to induce a specific cellular immunity to ovarian carcinoma cell line SKOV3. 6B11 and 1H12 carring the internal image of ovarian carcinoma antigen. BALB/c mice were immunized with 6B11 or 1H12 and then the mouse foot pad was attacked by the injection of target cell SKOV3 to stimulate a specific delayed-type hypersensitivity (DTH). The results showed that 6B11 induced a severe foot pad swelling (Mean thickness 0.93mm) as seen in the positive control (P > 0.05) induced by ovarian carcinoma antigen; the 1H12 induced swelling was significantly lower than the positive control (P < 0.05). Both 6B11 and 1H12 expressed the effective responses only to SKOV3, but not to the unrelated target cells. Pathological examinations found that there was marked infiltration of inflammatory cells in foot pad of mice primed by 6B11, while by 1H12, only induced a narrow pathological changes. These observations indicated that there would be some essential differences between 6B11 and 1H12 in the behavior of inducing the specific DTH to SKOK3. And as an idiotypic vaccine, 6B11 would be more powerful against ovarian carcinoma.

[Antigen mimicry of anti-idiotypic antibodies for human ovarian carcinoma antigen: generation and characterization of anti anti-idiotype antibodies].

The antigen mimicry of monoclonal anti-idiotypic antibodies 6B11 and 1H12 was investigated, which carrying the internal image of human ovarian carcinoma antigen. Anti-anti-idiotypic antibodies, obtained from the mice immunized with 6B11 or 1H12, binded to ovarian tumor antigen OC 166-9 specifically by enzyme linked immunosorbent assay (ELISA). Western blot analysis showed that Ab3 and Ab1 recognized the same antigenic protein. Through antibody dependent cell-mediated cytotoxicity (ADCC), Ab3 sera killed the ovarian carcinoma cell line SKOV3 specifically in vitro. These results suggest that 6B11 and 1H12 may substitute for the nominal antigen to stimulate a specific immune response and that they are potential candidates as idiotype vaccines against ovarian carcinomas.

Target therapy of ovarian carcinoma by monoclonal antibodies bearing chemical drugs entrapped in liposomes.

Monoclonal antibody COC166-9 against ovarian carcinoma was conjugated with adriamycin or cisplatinum entraped in liposomes as immunochemical liposomes MLA and MLP respectively. MLA was shown to have the highest effect than adriamycin or other groups on SKOV3 (ovarian cancer cell line) growth inhibition. MLA was also used in target therapy on nude mice bearing human ovarian carcinoma by xenograft of SKOV3 cells. The observations of tumor volumes revealed that this target therapy other than the controls presents a significantly better result which gives a hopeful clue to ovarian carcinoma treatment.

[Human ovarian epithelial adenocarcinoma with peritoneal metastasis and ascites established in murine models].

By means of SKOV3 subcutaneous xenografts in nude mice, both carcinoma cells and cell suspensions were administrated intraperitoneally to establish a model of metastatic ovarian carcinoma with ascites. The intraperitoneal metastasis and cancer cells in the ascitic fluid were all prove to be adenocarcinoma by microscopic and electromicroscopic examinations. The immunohistochemical and immunocytochemical stainings with monoclonal antibody COC166-9 were the same as SKOV3 cells. The median survival of these nude mice was 68.7 days. This may serve as a good experimental model for testing chemotherapy as well as immunotarget therapy. Similar results obtained by xenografts of OSC2. 40 nude mice were included in this study.

Genital carcinoma after radiotherapy for carcinoma of the cervix. Report of two cases.

Two cases of genital carcinoma were found after radiotherapy for carcinoma of the cervix. Vulval carcinoma occurred 30 years after radiation with initial symptoms of itching and whitish changes of the skin at the external genitalia. Adenocarcinoma of the endometrium occurred 9 years after radiation and diffused intra-abdominal metastasis was found surgically. The sites of second malignancies following irradiation for cervical carcinoma and the time interval between them were reviewed. The characteristics of the post-radiation vulval carcinoma and the endometrial carcinoma were discussed.