Objective To investigate vascular endothelial growth factor-c (VEGF-C) and vascular endothelial growth factor receptor-3(VEGFR-3) mRNA expression, microvessels density (MVD) and lymphatic microvessels density (LVD) in human hepatocellular carcinoma and normal liver tissue. Try to illuminate the relationship among VEGF-C,VEGFR-3,MVD,LVD and the clinical pathological features of hepatocellular carcinoma. Methods Liver tissue of 60 cases definitely diagnosed as hepatocellular carcinoma and 20 normal cases were collected. VEGF-C and VEGFR-3 mRNA expression were examined by RT-PCR, MVD and LVD were examined by immunohistochemistry staining. Relationship between these indexes and clinical pathological features of hepatocellular carcinoma was also analysed. Results VEGF-C and VEGFR-3 mRNA expression, MVD and LVD in hepatocellular carcinoma were higher than those in normal liver tissue (P<0.01); In hepatocellular carcinoma tissue, expression of VEGF-C mRNA positively related with VEGFR-3 mRNA, MVD and LVD(P<0.01). VEGF-C and VEGFR-3 expression positively related with portal vein tumor thrombus, intrahepatal metastasis and lymph node metastasis (P<0.01). MVD positively related with portal vein tumor thrombus and intrahepatal metastasis (P<0.01). LVD positively related with lymph node metastasis (P<0.01). Conclusion VEGF-C and VEGFR-3 expression increase in hepatocellular carcinoma tissue. They might play roles in tumor invasion and metastasis by inducing angiogenesis and lymphangiogenesis.

OBJECTIVE:To establish a method for the uncertainty evaluation of determining content of Lamivudine tablets by HPLC. METHODS:HPLC was established to determine the mathematical model of Lamivudine tablets contents,analyze the uncer-tainty sources in detail and evaluate each uncertainty components and combined uncertainty to obtain the expanded uncertainty. RE-SULTS:Uncertainty of Lamivudine tablets contents determined by HPLC was mainly from reference purity,weighing standard and glassware tolerance. Standard uncertainty was 0.21% and expanded uncertainty was 0.42%. The range of content determination was (99.76 ± 0.42)%,k=2. CONCLUSIONS:The method is suitable for uncertainty evaluation of Lamivudine tablets by HPLC,and provides reference for the quality control in laboratory and objective evaluation of Lamivudine tablets.

Objective To evaluate the impact of the recombined plasmid vector with enhanced green fluorescent protein (EGFP) encoding soluble tumor necrosis factor related apoptesis inducing ligand (pIRES-EGFP-sTRAIL) on proliferation and apoptosis of human hepatocellular carcinoma cell line HepG2, and investi-gate the feasibility and efficiency of the transfection of pIRES- EGFP- sTRAIL into HepG2 by ultrasound micro-bubble contrast agent. Methods pIRES-EGFP-sTRAIL was constructed and transfected into HepG2 cells by using different types of mediated methods: microbubble echocontrast agent combining appropriate dose of ultra-sound irradiation, liposome method, microbubble echocontrast agent only or blank medium treatment. Transfec-tion efficiency was evaluated by EGFP-expressed cell count; proliferation-lnhibiting rate and the apoptosis rate of HepG2 cells were determined by MTT method and flow cytometry analysis; changes of cell morphology were examined by microscopy with Hoechst33258 dyeing; expression of caspase-8 and caspase-3 was detected by Western blot. Results Ultrasound microbubbh enhanced pIKES-EGFP-sTRAIL uptake by HepG2 cells, and the transfection efficiency was significantly higher in ultrasound microlmbble group than that in other groups( P<0.05 ) ; pIRES- EGFP- sTBAIL effectively inhibited HepG2 cell proliferation and induced cell apoptosis by triggering caspase cascade. Both the inhibiting rate and apoptosis rate were significantly higher in ultrasound microbubble group than those in other groups(P<0.05). Conclusion pIRES-EGFP-sTRAIL expresses ef-fectively in HepG2 cells, sTRAIL has a potential role on the inhibiting proliferation and inducing apoptosis of HepG2 cells by triggering caspase cascade, and this role can be enhanced by the administration of low-intensity ultrasound and microbubble echecontrast agent.

Objective To report three cases of fulminant ulcerative colitis (FUC) treated with infliximab (IFX) and the positive result for clinical purpose. Methods Three patients with FUC were infused with IFX (5 mg/kg) at interval of 0, 2, 6 wk. Sulfasalazine or probiotics was used for the maintanance of remission. The mucosal healing was evaluated by endoscopy and patholoic examination. ResultsComplete remission was found in 3 patients with FUC. Eight weeks after IFX withdraw, complete remission was found in two of three cases demonstrated by endoscopy, except for one case died from refractory ventricular tachycardia. The remaing 2 cases showed no active manifestation during 8 months' follow-up. Conclusion IFX therapy results in complete remission in 3 cases with FUC. However, further randomized control study is warranted for concrete evaluation on salty and application clinically.

Objective To evaluate the diagnostic value of computed tomography enteroclysis/enterography (CTE),magnetic resorance enteroclysis/enterography (MRE) and ultrasonography (US) in Crohn's disease (CD) by Meta-analysis.Methods Segmental intestinal wall thickening and strengthening was taken as image diagnostic indicators in CD.Endoscopic and histopathologic findings and follow-up results were set as diagnostic standard.Retrieve and literature inclusion criteria were developed.The papers of CD diagnosed by CTE,MRE and US which met the criteria were searched and screened.The quality of the papers was evaluated by quality assessment of diagnostic accuracy studies (QUADAS).The diagnostic efficacy of CTE,MRE and US was compared,which included sensitivity,specificity,positive likelihood ratio and negative likelihood ratio,positive predictive value (PPV) and negative predictive value (NPV).Results Among the 2197 retrieved literature,a total of 12 papers met the criteria and nine of which were blinded study.The papers were grouped by three different imaging techniques and underwent QUADAS scale scoring.One paper was about CTE and MRE comparative study,so there were six papers in CTE group,the scores of all the six papers were over 10 and all of them were blinded study.There were four papers in MRE group,the scores of three papers were over 10 and all of them were blinded study.There were three papers in US group,the score of one paper was over 10 and two of these three papers were blinded study and one with unclear evaluation.The sensitivity of CTE group was the highest (89 ％),while the specificity (90 ％) was lower than that of US group (95％).The PPV (93％),NPV (95％),positive likelihood ratio (15.16) and negative likelihood ratio (0.17) of US group were the highest,those of CTE were secondary (PPV 91％,NPV 82％,positive likelihood ratio 6.25 and negative likelihood ratio 0.15).The sensitivity (74％),specificity (84％),PPV (80％),NPV (80％),positive likelihood ratio (4.18) and negative likelihood ratio (0.33) of MRE group were lower than both US group and CTE group.Conclusions The sensitivity of CTE in CD diagnosis is better than that of MRE and US,while other diagnostic indicators are poorer than US.Even though US has high diagnostic value in CD,more well designed large sample study were still needed for further evaluation.

OBJECTIVETo investgate the expression patterns of bone morphogenetic protein-2 (BMP-2) in the distraction area following plRES-hBMP2-VEGFI65 gene transfection at different time during mandibular distraction osteogenesis in a rabbit model.

METHODS48 New-Zeland rabbits were employed to underwent osteotomy and distraction devices implantation on mandible bilaterly. After 3 days of latency period, the devices were activated at the rate of 0. 8 mm per day for 10 days. The rabbits were randomly divided into 4 groups (group A, B, C and D). Group A, B and C were transfected recombinant plasmids pIRES-hBMP2-hVEGF165 via electroporation-mediated approach at latency period, distraction period, consolidation period respectively. Group D was used as control group without gene transfection. Three rabbits in each group were sacrificed at 1, 2, 4 weeks of consolidation respectively. The mandibles were harvested for immunohistochemical staining detection of BMP-2 expression respectively, which were analyzed by CMIAS series multifunction color quantitative analysis of pathological image analysis system.

RESULTSBMP-2 expression was found to be mainly located in the monocyte, fibroblast of the granulation tissue, the osteoblasts, osteocyte on the surface of new formed trabecular, and the connective tissues surrounding the new formed bone. The expression in group B was superior to other groups. Image analysis showed that, at the first week and second week of consolidation, the expression abosordbance A in group B (0. 58 ± 0. 03 and 0. 34 ± 0. 02) was relatively higher, when compared with that in group A (0. 42 ± 0. 02 and. 31 ±0.01), C(0.32 ±0.01 and 0.30 ±0.01)and D(0.27 ±0.01 and 0.23 ±0.02), showing a significant difference(P <0. 05). It was also relatively higher in group A(0. 42 ± 0. 02 and 0. 31 ± 0. 01) and C(0. 32 ± 0.01 and 0.30 ± 0.01), when compared with that in group D(0. 27 ±0.01 and 0.23 ± 0. 02), showing a significant difference( P < 0. 05) , but there was no significant difference ( P > 0. 05) between group A and group C. At the fourth week of consolidation, the expression decreased and there was no significant difference among group A, B, C, D.

CONCLUSIONSThe electroporation-mediated gene transfection which is transfected at the beginning of traction can promote BMP-2 expression effectively, stimulate bone marrix synthesis and induce proliferation and differentiation of fibroblasts, osteoblasts, endothelial cells, which further effectively promote the new bone formation. It suggests that the distraction stage is the optimal time for gene therapy.

Animals ; Bone Morphogenetic Protein 2 ; metabolism ; Electroporation ; methods ; Genetic Therapy ; methods ; Mandible ; surgery ; Osteogenesis ; Osteogenesis, Distraction ; Osteotomy ; Rabbits ; Time Factors ; Transfection ; methods

[Objective]To explore the clinical experience of the treatment of coronary heart disease by Professor TANG Shuhua.[Methods]Through the clinical study with the teacher to collect and collate the relevant information and medical records, from the etiology and pathogenesis of coronary heart disease, governance and other aspects of the specific analysis and discussion on Professor TANG Shuhua treatment of this disease academic point of view and clinical experience,it summed up the rule of prescription,and to the case of evidence,through regular follow-up,observed the treatment effect. [Results]Professor TANG Shuhua believes that"Yin deficiency endogenous heat and blood stasis" is an important pathogenesis of coronary heart disease. Professor TANG in the treatment of coronary heart disease takes the pathogenesis as a guide, with the proposed nourishing liver and kidney, Qingrejiedu, invigorates the circulation of general pulse function applied to add and subtract, on the basis of Lu Huang Ke Li significantly improves the symptoms of coronary heart disease angina pectoris,delays the development of coronary heart disease progression and improves the quality of life.[Conclusion]Professor TANG Shuhua's treatment of coronary heart disease by nourishing Yin and heat and activating blood circulation is significant, which is of great significance to guiding the treatment of coronary heart disease.

This study is aimed to design a chemically-defined serum free medium (CDSFM) to support in vitro culture of bone marrow-derived mesenchymal stem cells (BM-MSCs). BM-MSCs were isolated from the femoral bones of one-month-old New Zealand Rabbits with density gradient centrifugation. We compared the proliferation capability, cell cycle, colony-forming efficiency, osteogenic and adipogenic differentiation capabilities of BM-MSCs cultured in CDSFM with those cultured in serum-containing medium (SCM). After 10 days culture, BM-MSCs were expanded by 50 folds in CDSFM, while only 40 folds in SCM. EGF, bFGF and hy-drocortisone were the most important additives and significantly stimulated BM-MSCs proliferation. The percentage of cells at G0-G1 cell cycle was 80.31% +/- 0.58% after CDSFM culture, with no significant difference (P>0.05) compared to 75.24% +/- 4.05% for SCM culture. However, the cloning efficiency of BM-MSCs cultured in CDSFM was significantly lower than that in SCM (P<0.01). The expanded BM-MSCs in CDSFM preserved differentiation potentials into mesenchymal lineages in vitro, including adipocytes and osteoblasts. We have designed a chemically-defined serum free medium that could support in vitro proliferation and maintain the properties of BM-MSCs as stem cells, which could be applied to cell-based therapy and biomedical research.
Animals ; Bone Marrow Cells ; cytology ; Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Culture Media, Serum-Free ; Mesenchymal Stromal Cells ; cytology ; Rabbits

Objective:To investigate the effects of CT images reconstructed using different field of view (FOV) sizes on the automatic segmentation of organs at risk and dose calculation accuracy in radiotherapy after radical mastectomy.Methods:Under the same scanning conditions, CT values-electron density conversion curves were established by reconstructing the original CT images of a phantom placed at the isocenter and extended FOV (eFOV) positions using FOV sizes of 50, 60, 70 and 80 cm. Then, these curves were compared. A standard phantom with a known volume was scanned, and the automatic segmentation result of the phantom on CT images reconstructed using different FOV sizes was compared. A total of 30 patients in Guangdong Second Provincial General Hospital from January 2020 to June 2022 with breast cancer were randomly selected. Through simulated positioning, their CT images were reconstructed using different FOV sizes for the purpose of automatic segmentation of organs at risk, followed by comparison between the outcomes of automatic segmentation and physicians′segmentation. The treatment plan established based on CT images reconstructed using a FOV size of 50 cm (FOV 50 images for short) was applied to CT images reconstructed using FOV sizes of 60, 70 and 80 cm (FOV 60, FOV 70 and FOV 80 images for short) for dose calculation, and the dose calculation result were compared. Results:The CT values - electron density conversion curves derived from CT images reconstructed using different FOV sizes were roughly consistent. At the isocenter, the difference between the segmented volume and actual volume of the standard phantom increased up to a maximum of 6 cm 3 (4.8%) with an increase in the FOV size. As indicated by the automatic segmentation result, the segmentation accuracy of the spinal cord, trachea, esophagus, thyroid, healthy mammary gland, and skin decreased with an increase in the FOV size ( t = -28.43-8.23, P < 0.05). The comparison of dose calculated based on CT images reconstructed using different FOV sizes showed that there was no statistically significant differences( P>0.05) in the dose to target volume ( V95) and the maximum and average doses in the supraclavicular lymph node region, as well as the dose to organs at risk. The coverage for planned target volume decreased with an increase in the FOV size, with a maximum difference of 4.06%. Conclusions:It is recommended that, for radiotherapy after radical mastectomy, FOV 50 images should be selected for the automatic segmentation of organs at risk, CT-values-electron density conversion curves should be established based on the electron density phantom images of the eFOV region, and the eFOV 80 images should be preferred for dose calculation.

Objective:To investigate the effect of eyebrow arch augmentation with personalized polyether ether ketone (PEEK) implant assisted by endoscope.Methods:From January 2019 to August 2021, 9 patients with low and flat eyebrow arch requiring eyebrow arch augmentation were treated in the plastic surgery department of the Friendship Plastic Surgery Hospital Affiliated to Nanjing Medical University. The cranial and maxillofacial CT scanning data were obtained before operation, the eyebrow arch elevation range and height were designed according to the patient's requirements, and personalized peek implants were manufactured. During the operation, a small temporal hairline incision was used to accurately separate the implantation cavity of individualized peek implants under the periosteum of the eyebrow arch implantation area with the aid of endoscope. Personalized PEEK implants were implanted under endoscope to make them fully fit with the eyebrow arch bone and fixed with titanium nails. Photos were taken and analyzed before and after operation.Results:All patients were followed up for 3 months to 2 years. Their facial expressions were normal, and there were no motor nerve injury, rejection and iatrogenic infection. The patients were satisfied with the postoperative effect. Compared with before operation, the low level of orbital bone in eyebrow area was improved, and the facial contour was three-dimensional and harmonious.Conclusions:The eyebrow arch augmentation with individualized peek implant assisted by endoscope is safe and effective. It can greatly reshape the upper contour of the face, deepen the upper eyelid fossa and improve the convex eye phenomenon, so as to achieve the cosmetic effect of enhancing the three-dimensional sense of the eyebrow orbital area, which is worthy of clinical application.