[Effects of Trunk Control Training on Dynamic Sitting Balance and Trunk Function in Hemiplegia Patients after Acute Stroke].

OBJECTIVE: To investigate the effect of trunk control training on trunk function, balance, mobility and functional independence in hemiplegia patients after acute stroke, and to compare the therapeutic effects with conventional stroke rehabilitation. METHODS: The study was designed as a randomized controlled trial. The subjects and evaluators were blinded and the therapist was unblinded. Subjects were randomly assigned to either the experimental group or the control group using a computer-generated random number table, the experimental group ( n=15) received30 min of high-intensity trunk control training plus 15 min of low-intensity conventional stroke rehabilitation (45 min, once1 d for 5 d). The control group ( n=15) received only 45 min low-intensity conventional stroke rehabilitation, once 1 d for 5 d. Trunk function was the primary outcome, which was evaluated by the Trunk Impairment Scale (TIS). The secondary outcome included balance, mobility and functional independence, which were assessed by the Brunel Balance Assessment (BBA), the Modified Rivermead Mobility Index (MRMI) and the modified Barthel Index (MBI). Subjects were assessed before and after the intervention. RESULTS: After 5 d of treatment, the scores of TIS general table, TIS static sitting balance sub table, TIS coordination sub table, BBA, MRMI and MBI were increased, but there was no statistically significant difference between the two groups (all P>0.05). The trunk control training group improved dynamic sitting balance more significantly, the score of TIS dynamic sitting balance sub table increased more than that in the control group, with a statistically significant difference ( P<0.05). CONCLUSION: The results of this study indicate that trunk control training has improved trunk function, balance, mobility, and activities of daily living in hemiplegia patients after acute stroke, and trunk control training improves dynamic sitting balance more effectively than conventional stroke rehabilitation.

[Semen quality and HIV RNA level in HIV/AIDS male patients after treated by highly active antiretroviral therapy].

OBJECTIVE: To evaluate the semen quality of the HIV/AIDS male patients after treated by the highly active antiretroviral therapy (HAART) and their potential of transmitting HIV/AIDS and provide some evidence for this cohort of males who wish for parenthood. METHODS: We collected semen samples from 20 HIV/AIDS male patients who had been treated by HAART for over 6 months and wished for parenthood. We examined sperm concentration, viability and total motility and the percentage of morphologically normal sperm (MNS) using the computer-assisted semen analysis system, measured the HIV-1 RNA loads in the semen by the Cobas Amplicor Monitor test, and counted CD4+ T cells in the peripheral blood by flow cytometry. RESULTS: The patients were aged 25－40 (30.7 ± 5.05) years. After treated by HAART for 6－26 (14.24 ± 12.26) months, the count of blood CD4+ T cells was significantly increased (341－1 058 ï¼»535.76 ± 212.021ï¼½ /µl) in comparison with the baseline (226－965 ï¼»422.38 ± 200.86ï¼½ /µl). Compared with the normal value, the semen volume was increased except in 1 case (≥2 ml) while total sperm motility was decreased in 13 cases (≥40%), and so were sperm concentration in 2 cases (≥15 × 106 / ml), sperm viability in 5 (58%), the percentage of progressively motile sperm in 18 (≥32%), and the percentage of MNS in 6 (≤4%). HIV-1 RNA in the peripheral blood was <20 copies/mL in all the cases and that in the seminal plasma was also <20 copies/ml in 18 cases but >20 copies/mL in the other 2 (ï¼»4.70 × 101ï¼½ and ï¼»2.2 × 102ï¼½ copies/ml, respectively). Of the 4 couples that had sex without protective measures for over 6 months, all the 4 female partners exhibited negative HIV antibodies in regular follow-up examinations and 1 achieved spontaneous pregnancy and healthy birth, with negative HIV-1 RNA in both the mother and the baby. CONCLUSIONS: The HIV RNA level is higher in the semen than in the blood of the HIV/AIDS male patients after HAART, which indicates the potential risk of their semen transmitting HIV/AIDS to their female partners. Their sperm concentration and total sperm motility are lower than the normal value, which suggests a decreased fertility.

[The ancient literature source and flow of the point to point penetration acupuncture method].

It has been the consensus of the academic community that the doctor DOU Han-qing in Jin-Yuan Dynasty was the pioneer of point to point penetration acupuncture method.The contents of several texts in Yuan-Ming-Qing Dynasties were combed on the basis of Dou Taishi Zhenjing(《》).The paradigm,the acupoints,basic elements and the characteristics of the main treatment of diseases were discussed,and the relationship between the point to point penetration acupuncture method with the penetration acupuncture method has been tried to distinguish from the perspective of technical philosophy. It was found that the specifications and quantity of the 28 "penetration and to" cases in Dou Taishi Zhenjing(《》)were sufficiently exemplary for future generations, and were the source documents of Yuan-Ming-Qing's texts,which basically followed its specifications and content and developed.For example,the Yulong Poetries(《》)in the books of Bianque Shenying Zhenjiu Yulong Jing(《》),Zhenjiu Dacheng(《》),Zhenfang Liuji(《》).The 37 cases in Zhenfang Liuji(《》) were the most in several texts.The 31 cases in Xunjing Kaoxue Bian(《》)had the characteristics of DOU Han-qing, YANG Ji-zhou and LING Yun, who developed the opposite direction penetration method with two or more needles on DOU Han-qing's one needle.There were five basic elements in the point to point penetration acupuncture method:the needle pri-cking inch(depth), the needle pricking angle, the needle pricking direction, the starting acupoint, the end acupoint.This method in Dou Taishi Zhenjing(《》)mostly treated local diseases and symptoms,some acupoints below the elbows and knees also treated far-end diseases.The point to point penetration acupuncture method has a specific intention structure, which cannot be confused with the penetration acupuncture method.

[Exploration of the construction of semantic framework of meridians and acupoints based on top-level ontology].

Based on the top-level ontology and the existing ontology methodology, the related concepts of meridians and acupoints were discriminated, defined and classified; the relationship of core concepts were established, e.g. meridians, acupoints and zangfu. It was attempted to build an ontological semantic framework of meridians and acupoints. Through the investigation on the classification mode of the top-level ontology, it is proposed that the meridians and acupoints, as the unique concepts of traditional Chinese medicine, exist in the form of "emptiness" and belong to "immaterial entity". Meridians refer to the three-dimensional channels in the human body, and acupoints are divided into ontological acupoints and body surface ones. Ontological acupoints are regarded as a three-dimensional structure within the human body, whereas, body surface ones are the optimal sites for acupuncture needle insertion on the body surface, meaning, the zero-dimensional point on the body surface. The main relationships between meridians and acupoints include is-a, exterior-interior, located-in, correspondent-to, mapping, etc. The exploration of the semantic framework of meridians and acupoints is conductive to understanding the connotation of meridians, acupoints and their relationship.

The optimum parameters and neuroimaging mechanism of repetitive transcranial magnetic stimulation to post-stroke cognitive impairment, a protocol of an orthogonally-designed randomized controlled trial.

OBJECTIVE: Repetitive Transcranial Magnetic Stimulation (rTMS) has been used in cognition impairment due to various neuropsychiatric disorders. However, its optimum parameters and the neuroimaging mechanism are still of uncertainty. In order to simulate a study setting as close to real world as possible, the present study introduces a new orthogonally-designed protocol, consisting of the rTMS intervention with four key parameters (stimulating site, frequency, intensity and pulse number) and three different levels in each one, and aims to investigate the optimum parameters and the brain activity and connectivity in default mode network (DMN), dorsal attention network (DAN), central executive network (CEN) following rTMS intervention to post-stroke cognition impairment (PSCI). METHODS: A single-center, orthogonally-designed, triple-blind randomized controlled trial will be conducted and forty-five PSCI patients will be recruited and randomly assigned to one of nine active rTMS groups based on four rTMS paraments: stimulating site, frequency, intensity and pulse number. Neuropsychological, activities of daily living, quality of life and functional magnetic resonance imaging (fMRI) evaluations were be performed pre-, post- and 3 months after rTMS. DISCUSSION: This study evaluates the optimum parameters of rTMS for patients with post-stroke cognition impairment and explores the alteration of neural function in DMN, DAN, CEN brain network. These results would facilitate the standardized application of rTMS in cognition impairment rehabilitation.

Cerebellar Theta Burst Stimulation on Walking Function in Stroke Patients: A Randomized Clinical Trial.

Objectives: The objective of this study was to explore the efficacy of cerebellar intermittent theta burst stimulation (iTBS) on the walking function of stroke patients. Methods: Stroke patients with walking dysfunction aged 25-80 years who had suffered their first unilateral stroke were included. A total of 36 patients [mean (SD) age, 53 (7.93) years; 10 women (28%)] were enrolled in the study. All participants received the same conventional physical therapy, including transfer, balance, and ambulation training, during admission for 50 min per day during 2 weeks (10 sessions). Every session was preceded by 3 min procedure of cerebellar iTBS applyed over the contralesional cerebellum in the intervention group or by a similar sham iTBS in control group. The groups were formed randomly and the baseline characteristics showed no significant difference. The primary outcome measure was Fugl-Meyer Assessment-Lower Extremity scores. Secondary outcomes included walking performance and corticospinal excitability. Measures were performed before the intervention beginning (T0), after the first (T1) and the second (T2) weeks. Results: The Fugl-Meyer Assessment for lower extremity scores slightly improved with time in both groups with no significant difference between the groups and over the time. The walking performance significantly improved with time and between group. Two-way mixed measures ANOVA showed that there was significant interaction between time and group in comfortable walking time (F 2,68 = 6.5242, P = 0.0080, η2 partial = 0.276, ε = 0.641), between-group comparisons revealed significant differences at T1 (P = 0.0072) and T2 (P = 0.0133). The statistical analysis of maximum walking time showed that there was significant interaction between time and groups (F 2,68 = 5.4354, P = 0.0115, η2 partial = 0.198, ε = 0.734). Compared with T0, the differences of maximum walking time between the two groups at T1 (P = 0.0227) and T2 (P = 0.0127) were statistically significant. However, both the Timed up and go test and functional ambulation category scale did not yield significant differences between groups (P > 0.05). Conclusion: Our results revealed that applying iTBS over the contralesional cerebellum paired with physical therapy could improve walking performance in patients after stroke, implying that cerebellar iTBS intervention may be a noninvasive strategy to promote walking function in these patients. This study was registered at ChiCTR, number ChiCTR1900026450.

[Collection methods, stylistic layout and value of extraordinary points in Doutaishi Zhenjing (Master Dou's Canon of Acupuncture)].

The literature source, collection methods and stylistic layout, value and influence of 32 extraordinary points in Doutaishi Zhenjing (Master Dou's Canon of Acupuncture) were collected. The sources of the literature are extensive. The collection methods include 5 aspects, named inheritance, integration and repair; name rectification, identification and differentiation; new point explication on the base of extraordinary points; well arrangement of points and mutual use of three methods for stylistic layout; as well as integrative illustration with text and picture, and determined location. In reference to the holistic layout of name, location, techniques of acupuncture and moxibustion and indications of extraordinary points in Doutaishi Zhenjing (Master Dou's Canon of Acupuncture), it is the first time to make a standardized and unified collection of extraordinary points, which is valuable in the historical literature, the theoretic research and clinical practice of extraordinary points. It brings a great impact on the special articles or books after the Yuan dynasty and even the establishment of national standards at present.

[Relationship between DOU Han-qing of the Jin and Yuan Dynasty and Taoist].

In order to explore the culture factors of the construction of acupuncture theory of DOU Han-qing, the medical master of the Jin and Yuan Dynasty, the authors studied the relationship between DOU Han-qing and Taoist by collecting DOU 's life material. With manual retrieval, some ancient historical literature was obtained. Additionally, Yijing Huiyuan: Zhenjiu Yuanshu: Dou Taishi Michuan Mihua Zhenjing Qiongyao Baojian, written by WU Jia-yan in the Ming Dynasty, collected in Japan, was analyzed. It was found that DOU Han-qing's acupuncture and moxibustion was influenced by Suwen Wangpian: Cifalun, the Taoism medicine of the early stage of the Song Dynasty, as well as by Jiaojing Baxue, passed on by SONG Zi-hua, the hermit. DOU Han-qing had been in contact with FAN Zhi-ying of the Taoist, but there was no clear record relevant with medicine. Dou Taishi Michuan Mihua Zhenjing Qiongyao Baojian may be the compilation by Taoism medical master, on the base DOU 's acupuncture and moxibustion. There is a kind of mutual influence and mutual promotion relationship between DOU Han-qing and Taoist.

[Objective, approach and symbolic achievement of the standardization on extraordinary points].

Through sorting out the development of meridian points and the standardization of extraordinary points in dynasties, two objectives are discovered in the standardization of extraordinary points starting from the Tang Dynasty. The points with extraordinary effect are selected points and transferred to meridian points and the commonly-used key points are selected and developed to be standardized acupoints. Hence, 3 approaches are summarized. Approach No.1: a large number of "meridian-outside" points that are the potential supplementation but not included yet in meridian points are supplemented to meridian points. Approach No.2: based on the specified requirement, the key points are selected from the acupoints with determined locations and names, which is developed to be the standard of acupoints. Approach No.3: the acupoints with extraordinary effect are selected from the "meridian-outside" points and these points are collected for the formulation of the standard of extraordinary points. The latter two approaches provide for the development direction of teaching and standardization in the future.

[Evolution of the concept of extra ordinary point and standardization of related terms].

Through an analysis of the historical development of the concept of extra ordinary point, it is found that the terms of "Qi point" "extra ordinary point" "Loujing point" "extra meridian point" and "Bie point" used in the ancient medical books in the Yuan, Ming, and Qing Dynasties actually expressed two different concepts of "extra ordinary points with the magical effects" and "extra points other than the meridian points". The standardized term for the former concept should be "extra ordinary point" (Qi point for short), and the standardized term for the latter concept should be "extra meridian point", which is also called "Loujing point" and "Bie point". Since these two different concepts are not clarified in modern acupuncture textbooks, especially the misreading of "Jing Wai", no consensus has been reached on the definition of "extra ordinary point" and "extra meridian point", and the standardized terms have not been decided, which changed from "extra ordinary point" to "Qi point" and "extra point" and recently returned to "extra ordinary point". These confusions in textbooks have a direct impact on the determination of the related acupuncture terms used in national and international standards.

[Overview of studies on DOU Han-qing's academic thoughts about acupuncture-moxibustion].

DOU Han-qing (1196-1280 A．D.) is a famous acupuncturist of the Jin and Yuan dynasties in ancient China. His academic thoughts about acupuncture and moxibustion have a higher clinical practical and theoretical value and received extensive attention in China. In the present paper, we make systematic analysis and summary about the current researches on Master DOU's academic thoughts after retrieval and screening the related research literature (published from the foundation of the People's Republic of China till now) by using electronic databases. The results showed that our understanding about Master DOU's academic thoughts are mainly based on Zhenjing Zhinan (《》，A Guide to the Classic of Acupuncture), especially the Biaoyou Fu (《》，Lyrics of Recondite Principles) and the eight influential acupoints of meridians, but less attention has been paid to books DOU Taishi Zhenjing (《》，Master DOU's Acupuncture Classics), Zhenjiu Jicheng (《》，Compendium of Acupuncture-moxibustion) and Panshijin Zhici Michuan (《》，Panshijin Esoteric Techniques on Perpendicular Insertion of Acupuncture Needles). The systematicness and depth of research on Master DOU's academic theory need being improved. We propose that we should make a comprehensive study about Master DOU's books and his life stories under the academic development history of overall background of Chinese acupuncture and moxibustion, so as to get a clear, complete and objective understanding on DOU's academic thoughts.

Induction of specific cytolytic T lymphocytes using fusions of hepatocellular carcinoma (HCC) patient-derived dendritic cells and allogeneic HCC cell line.

BACKGROUND/AIMS: To assess the ability of hepatocellular carcinoma (HCC) patient-derived dendritic cells (DCs) fused with allogeneic HCC cell line to activate autologous lymphocytes to generate specific cytotoxic T lymphocytes (CTL) in vitro. METHODOLOGY: DCs were obtained by culturing adherent peripheral blood mononuclear cells (PBMC) from HCC patients in the presence of 100 microg/L recombinant human granulocyte/ macrophage- colony stimulating factor (rhGM-CSF) and 20 microg/L interleukin-4 (rhIL-4) for 1 week in vitro. DCs were fused with allogeneic HCC cell line HepG2 cells using polythyleneglycol (PEG), and the fusion cells were designated as DCs/HepG2. By labeling DCs and HepG2 with green and red fluoresceins, respectively, the cellular fusion was examined under fluorescence microscope. The ability of DCs/HepG2 to stimulate proliferation and differentiation of autologous lymphocytes was assessed by MTT method, and the specific killing efficacy of DCs/HepG2-induced CTL against HepG2 was evaluated. RESULTS: HCC patient-derived DCs expressed a certain level of CD1a, HLA-DR, CD54, CD80 and CD86. Fluorescence microscopic examination demonstrated that co-incubation of DCs and HepG2 in the presence of PEG lead to generation of DCs/HepG2. In the mixed lymphocyte reaction assay, DCs/HepG2 had a significantly greater ability to activate proliferation of autologous lymphocytes, as compared with DCs alone, DCs plus HepG2, HepG2 alone and medium control (P<0.05). The DCs/HepG2-activated CTL showed a potent specific killing efficacy against HepG2 cells. CONCLUSIONS: Fusions of HCC patient-derived DCs and allogeneic HCC cell line could efficiently stimulate autologous lymphocytes to generate tumor-specific CTL in vitro. It might represent a promising approach of immunotherapy for HCC.

Anesthetic agent etiomidate induces apoptosis in N2a brain tumor cell line.

The present study identified the cytotoxic effects of etomidate on the N2a neuroblastoma cell line. Etomidate induced apoptosis in N2a cells in a concentration­dependent manner, which was confirmed by western blotting and flow cytometry. Phase contrast microscopy was used to analyze the effect of etomidate on morphological characteristics. The number of the apoptotic cells was increased and confirmed by DAPI and PI staining, which served as a characteristic hallmark of apoptosis. Additionally, etomidate led to loss of mitochondrial membrane potential and resulted in the generation of reactive oxygen species in N2a cells. The western blot analysis revealed that N2a cells treated with etomidate had a significant modulation of pro­apoptotic proteins, includingpoly ADP­ribose polymerase (PARP), cleaved PARP, caspase­9 and procaspase­3. In conclusion, the present study determined that etomidate induced cytotoxic and apoptotic effects in N2a brain tumor cells in vitro.

Vascular damage and anti-angiogenic effects of tumor vessel-targeted adenovirus-mediated herpes simplex virus thymidine kinase gene.

AIM: To explore the therapeutic efficacy and mechanism of herpes simplex virus-thymidine kinase (HSV-tk) targeting angiogenesis against hepatocellular carcinoma in vivio and in vitro. METHODS: Recombinant adenovirus containing kinase domain insert with receptor (KDR) or cytomegalovirus (CMV) promoter-controlled HSV-tk gene (AdKDR-tk and AdCMV-tk) was constructed using pAdeasy system. The expression of KDR antigen in human umbilical venous endothelial cells (HUVEC) and HepG2 was detected with histological analysis of cells. The virus was used to infect HUVEC and HepG2. Following administration of ganciclovir (GCV), the survival rate of gene-transfected HUVEC and HepG2 was evaluated by MTT method. To develop hepatocarcinomas in 32 Balb/C mice with HepG2 cells, the mice were divided into four groups: ganciclovir group (I), Ad group (II), AdCMV-tk group (III) and AdKDR-tk group (IV). Then selective administration of recombinant adenovirus or Ad via the intratumorial was given to all rats. Ganciclovir (GCV) was given at a dose of 100 mg kg(-1) d(-1) (ip) started on the following day and lasted 10 d. Microvessel density (MVD) of tumor in all the treated animals were examined by the immunohistochemical methods and tumor burden was evaluated 10 d before and after the last GCV dose. RESULTS: Immunocytochemical staining indicated the expression of KDR antigen in HUVEC. Under adenovirus infection index of 100, with increasing GCV concentration from 0 up to 50 mg/L, the survival rate of AdKDR-tk-transfected HUVEC and HepG2 decreased from 100% to (28.94 +/- 5.67)% and (75.45 +/- 2.91)% at proper order, respectively (P < 0.01), while the survival rate of AdCMV-tk-transfected HUVEC and HepG2 declined from 100% to (17.56 +/- 2.48)% and (23.15 +/- 5.72)%, respectively (P > 0.05). Compared with group I, there was a decrease of tumor weight by 14.7% in group III and by 23.6% in group IV. And there was a distinct difference between group III and IV (P < 0.05). The median MVD for all groups was 37.4 +/- 8.6, 30.6 +/- 7.8, 27.6 +/- 7.1, and 10.7 +/- 4.1 (microvessels/mm(2)) in group I, II, III and IV, respectively. And there was a marked difference between group III and II (P < 0.05), IV and II (P < 0.01), and IV and III (P < 0.01). CONCLUSION: KDR promoter-HSV-tk gene may effectually restrain the growth of tumor via targeting angiogenesis for hepatocellular carcinoma with treatment of GCV.

[Remodel retinal neovascularization by overexpression of HESR-1 in vitro].

OBJECTIVE: To remodel retinal neovascularization by overexpression of HESR-1 in vitro. METHODS: Human retinal capillary endothelial cell (HRCEC) was isolated, then HRCECs were transfected with pcDNA3.1+HESR-1. The changes of expression of KDR and occluding in HRCECs were analyzed using ELISA assay, the regulation of occluding in HRCECs by HESR-1 was evaluated by western blot. RESULTS: Over-expression of HESR-1 down-regulated KDR expression, but occludin expression was up-regulated in HRECEs, the result also showed that the vessels permeability was reduced by HESR-1. CONCLUSIONS: Overexpression of HESR-1 in HRCECs can inhibit the neovascularization and decrease the permeability of new vessels. It may be a promising gene therapy for intraocular neovascularization.

[Study on the origin of Dou Han-qing's academic thought of acupuncture and moxibustion].

The origin of Dou Han-qing's academic thought of acupuncture and moxibustion is studied by analysis on relative contents in the books Weisheng Baojian, Jisheng Bacui and Zhenjiu Sishu Zhenjing Zhinan and the two papers "On reinforcing-reducing manipulation" and "Questions and answers of qi and blood", holding that DOU Han-qing's academic thought of acupuncture and moxibustion is influenced more by Suwen (Plain Questions) and Nanjing (Classic on Medical Problems), not by Lingshu (Miraculous Pivot).

[Textual research and differentiation of Dou Han-qing's works catalogue].

This paper aims at probing into evolving course of DOU Han-qing's works catalogue. On the basis of summarizing and referring to study achievements of our predecessors, through analysis of book lists and relative works and chapters, it is hold that the catalogue which were not attained by ZHULiang-neng possibly are the contents of acupuncture reinforcing and reducing methods; the books printed and published by ZHULiang-neng include the contents of both channels and acupoints; the book, Zhinan, which was attained by DOUGui-fang, includes the content catalogue of needling methods; Fu Zhenjiu Zashuo in Zhenjiu Sisu. Zhenjiu Zhinan also were extracted by DOU Gui-fang from Illustrated Manual of Acupoints of the Bronze Figure, and The Zhenjiu Biji Taiyi Zhi Tuxu and Dongzhi Yezhe Gongshuo should belong to The Fu Zhenjiu Zashuo.

[Construction and identification of recombinant vectors carrying herpes simplex virus thymidine kinase genes expressed in vascular endothelial cells].

BACKGROUND & OBJECTIVE: Blocking tumor angiogenesis is a promising tumoricidal approach. Herpes simplex virus-thymidine kinase (HSV-tk) suicide gene could effectively damage neoplastic vascular endothelial cells. However, the killing effect of currently used HSV-tk system is non-specific due to employing cytomegalovirus (CMV) as its promoter. Kinase domain insert containing receptor (KDR), a kind of vascular endothelial growth factor receptor, is highly expressed on tumor vascular endothelial cells, but lowly expressed on normal tissues. This study was to construct a recombinant adenovirus containing KDR promoter-mediated HSV-tk suicide gene, and analyze its specific expression on vascular endothelial cells. METHODS: KDR-tk fragment was subcloned into corresponding restriction sites of vector with directional cloning method using pAdeasy system to construct recombinant adenoviral plasmid containing KDR promoter-controlled HSV-tk gene (AdKDR-tk). After packaged and amplified in 293 cells, the virus was used to infect KDR-expressed human umbilical venous endothelial cells (HUVECs) and KDR-unexpressed liver cancer cell line HepG2. After administration of ganciclovir (GCV), proliferation of gene-transfected HUVECs and HepG2 cells was evaluated by MTT assay. RESULTS: According to the results of restriction digestion, reverse transcription-polymerase chain reaction (RT-PCR) and PCR, the length, position and orientation of inserted genes were correct. The titer of the recombinant adenovirus was 1 x 10 (10) pfu/ml. Under infection index of 100, with the concentration of GCV increased from 0 to 50 microg/ml, the survival rates of AdKDR-tk-transfected HUVECs and HepG2 cells decreased from 100% to (28.94+/-5.67)% and (75.45+/-2.91)%, respectively (P<0.01). CONCLUSION: Adenovirus-mediated transfection of KDR promoter-HSV-tk gene could direct endothelial specific gene expression.

Protective effects of transcription factor HESR1 on retinal vasculature.

HESR1 is a basic helix-loop-helix transcription factors regulated by the Notch signaling pathway in vertebrate and Drosophila embryos, and is related to the HES/Hairy/E (sp1) family. HESR1 is a downstream target of Notch in endothelial cells and could be an effector of Notch signaling in these cells. HESR1 is necessary for the induction of a tubular network and for continued maintenance of mature and quiescent blood vessels. To examine the role of HESR1 in retinal neovascularization, we transfected retinal vascular endothelial cells (HRCECs) with the HESR1 gene and studied its effects on the expression of angiogenic factors, on the proliferation and migration of endothelial cells, and on the formation of tube-like structures (TLSs). Overexpression of HESR1 downregulated VEGFR-2 expression, upregulated occludin expression, inhibited the migration and proliferation of HRCECs, and inhibited the formation of TLSs. Thus, HESR1 plays a key role in the finely tuned network of molecules involved in the regulation of retinal vascular homeostasis. HESR1 seems to inhibit the vessel-promoting effects of VEGF, shift endothelial cells from a proliferative state to a quiescent state, and restore normal vessel structures. Expression of the HESR1 gene in retinal vascular endothelial cells may protect retinal blood vessels and may be useful in the treatment of diseases involving damage to the retinal vasculature, including diabetic retinopathy, age-related macular degeneration, and retinal vein occlusion.

[Adenovirus containing c-FLIPs gene induces antiapoptosis activation of lymphocyte in vitro].

OBJECTIVE: To investigate the efficacy and mechanisms of recombinant adenovirus containing c-FLIPs (cellular FLICE-inhibitory protein short) in inducing antiapoptosis activation of lymphocyte in vitro. METHODS: c-FLIPs gene was cloned from total RNA of lymphocyte with RT-PCR methods, then constructed recombined adenovirus Ad.c-FLIPs by pAdeasy system. H9 cells infected with Ad.c-FLIPs produced antiapoptosis derived from anti-Apo-1 antibody, and those antiapoptosis evaluated with PI stain flow cytometry (FCM) and Hoechst stain. RESULTS: c-FLIPs gene was successfully cloned from lymphocytes and incorporated into recombinant adenovirus Ad.c-FLIPs.After exposure to anti-Apo-1 for 24 h, the apoptosis of H9 cells preconditioned by Ad.c-FLIPs for 24 h as measured by Hochst stain or FCM significantly decreased, compared with non-preconditioned cells, 3.60% +/- 0.21% vs. 48.33% +/- 7.41%, respectively, indicating that Ad.c-FLIPs preconditioning protected H9 cells against apoptosis induced by anti-Apo-1. CONCLUSION: Recombinant adenovirus Ad.c-FLIPs effectively induced anti-apoptosis activation of lymphocyte.