Hereditary disease, especially monogenic disease is one of the major causes for malformation and disability of children. Most hereditary diseases have no effective therapy besides clinical symptomatic treatment. Biological techniques targeting casual genes or related signaling genes, such as transgenic, RNA interfere, genome editing, have been successfully applied in treating some hereditary diseases. However, most biological, treatments were carried out in animals, further confirmation of the effectiveness and safety of these therapies, and development of more therapeutic approaches based on mechanisms are needed before clinical trials.
Animals ; Biological Therapy ; Disease Models, Animal ; Genetic Diseases, Inborn ; therapy ; Genetic Therapy ; Humans

Objective To study the effect of cell wall polysaccharide (CWP) of Thallus Laminariae (TL) on thrombosis and blood coagulation.Methods Thrombosis was induced by arteriovenous shunt,ligation of inferior vena cava and electric stimulation of common carotid artery,after intraperitoneal injection of CWP or nomal saline for 3 days.The weight of thrombus and the occlusion time were examined in the normal saline group and high-dose (100 mg/kg) and low-dose (20 mg/kg) groups of CWP of TL.Blood coagulation time (CT),plasma prothrombin time (PT) and partial thromboplastin time (APTT) were measured to observe the effect of CWP on blood coagulation.Results Compared with the control group,both high-dose CWP of TL and low-dose CWP can obviously decrease the weight of thrombus and prolong the occlusion time,CT and APTT.High-dose CWP could also prolong PT obviously,the differences being significant.Low-dosa CWP also prolong PT,but the difference was not significant statistically.Conclusion CWP of TL can inhibit the thrombus formation and blood coagulation in rats.

Objective To improve the efficiency of donated organs and to investigate the possi- bility and operation protocol of heart and lung transplantation from the same donor to different reci- pients.Methods The organs of heart and lung were obtained from three donors through the method of orthotopic lavage and whole cutting,and transplanted to end-stage patients with heart diseases or lung diseases.First,the donor was subjected to mechanical ventilation,thoracotomy and lavage of heart and lung.The heart and lung were cut in whole and separated.The recipients were subjected the re- trograde lavage of heart and lung.In the patients receiving unilateral lung transcription,the left and right lungs were isolated.Among 3 recipients of lung transcription,one underwent left unilateral lung transplantation and 2 bilateral lung transplantation.Three cases received orthotopic cardiac transplan- tation by using bicaval orthotopic heart transplantation (BOHT) anastomosis method.Results One recipient of left unilateral lung transplantation recovered well after operation.In one recipient under- going bilateral lung transplantation,right upper lung embolism occurred.After re-operation of cutting the right upper lung on the 9th day of the first one,he recovered from the illness.In another recipient receiving bilateral lung transplantation,anastomotic stenosis of trathea and pulmonary infection occurred after operation.After expectant treatment,he recovered from the illness,but died from serious infection 7 months after operation.One of the heart recipients was found having renal failure of first stage,and recovered from disease after hemodialysis treatment.Quality of life was good in all 3 heart recipients after operation and their heart function achieved the restoration of 0-Ⅰstage.Conclusion The treatment protocol of transplantation from the same donor to different recipients at the same time, which can make full use of the organs from donors,is feasible and has good effect.

This paper introduces the nursing care for 36 patients with framework exposure in auricular reconstruction by temporal superficial fascia flaps combining with split skin grafts transplantation,such as pre-and post-operative behavioral in-struction,psychological care,and close observation of blood circulation of reconstructed auricular. The patients' wounds healed up well and all the split skin grafts survived successfully. No infection and stent exposure occurred once more. The appear-ance of the reconstructed auricular was pleasing too. It is suggested that high quality of pre-and post-operative nursing,care-ful health education and psychological care be beneficial for the successful repair of framework exposure after auricular recon-struction.

Nanomedicine offers great promise for early diagnosis and treatment of formidable diseases. The unique morphology and biology characteristics of bacteriophage provide unprecedented opportunity for such endeavor. The paper summarizes the application of bacteriophage in nanobiomaterials, nanomedicine, nanomedicine delivery and nanodiagnosis, especially the nano-imaging reagents and future direction concerning nanomedicine based on bacteriophage.

Aim To investigate the roles of intracellu-lar reactive oxygen species ( ROS ) and Nrf2 pathway in shikonin-induced A549 cell apoptosis. Methods The cytotoxicity was analyzed by MTT assay. The ap-optosis of A549 cells was analyzed by both cellular morphological and biochemical methods. The relative changes of the redox marks ( ROS/GSH) were studied by fluorescence assay in the shikonin-treated A549 cells in accompany with the changes of the intracellular redox homeostasis by GSH/GSSG ratio. ROS inhibitor was also employed in the treatment to find the role of ROS in shikonin-induced A549 cell apoptosis. Real-time PCR analysis and ELISA assay were performed as well to determine the role of Nrf2 pathway in the shiko-nin-induced A549 cell apoptosis. Results The IC50 of shikonin on A549 cells was 3. 2 mg·L-1 . The cellu-lar redox homeostasis shifted toward oxidation signifi-cantly in shikonin treatment in a time-dependent man-ner. The expression of the Nrf2 pathway related genes was up-regulated by shikonin ( 3 . 2 mg · L-1 , 8 h ) . The expression of the anti-apoptotic genes was down-regulated , and proapoptotic genes were up-regulated by shikonin (3. 2 mg·L-1, 24h). Futhermore, the inhi-bition of intracellular ROS alleviated the cytotoxicity of shikonin in A549 cells. Conclusion The critical role of shikonin-induced redox imblance in A549 cell, coped with the secondary produced ROS and Nrf2 path-way antioxidants, result in A549 cell apoptosis.

Objective To investigate the effects of 5-azacytidine on radiation-induced epithelialmesenchymal transition in rat alveolar type Ⅱ epithelial cell line (RLE-6TN) and explore their working mechanisms,and to provide experimental evidence for the potential drug-based treatment of radiationinduced pulmonary fibrosis.Methods RLE-6TN cells were cultured in vitro and divided into four groups according to the experimental purposes:control group (C),radiation group (R),5-azacytidine group (A),and radiation followed by 5-azacytidine group (R + A).The microstructural changes in cells were determined by transmission electron microscopy.Inverted phase-contrast microscopy revealed the morphological changes in cells.The mRNA expression levels of E-cadherin and α-SMA were measured by quantitative real-time polymerase chain reaction (qRT-PCR).The protein expression levels of E-cadherin,GSK3 β,and p-GSK3 β (Ser9) were measured by Western blot.The one-way analysis of variance was used for pairwise comparison.Results The cells in group R became spindle-like.Similar morphological changes were not observed in cells in group R + A.Osmiophilic lamellar bodies disappeared at last in cells in group R.RT-PCR results showed that compared with group C,group R had a significantly lower mRNA expression level of E-cadherin ((0.23 ± 0.06) vs.(1.00 ± 0.00),P =0.002)) and a significantly higher mRNA expression level of α-SMA ((2.91 ± 0.01) vs.(1.00 ± 0.00),P =0.000)).However,compared with group R,group R + A had a significantly higher mRNA expression level of E-cadherin ((0.47 ± 0.05) vs.(1.00 ± 0.00),P =0.024)) but a significantly lower mRNA expression level of α-SMA ((2.50 ± 0.02) vs.(1.00 ±0.00),P =0.037)).The results of Western blot showed that the protein expression level of Ecadherin was significantly reduced ((0.07 ± 0.01) vs.(0.48 ± 0.02),P =0.028)),while the protein expression level of p-GSK3β was significantly increased in Group R than in Group C ((0.85 ± 0.04) vs.(0.23 ± 0.03),P =0.031)).However,compared with group R,group R + A had a significantly lower protein expression level of E-cadherin ((0.25 ± 0.00) vs.(0.07 ± 0.01),P =0.024)) and significantly less up-regulation of the protein expression level of p-GSK3β ((0.39 ± 0.03) vs.(0.85 ± 0.04),P =0.014)).Conclusions X-ray radiation can induce the epithelial-mesenchymal transition in epithelial cells.5-azacytidine suppresses radiation-induced epithelial-mesenchymal transition by inhibition of the activity of p-GSK3β in RLE-6TN cells.

Objective: To sieve the bioactive constituents of Radix Puerariae,serum pharmacochemistry research was performed.Method: Based on the establishment of HPLC fingerprints of Radix Puerariae,the constituents absorbed into blood were determined by comparing the HPLC fingerprints of the methanol extracts,tested serum samples and blank serum sample.Results: Four compounds absorbed into blood were detected,among which two were original constituents of Radix Puerariae(including puerarin),the other might be metabolites of the original constituents.Conclusion: These four constituents absorbed into blood were possible bioactive components of Radix Puerariae.Further studies on them will help clarify the bioactive constituents and mechanisms of Radix Puerariae.

To investigate the effect of the anti-endotoxic part of Radix Isatidis on the expression of moesin mRNA in murine tissues induced by lipopolysaccharide (LPS), the sample solution of F(022) part from Radix Isatidis was intraperitoneally administered to experimental mice, and the lipopoly-saccharide (LPS) were injected into the tail vein, and then the tissues of liver, kidney and spleen were colleted and cut into slices. The mRNA was detected by moesin mRNA hybridization in situ. The staining results were observed under microscope. It was found that moesin mRNA expression was increased in the tissues of liver, kidndy and spleen in mice treated with LPS, while in the mice pre-treated with F(022) part from Radix Isatidis, the LPS-induced moesin mRNA expressions in these tissues were inhibited in a dose-dependant manner. Our study showed that F(022) part from Radix Isatidis can inhibit the LPS-induced expression of moesin mRNA in the tissues of liver, kidney and spleen in mice.

Objective:To transfect genes using ultrasound targeted microbubble destruction (UTMD) techniques and observe the effects of RNA interference on cervical cancer (HeLa) cell line in silencing survivin gene and inducing apoptosis. Methods: Recombinant expression plasmid of short hairpin RNA (shRNA) targeting survivin gene was constructed. It was co-treated with microbubbles and transfected to cultured HeLa cells followed by exposure to ultrasound (P+UTMD group). Moreover, blank control group (C), plasmid group (P), ultrasound exposure group (US), plasmid and ultrasound exposure group (P+US), plasmid+ Lipofectamine group (P+L) were used as controls, respectively. Transfection efficacy was evaluated by observing the red fluorescence in the cells by fluorescent microscopy and flow cytometry(FCM). Ultrasound intensity and exposure time were optimized. Cell apoptosis was investigated using flow cytometry analysis, Hoechst staining, and DNA ladder method. Expression of survivin mRNA was assessed by RT-PCR. Results: Restrictive enzyme digestion and sequencing analysis verified that the recombinant plasmid was successfully constructed. UTMD significantly increased gene transfection efficacy in cultured HeLa cells (P<0.01). Gene transfer was the most prominent at ultrasound intensity of 1.0 W/cm2 and exposure time of 3 min (P<0.01). RT-PCR showed that the expression of survivin mRNA in P+UTMD group was inhibited by (83.33±2.73)%. The differences were significant compared with any other groups (P<0.01). FCM analysis showed that the apoptosis ratio in P+UTMD group was significantly increased as compared with other groups (P<0.01). Hoechst staining and DNA ladder showed that apparent apoptosis and DNA ladder were detected only in P+UTMD and P+L groups. Conclusions:UTMD effectively enhances the transfection efficacy of expression plasmid. It is a novel and effective non-viral gene transfer system and has promising foreground. UTMD mediates RNA interference silenced survivin gene and induces significant cell apoptosis, which provides a new method for tumor research and gene therapy.