Generation of a competing endogenous RNA network and validation of BNIP1 expression in the lung of irradiated mice.

BACKGROUND: Radiation-induced lung injury (RILI) is a serious complication of radiation therapy, and it is mediated by long non-coding RNAs (lncRNAs). STUDY DESIGN AND METHODS: Mouse lung tissues were examined using RNA-Seq and RNA-Seq libraries 72 h after the administration of 6 Gy of X-ray irradiation. The target mRNAs were functionally annotated and the target lncRNA-based miRNAs and target miRNA-based mRNAs were predicted after irradiation to establish the lncRNA-miRNA-mRNA ceRNA axis. RESULTS: The analyses showed that relative to unirradiated controls, 323 mRNAs, 114 miRNAs, and 472 lncRNAs were significantly up-regulated following irradiation, whereas 1907 mRNAs, 77 miRNAs, and 1572 lncRNAs were significantly down-regulated following irradiation. Voltage-gated ion channels, trans-membrane receptor protein tyrosine kinases, and vascular endothelial growth factor have all been associated with dysregulated miRNA-mRNA relationships. KEGG pathway analysis of the dysregulated miRNA-mRNA targets revealed involvement in pathways associated with the hedgehog signaling pathway-fly, ErbB signaling, VEGF signaling, axon guidance, and focal adhesion. KEGG analysis of differentially expressed showed enrichment of mRNAs in primary immunodeficiency, the intestinal immune axis for IgA production, hematopoietic cell lineages, systemic lupus erythematosus, and Th1 and Th2 cell differentiation. Finally, the ceRNA network revealed that BNIP1 was a critical mRNA modulated by the most significant upregulation of lncRNA E230013L22Rik. CONCLUSION: In summary, the lncRNA-miRNA-mRNA ceRNA axis of RILI was constructed following irradiation in a mouse model. RNA dysregulation in the early stage of RILI may lead to severe complications at a later stage, with BNIP1 contributing to radiation-induced cellular apoptosis in RILI.

Evaluation of the lncRNA-miRNA-mRNA ceRNA network in lungs of miR-147 -/- mice.

Background: Previous studies have documented important roles for microRNA-147 (miR-147) in inflammation, radiation-induced injury, cancer, and a range of other diseases. Murine lungs exhibit high levels of miRNA, mRNA, and lncRNA expression. However, very little research to date has focused on the lncRNA-miRNA-mRNA competing endogenous RNA (ceRNA) networks associated with miR-147, and the regulation of lncRNAs and miRNAs in this setting remains poorly understood. Methods: After establishing a miR-147-/- model mouse, samples of lung tissue were harvested for RNA-sequencing, and differentially expressed lncRNAs, miRNAs, and mRNAs were identified. The miRNA targets of these lncRNAs and the identified miRNAs were first overlapped to facilitate the prediction of target mRNAs, with analyses then examining the overlap between these targets and mRNAs that were differentially expressed. Then, these target mRNAs were subjected to pathway enrichment analyses. These results were ultimately used to establish a miR-147-related ceRNA network. Results: Relative to wild-type mice, the lungs of miR-147-/- mice exhibited 91, 43, and 71 significantly upregulated lncRNAs, miRNAs, and mRNAs, respectively, together with 114, 31, and 156 that were significantly downregulated. The lncRNA-miRNA-mRNA network established based on these results led to the identification of Kcnh6 as a differentially expressed hub gene candidate and enabled the identification of a range of regulatory relationships. KEGG pathway enrichment showed that the mRNA targets of differentially expressed lncRNAs and miRNAs in the mice were associated with tumor-related signaling, endometrial cancer, bladder cancer, and ErbB signaling. Conclusion: These results suggest that the identified ceRNA network in miR-147-/- mice shapes tumor-associated signaling activity, with miR-147 potentially regulating various lncRNAs and miRNAs through Kcnh6, ultimately influencing tumorigenesis. Future studies of the lncRNA, miRNA, and mRNA regulatory targets shown to be associated with miR-147 in the present study may ultimately lead to the identification of novel clinically relevant targets through which miR-147 shapes the pathogenesis of cancer and other diseases.

Clinical phenotype and genetic function analysis of a rare family with hereditary leiomyomatosis and renal cell carcinoma complicated with Birt-Hogg-Dubé syndrome.

To date, over 200 families with hereditary leiomyomatosis and renal cell carcinoma (HLRCC) and over 600 families with Birt-Hogg-Dubé (BHD) syndrome have been reported, with low incidence. Here, we describe a patient with suspected rare HLRCC complicated by BHD syndrome. The proband (II1) had characteristic cutaneous leiomyoma-like protrusions on the neck and back, a left renal mass and multiple right renal, liver and bilateral lung cysts. Three family members (I1, II2, II3) had a history of renal cancer and several of the aforementioned clinical features. Two family members (II1, II3) diagnosed with fumarate hydratase (FH)-deficient papillary RCC via pathological biopsy carried two heterozygous variants: FH (NM_000143.3) missense mutation c.1189G>A (p.Gly397Arg) and FLCN (NM_144997.5) frameshift mutation c.1579_1580insA (p.Arg527Glnfs*75). No family member carrying a single variant had renal tumours. In HEK293T cells transfected with mutant vectors, mRNA and protein expression after FLCN p.Arg527Glnfs*75 and FH p.Gly397Arg mutations were significantly lower than those in wild-type (WT) cells. Cell immunofluorescence showed altered protein localisation and reduced protein expression after FLCN p.Arg527Glnfs*75 mutation. The FH WT was uniformly distributed in the cytoplasm, whereas FH protein expression was reduced after the p.Gly397Arg mutation and scattered sporadically with altered cell localisation. Patients with two variants may have a significantly increased penetrance of RCC.

PARP-1: a critical regulator in radioprotection and radiotherapy-mechanisms, challenges, and therapeutic opportunities.

Background: Since its discovery, poly (ADP-ribose) polymerase 1 (PARP-1) has been extensively studied due to its regulatory role in numerous biologically crucial pathways. PARP inhibitors have opened new therapeutic avenues for cancer patients and have gained approval as standalone treatments for certain types of cancer. With continued advancements in the research of PARP inhibitors, we can fully realize their potential as therapeutic targets for various diseases. Purpose: To assess the current understanding of PARP-1 mechanisms in radioprotection and radiotherapy based on the literature. Methods: We searched the PubMed database and summarized information on PARP inhibitors, the interaction of PARP-1 with DNA, and the relationships between PARP-1 and p53/ROS, NF-κB/DNA-PK, and caspase3/AIF, respectively. Results: The enzyme PARP-1 plays a crucial role in repairing DNA damage and modifying proteins. Cells exposed to radiation can experience DNA damage, such as single-, intra-, or inter-strand damage. This damage, associated with replication fork stagnation, triggers DNA repair mechanisms, including those involving PARP-1. The activity of PARP-1 increases 500-fold on DNA binding. Studies on PARP-1-knockdown mice have shown that the protein regulates the response to radiation. A lack of PARP-1 also increases the organism's sensitivity to radiation injury. PARP-1 has been found positively or negatively regulate the expression of specific genes through its modulation of key transcription factors and other molecules, including NF-κB, p53, Caspase 3, reactive oxygen species (ROS), and apoptosis-inducing factor (AIF). Conclusion: This review provides a comprehensive analysis of the physiological and pathological roles of PARP-1 and examines the impact of PARP-1 inhibitors under conditions of ionizing radiation exposure. The review also emphasizes the challenges and opportunities for developing PARP-1 inhibitors to improve the clinical outcomes of ionizing radiation damage.

Clinical and imaging features of six Han patients with SAPHO syndrome.

BACKGROUND: Synovitis, acne, pustulosis, hyperostosis, and osteitis (SAPHO) syndrome is a rare autoimmune disease characterized by skin or osteoarticular damage. SAPHO syndrome is often misdiagnosed or missed diagnosis due to lack of overall understanding of the disease by clinicians. PURPOSE: To analyze the clinical symptoms and imaging features of six Han patients with SAPHO syndrome in order to provide reference for doctors to diagnose SAPHO syndrome. MATERIAL AND METHODS: This study retrospectively analyzed the clinical data of six Han patients with SAPHO syndrome. RESULTS: All six Han patients with SAPHO syndrome had severe acne or pustulosis of the hands and feet, and all of them had osteoarticular damage, including five cases involving the sternoclavicular joint. Some patients showed a specific and typical "bull's head" sign on 99mTc-labeled methylene diphosphonate bone imaging. Among the six patients recruited, there was one thoracic vertebra, one cervical vertebra, one sacroiliac joint, and one peripheral joint involvement. Two patients had limited activity due to severe osteoarticular damage. CONCLUSION: Due to the atypical clinical symptoms of SAPHO syndrome, most patients will experience a tortuous and long diagnostic process, while a correct understanding and timely intervention of SAPHO syndrome are essential to improve the prognosis of patients.

Case report and analysis: Behçet's disease with lower extremity vein thrombosis and pseudoaneurysm.

Background: Behçet's disease (BD) is a unique autoimmune chronic systemic vasculitis that affects veins and arteries of all sizes. BD can lead to recurrent vascular events, especially venous thrombosis, with an incidence rate of 40%, or pseudoaneurysms formed under long-term inflammatory reaction or iatrogenic stimulation. BD-related risk factors promote endothelial dysfunction, platelet activation and overactivation of tissue factors leading to mural inflammatory thrombi. Thrombosis may be the first clinical manifestation of BD. Case presentation: A 32-year-old man complaining of progressive swelling and pain in the right lower extremity for 30 days was initially diagnosed with "venous thrombosis of the right lower extremity," using color Doppler ultrasonography. Patient underwent inferior vena cava filter placement combined with deep vein angioplasty of the right lower extremity and catheter-directed urokinase thrombolysis. Postoperative oral anticoagulant therapy was administered. However, the patient was readmitted 20 days later for pulsatile pain in the right groin. Prior medical history included 4 years of repeated oral and perineal ulcers, and 2 months of blurred vision. Abdominal computed tomography angiography (CTA) revealed rupture of the right common iliac artery (CIA) and left internal iliac artery (IIA), complicated by a pseudoaneurysm. Based on the clinical manifestations and other auxiliary examination results, the patient was re-diagnosed with "BD combined with deep venous thrombosis of the right lower extremity and an iliac artery pseudoaneurysm." Stent implantation was performed for iliac artery pseudoaneurysm after symptoms were controlled with timely immunosuppressive therapy. After endovascular treatment, the patient underwent continued immunosuppressive therapy and dynamic reexaminations of abdominal CTA, which revealed that a small amount of contrast agent at the stent in the right CIA continued to flow into the cavity of the pseudoaneurysm; in addition, the size of the pseudoaneurysm was gradually increasing. Therefore, the patient underwent a second stent implantation for iliac artery pseudoaneurysm, and the condition improved further. Conclusion: The importance of early diagnosis of BD should be recognized, and the choice of interventional and surgical procedures should be carefully evaluated, as this may trigger further damage to vascular access in BD patients with aneurysm.

Association of menopausal status with COVID-19 outcomes: a propensity score matching analysis.

BACKGROUND: Despite the growing number of studies on the coronavirus disease-19 (COVID-19), little is known about the association of menopausal status with COVID-19 outcomes. MATERIALS AND METHODS: In this retrospective study, we included 336 COVID-19 inpatients between February 15, 2020 and April 30, 2020 at the Taikang Tongji Hospital (Wuhan), China. Electronic medical records including patient demographics, laboratory results, and chest computed tomography (CT) images were reviewed. RESULTS: In total, 300 patients with complete clinical outcomes were included for analysis. The mean age was 65.3 years, and most patients were women (n = 167, 55.7%). Over 50% of patients presented with comorbidities, with hypertension (63.5%) being the most common comorbidity. After propensity score matching, results showed that men had significantly higher odds than premenopausal women for developing severe disease type (23.7% vs. 0%, OR 17.12, 95% CI 1.00-293.60; p = 0.003) and bilateral lung infiltration (86.1% vs. 64.7%, OR 3.39, 95% CI 1.08-10.64; p = 0.04), but not for mortality (2.0% vs. 0%, OR 0.88, 95% CI 0.04-19.12, p = 1.00). However, non-significant difference was observed among men and postmenopausal women in the percentage of severe disease type (32.7% vs. 41.7%, OR 0.68, 95% CI 0.37-1.24, p = 0.21), bilateral lung infiltration (86.1% vs. 91.7%, OR 0.56, 95% CI 0.22-1.47, p = 0.24), and mortality (2.0% vs. 6.0%, OR 0.32, 95% CI 0.06-1.69, p = 0.25). CONCLUSIONS: Men had higher disease severity than premenopausal women, while the differences disappeared between postmenopausal women and men. These findings support aggressive treatment for the poor prognosis of postmenopausal women in clinical practice.

Genetic diagnosis history and osteoarticular phenotype of a non-transfusion secondary hemochromatosis.

BACKGROUND: It is not easy to identify the cause of various iron overload diseases because the phenotypes overlap. Therefore, it is important to perform genetic testing to determine the genetic background of patients. AIM: To investigate the genetic background of a patient with hemochromatosis complicated by psoriasis on both lower extremities. METHODS: Ten years ago, a 61-year-old male presented with iron overload, jaundice, hemolytic anemia and microcytic hypochromic anemia. Computed tomography of the left knee joint showed enlargement of the tibial medullary cavity and thinned bone cortices. Magnetic resonance imaging showed hepatic hemochromatosis, extensive abnormal signals from bone marrow cavities and nodular lesions in the lateral medullary cavity of the upper left lateral tibia. Single photon emission computed tomography showed radial dots of abnormal concentration in the upper end of the left tibia and radial symmetry of abnormal concentrations in joints of the extremities. The patient showed several hot spot mutations of the HFE and G6PD genes detected by next-generation sequencing, but no responsible gene mutation was found. The thalassemia gene was detected by gap-PCR. RESULTS: The patient was found to carry the -α4.2 and --SEA deletion mutations of the globin gene. These two mutations are common causes of Southeast Asian α-thalassemia, but rarely cause severe widespread non-transfusion secondary hemochromatosis osteoarthropathy. The simultaneous presence of an auxiliary superposition effect of a rare missense mutation of the PIEZO1 gene (NM_001142864, c.C4748T, p.A1583V) was considered. Moreover, several rare mutations of the IFIH1, KRT8, POFUT1, FLG, KRT2, and TGM5 genes may be involved in the pathogenesis of psoriasis. CONCLUSION: The selection of genetic detection methods for hemochromatosis still needs to be based on an in-depth study of the clinical manifestations of the disease.

Complicated Posterior Cervical Skin and Soft Tissue Infections at a Single Referral Center.

BACKGROUND: Although the incidence and mortality of complicated skin and soft tissue infections have decreased, this infection is still relatively frequent and can be associated with lethal complications. In this study, the authors present our clinical experience of patients with complicated posterior cervical skin and soft tissue infections (CPCSSTIs) diagnosed and treated in a reconstructive unit in northeastern China. METHODS: A retrospective chart review of patients diagnosed with CPCSSTIs from January 2009 to December 2018 was performed. To make the results objective and convincing, a data analysis was performed relating to demographic characteristics, clinical presentation, predisposing factor, bacterial culture, laboratory and radiographic evaluations, diagnostic clues, management, and complications as well as the clinical course and outcome. RESULTS: During the ten-year period, there were 174 consecutive patients admitted to our reconstructive center with final diagnosis of CPCSSTIs included. All the patients were adults, and the majority were male (67.2%). The patient's mean age was 51.3 years (range, 15-88 years). There were 114 patients (65.5%) that had associated systemic diseases, with diabetes mellitus (40.2%) as the most common predisposing factor. Common presented clinical symptoms were pain (90.8%), swelling (85.1%), and erythema (77%) of the neck. Surgical treatment was performed in all the patients, and most of them (83.9%) received the first surgery within 24 h. The most commonly isolated pathogen was Staphylococcus aureus (30%). Vancomycin (21.3%) was the most commonly used antibiotics, followed by cefepime (18.4%). All the patients survived and were discharged with a mean duration of hospitalization of 28.7 days. Those patients with predisposing factors (31.4 ± 12.35 days) or complications (41.0 ± 12.5 days) tended to have a longer hospital stay. The mean total costs of admission per patient were 47 644 RMB. CONCLUSION: This study highlights the high cost burden of CPCSSTI patients. Those patients with predisposing factors or complications tended to have a longer hospital stay.

Characterisation of the complete chloroplast genome of an endemic herb plant species, Rheum palmatum (Polygonaceae) in China.

Rheum palmatum has a long history in medicine, which is one of the main export medicinal herb in China. The complete chloroplast genome of R. palmatum was assembled and reported in this study. The R. palmatum chloroplast genome was 161,541 bp in length as the circular and consisted a large single-copy (LSC) region of 86,519 bp, a small single-copy (SSC) region of 13,112 bp and a pair of inverted-repeat (IR) regions of 30,955 bp. The nucleotide composition was asymmetric 31.2% A (Adenine), 31.5% T (Thymine), 19.0% C (Cytosine), and 18.3% G (Guanine) with an overall G + C content of 37.3%. It encoded 131 genes, including 86 protein-coding genes (76 PCG species), 37 transfer RNA genes (26 tRNAs species), and eight ribosomal RNA genes (four rRNAs species). The Phylogenetic relationships used neighbour-joining (NJ) method and the result showed that R. palmatum and Rheum officinale are phylogenetically related to each other in the family Polygonaceae. This study will be very important for Chinese medicinal herb research value and clinical drug development for future in China.

Lower energy radial shock wave therapy improves characteristics of hypertrophic scar in a rabbit ear model.

The aim of the present study was to investigate the effects of radial extracorporeal shock wave therapy (rESWT) on scar characteristics and transforming growth factor (TGF)-ß1/Smad signaling in order to explore a potential modality for the treatment of hypertrophic scars (HS). The HS model was generated in rabbit ears, then rabbits were randomly divided into 3 groups: Lower (L)-ESWT [treated with rESWT with lower energy flux density (EFD) of 0.1 mJ/mm2], higher (H)-ESWT (treated with a higher EFD of 0.18 mJ/mm2) and the sham ESWT group (S-ESWT; no ESWT treatment). Scar characteristics (wrinkles, texture, diameter, area, volume of elevation, hemoglobin and melanin) were assessed using the Antera 3D® system. The protein and mRNA expression of TGF-ß1, Smad2, Smad3 and Smad7 was assessed by enzyme-linked immunosorbent assay and reverse transcription-quantitative polymerase chain reaction, respectively. The Antera 3D® results indicated that wrinkles and hemoglobin of the HS were significantly improved in both of the rESWT groups when compared with the S-ESWT group. However, these changes appeared much earlier in the L-ESWT group than the H-ESWT. Scar texture was also improved in the L-ESWT group. However, rESWT did not influence HS diameter, area, volume of elevation or melanin levels. rESWT had no effect on TGF-ß1 or Smad7 expression in either of rESWT groups. Although no difference was observed in Smad2 mRNA expression in the L-ESWT group, the Smad3 mRNA and protein expression significantly decreased when compared with the H-ESWT and S-ESWT groups. By contrast, Smad2 and Smad3 mRNA expression were upregulated in the H-ESWT group. These results demonstrated that rESWT with 0.1 mJ/mm2 EFD improved some characteristics of the HS tissue. Downregulation of Smad3 expression may underlie this inhibitory effect. Inhibition of the TGF-ß1/Smad signal transduction pathway may be a potential therapeutic target for the management of HS.

Couple-kissing flaps for successful repair of severe sacral pressure ulcers in frail elderly patients.

BACKGROUND: Surgical repair of severe pressure ulcers (PUs) in elderly patients remains a challenge for clinicians due to the complicated comorbidities and the special physical characteristics of elderly patients. The objective of this study was to evaluate the application of couple-kissing flaps (CKF) in the reconstruction of sacral PUs in these patients. METHODS: Elderly patients (over 70 years) with stage 3 or stage 4 PUs who underwent CKF immediately after radical debridement between July 2012 and December 2015 were enrolled in this retrospective study. Patients' demographics were extracted from the medical records. RESULTS: A total of 12 patients were involved in this study. The average age of the patients was 76.83 years (ranged from 71 to 92 years). The donor site was closed primarily in all cases. All the flaps healed uneventfully without complications. Follow-up observations were conducted for an average of 13.6 months (ranged from 9 months to 2 years). Cosmetic results were satisfactory, with no surgical site breakdown or recurrence of PU in any of the cases. Three representative cases are presented. CONCLUSIONS: The CKF is a reliable and satisfactory option for the reconstruction of severe sacral PUs defects in elderly patients. CKF is associated with an relatively low rate of complications and recurrence.

Necrotizing soft tissue infection: clinical characteristics and outcomes at a reconstructive center in Jilin Province.

BACKGROUND: To investigate the clinical characteristics and treatment outcomes in necrotizing fasciitis (NF) patients in a reconstructive unit in northeastern China. METHODS: Medical records of patients diagnosed with and treated for NF in the extremities from November 2013 to December 2016 were retrospectively reviewed. Demographic data, clinical presentation, duration of signs and symptoms, location of infection, predisposing factors, causative microbiological organisms, laboratory risk indicator for necrotizing fasciitis (LRINEC) score, number of surgical debridements, length of hospital stay, treatments, and outcomes were recorded. RESULTS: A total of 39 consecutive patients were treated for severe NF (32 male and 7 female). Diabetes mellitus and blunt trauma were the most common risk factors (13 and 9 cases, respectively). The positive predictive value of the LRINEC score in NF diagnosis was 46.2%. Mean duration of signs and symptoms was 4.6 days. Staphylococcus aureus was the most commonly isolated bacteria (20 cases). All patients underwent their first debridement within 12 h of presentation (mean, 4.6 h). Mean number of surgical treatments was 2.8 (range, 2-5) per patient, including debridements. All patients survived, and mean length of hospital stay was 30.81 (range, 21-43) days. Three patients underwent limb amputation. CONCLUSIONS: In our clinical experience, early detection and aggressive debridement are the cornerstones of NF treatment. Antibiotic therapy and intensive care support is essential in severe cases of NF. Anaerobic tissue culture and frozen section biopsy could be adopted as routine tests for diagnosis and decision-making in NF. These findings should inform clinical decisions about the treatment of individual patients with NF.

Transcriptome analysis of the mammary gland from GH transgenic goats during involution.

Mammary glands are organs for milk production in female mammals. Growth hormone (GH) is known to affect the growth and development of the mammary gland, as well as to increase milk production in dairy goats. This study performed a comprehensive expression profiling of genes expressed in the mammary gland of early involution GH transgenic (n=4) and non-transgenic goats (n=4) by RNA sequencing. RNA was extracted from mammary gland tissues collected at day 3 of involution. Gene expression analysis was conducted by Illumina RNA sequencing and sequence reads were assembled and analyzed using TopHat. FPKM (fragments per kilobase of exon per million) values were analyzed for differentially expressed genes using the Cufflinks package. Gene ontology analysis of differentially expressed genes was categorized using agriGO, while KEGG pathway analysis was performed with the online KEGG automatic annotation server. Our results revealed that 75% of NCBI goat annotated genes were expressed during early involution. A total of 18,323 genes were expressed during early involution in GH transgenic goats, compared with 18,196 expressed genes during early involution of non-transgenic goats. In these expressed genes, the majority (17,589) were ubiquitously expressed in GH transgenic and non-transgenic goats. However, there were 745 differentially expressed genes, 421 of which were upregulated and 324 were downregulated in GH transgenic goats. GO and KEGG pathway analysis showed that these genes were involved in mammary gland physiology, including cell adhesion molecules, ECM-receptor interaction, Jak-STAT signaling pathway, and fat metabolism. Our results demonstrated that the GH receptor was strongly affected in GH transgenic goats, which may activate the IGF-1/Stat3 signaling pathway. Overall, our study provided a global view of the transcriptome during involution of GH transgenic and non-transgenic goats, which increases our understanding of the biology of involution in the goat.

Enhancement of transfection efficiency with NLS and SPB-NLS.

Low positive cell screening efficiency severely hinders the development of transgenic animals. The major rate-limiting step of positive cell screening is DNA entering the nucleus, particularly for large DNA molecules. To enhance the transport of large DNA molecules into the nucleus, particularly for the production of transgenic animals, nuclear localization sequence (NLS) peptides and the peptide derivative succinimidyl-[4-(psoralen-8-yloxy)]-butyrate (SPB)-NLS were synthesized to mediate transfection in vitro. To investigate the function of NLS and SPB-NLS in vitro, the expression levels of growth hormone (GH) mRNA and green fluorescent protein (GFP) protein were analyzed following transfection mediated by NLS and SPB-NLS. The results demonstrated that the expression of GH mRNA was significantly higher in the NLS (increased by 69%) and SPB-NLS (330%) groups than that in the liposome/pGN group. Similarly, GFP expression was found to be higher in the SPB-NLS group than that in the liposome group, while the expression in the NLS group was lower than that in the liposome group. Further analysis demonstrated that SPB-NLS enhanced the expression of insulin-like growth factor 1 in hard-to-transfect goat mammary epithelia cells. The results of the microscopy analysis revealed that transfected DNA entered the nucleus via the nuclear pores, facilitated by NLS. Analysis of the cell cycle demonstrated that the cytotoxic effects of NLS and SPB-NLS were low. In conclusion, the results of the present study demonstrate that SPB-NLS acts as a transfection-enhancing agent and may be used both to enhance nuclear delivery and for the development of genetically modified animals.

Enhancement of gene transfer efficiency in the Bcap-37 cell line by dimethyl sulphoxide and menthol.

Simple and efficient gene transfer into the nucleus would facilitate non-viral gene delivery. One promising method of non-viral gene delivery is to apply penetration enhancers. Chemicals, such as dimethyl sulfoxide (DMSO) and menthol, may have promise as non-toxic vehicles in improving gene transfer efficiency. In this study, the cytotoxic effects of DMSO and menthol were evaluated using MTT assays. Gene delivery efficiency in a human breast cancer cell line (Bcap-37) was investigated by quantitative PCR, fluorescence microscopy and flow cytometry. Non-toxic concentrations of DMSO (2%) and menthol (12.5 µM) enhanced the efficiency of liposome-mediated gene delivery in Bcap-37 cells. Quantitative PCR results showed that growth hormone (GH) mRNA expression in the post-menthol and pre-DMSO treatment groups was 10-fold higher compared to that in the liposome group, while in the pre-menthol and post-DMSO treatment groups, a 30-fold increase in GH mRNA expression was observed. Both DMSO and menthol treatments increased green fluorescent protein (GFP) expression efficiency as shown by fluorescence microscopy experiments. Compared to the liposome group, the number of positive cells in the pre-menthol and post-DMSO treatment groups was significantly increased by 15%. Furthermore, cell cycle analysis demonstrated that there were significant differences among the DMSO-treated group, the menthol-treated group and the normal group, which implied different effects of DMSO and menthol treatments. In conclusion, both non-toxic and harmless DMSO (2%) and menthol (12.5 µM) treatments improve gene transfer efficiency, while post-DMSO treatment may be the most effective protocol in increasing transgene expression efficiency.

[Design and experiment of micro biochemical detector based on micro spectrometer].

According to the requirements of rapid detection of important life parameters for the sick and wounded, a new micro bio-chemical detection configuration was proposed utilizing continuous spectroscopy analysis, which was founded on MOEMS and embedded technology. The configuration was developed as so much research work was carried out on the detecting objects and methods. Important parameters such as stray light, absorbance linearity, absorbance ratability, stability and temperature accuracy of the instrument were tested, which are all in good agreement with the design requirements. Clinic tests show that it can detect multiple life parameters quickly (Na+, GLU, Hb eg.).

Diversity of tight junctions (TJs) between gastrointestinal epithelial cells and their function in maintaining the mucosal barrier.

The gastrointestinal epithelium, which is covered by a single layer of epithelial cells, including enterocytes, intraepithelial lymphocytes, goblet cells, microfold cells, and dendritic cells, serves as a protective barrier separating luminal contents from the underlying tissue compartments. The epithelium plays an important role in the first line of host defense against a variety of pathogens, as well as maintaining the homeostasis in gastrointestinal tract. All these epithelial cells express junction complex proteins and form cell junctions such as adherens and TJs, although the TJs have small differences among different epithelial cells. The TJs, located most apically on the lateral membrane, are required for the proper formation of epithelial cell polarity as well as sustaining of the mucosal barrier. Furthermore, TJs are the key cell junctions modulating the paracellular pathway. Understanding the diversity of the TJs between intestinal epithelial cells and their different roles in defending pathogens' invasion and modifying the paracellular pathway are attractive to exploration.

Use of green fluorescent protein to monitor Lactobacillus in the gastro-intestinal tract of chicken.

Lactobacilli, like other gut commensal bacteria, are well known for their use in industrial food fermentations and for their probiotic properties. However, little is known about the interaction of these microorganisms with the gastro-intestinal epithelia when administered in vivo. To specifically monitor the passage of lactobacilli after oral administration, the gfp gene was cloned downstream from the constitutive l-lactate dehydrogenase promoter (pldhL) in the experiment. The recombinant expression vector pLEM415::gfp was electroporated into different lactobacilli isolated from chicken. Green fluorescent protein (GFP) was expressed successfully in Lactobacillus delbrueckii ssp. lactis D17 (D17-GFP) and Lactobacillus fructosus C2 (C2-GFP). Moreover, oral administration of D17-GFP in chickens allowed us to trace it in the gastro-intestinal tract. Six hours after ingestion, D17-GFP was detectable in all luminal contents (stomach, jejunum, ileum and caecum). At 42 h post-administration the microorganism was present throughout the intestine with maximum concentrations about 10(5.5) in all intestinal sections. No fluorescent lactobacilli were detected in the spleen or liver of chickens at any time. Using fluorescence microscopy, it became apparent that the D17-GFP were mainly embedded in the mucus, localized close to the epithelial surface of the intestine and scattered in the intestine lamina propria.