Cystic fibrosis (CF) is a rare autosomal recessive disease with only one pathogenic gene cystic fibrosis transmembrane conductance regulator (CFTR). To identify the potential pathogenic mutations in a Chinese patient with CF, we conducted Sanger sequencing on the genomic DNA of the patient and his parents and detected all 27 coding exons of CFTR and their flanking intronic regions. The patient is a compound heterozygote of c.2909G > A, p.Gly970Asp in exon 18 and c.1210-3C > G in cis with a poly-T of 5T (T5) sequence, 3 bp upstream in intron 9. The splicing effect of c.1210-3C > G was verified via minigene assay in vitro, indicating that wild-type plasmid containing c.1210-3C together with T7 sequence produced a normal transcript and partial exon 10-skipping-transcript, whereas mutant plasmid containing c.1210-3G in cis with T5 sequence caused almost all mRNA to skip exon 10. Overall, c.1210-3C > G, the newly identified pathogenic mutation in our patient, in combination with T5 sequence in cis, affects the CFTR gene splicing and produces nearly no normal transcript in vitro. Moreover, this patient carries a p.Gly970Asp mutation, thus confirming the high-frequency of this mutation in Chinese patients with CF.
China ; Cystic Fibrosis/genetics* ; Cystic Fibrosis Transmembrane Conductance Regulator/genetics* ; Humans ; Mutation ; Poly T ; RNA, Messenger/genetics*

BACKGROUND: Wound healing mechanisms is believed to have effects similar to wound healing disorders in diabetic patients, including abnormal inflammatory cells, angiogenesis disorders, and reduced collagen synthesis. Therefore, reestablishment of structural and promoted angiogenesis could be beneficial to promote wound healing process. OBJECTIVE: Therefore, we investigated whether the polydeoxyribonucleotide (PDRN) that was self-production in Korea, could be useful as an intradermal injection for promoting wound healing. Also, we validate for wound healing effect of PDRN using healing-impaired (db/db) mice. METHODS: In this study, we confirmed the effects of PDRN by creating wound models in in vitro and in vivo model. Using an in vitro wound healing assay, we observed that PDRN stimulated closure of wounded monolayers of human fibroblast cells. PDRN (8.25 mg/ml) or phosphate-buffered saline (0.9% NaCl) was injected once daily into the dermis adjacent to the wound for 12 days after skin injury. RESULTS: Time course observations revealed that mice treated with PDRN showed accelerated wound closure and epidermal and dermal regeneration, enhanced angiogenesis. The wound area and depth decreased at 3, 6, 9, and 12 days after skin injury. Histological evaluation showed an increase of vascular endothelial growth factor, CD31, and collagen fibers in the PDRN group compared with the control group, indicating that PDRN was effective in the treatment of delayed wound healing caused by diabetes. CONCLUSION: This study suggests that our PDRN has a wound healing effect in transgenic animal models with cells and diabetes through angiogenesis.
Animals ; Animals ; Animals, Genetically Modified ; Collagen ; Dermis ; Fibroblasts ; Humans ; In Vitro Techniques ; Injections, Intradermal ; Korea ; Mice ; Models, Animal ; Polydeoxyribonucleotides ; Regeneration ; Skin ; Vascular Endothelial Growth Factor A ; Wound Healing ; Wounds and Injuries

OBJECTIVE: To assess the clinical efficacy and safety of intra-articular injection of hyaluronic acid (HA) combined with polydeoxyribonucleotide (PDRN) in patients with knee osteoarthritis in comparison with that of HA alone. METHODS: The current single-center, prospective, randomized, double-blind, controlled study was conducted in 36 patients with knee osteoarthritis at our medical institution. All the eligible patients (n=30) were equally assigned to two treatment arms (trial group ‘HA+PDRN’ and control group ‘HA’). For efficacy assessment, the patients were evaluated for the visual analogue scale (VAS) scores, the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and the Knee Society Scores (KSS), all of which served as efficacy outcome measures. We monitored time-dependent changes in efficacy outcome measures at baseline and 1, 3 and 6 months. Subsequently, we compared differences in changes in efficacy outcome measures at 6 months from baseline between the two groups. Moreover, we assessed the safety based on the treatment-emergent adverse events (TEAEs), adverse drug reactions (ADRs) and any other complications serving as safety outcome measures. RESULTS: There were significant differences in changes in the VAS scores, the WOMAC scores in all domains, except ‘Stiffness’, the total WOMAC scores, and the KSS scores in all the domains at 6 months from baseline between the two groups (p<0.05). In our series, there were no TEAEs, ADRs, and any other complications. CONCLUSION: Intra-articular injections of HA combined with PDRN can also be considered in the treatment of knee osteoarthritis. However, further large-scale and multi-center studies are required to demonstrate the potential of the proposed combination.
Arm ; Drug-Related Side Effects and Adverse Reactions ; Humans ; Hyaluronic Acid ; Injections, Intra-Articular ; Knee ; Ontario ; Osteoarthritis ; Osteoarthritis, Knee ; Outcome Assessment (Health Care) ; Polydeoxyribonucleotides ; Prospective Studies ; Treatment Outcome

Polydeoxyribonucleotide (PDRN) is safe and effective in wound healing, cellular growth, synthesis of extracellular matrix protein, and inflammation reduction via activation of adenosine A2 receptors. We report a 28-year-old male patient treated with PDRN injections for chronic non-healing wound refractory to negative pressure wound therapy, skin graft, or growth factors. Three injections of PDRN were administered at the wound site into the anterior and medial sides of the left stump on the 1st, 4th, and 9th days of hospitalization. The PDRN ameliorated wound healing by enhancing cell growth, tissue repair, and angiogenesis. PDRN application represents a potential treatment for non-healing wounds obviating the need for additional therapies, and hospitalization, as well as improve patient’s activities of daily living.
Activities of Daily Living ; Adult ; Amputees* ; Extracellular Matrix ; Hospitalization ; Humans ; Inflammation ; Intercellular Signaling Peptides and Proteins ; Male ; Negative-Pressure Wound Therapy ; Polydeoxyribonucleotides ; Receptors, Adenosine A2 ; Skin ; Transplants ; Wound Healing ; Wounds and Injuries*

Interleukin (IL)-17 plays an important role in rhinitis and the level thereof correlates with the severity of disease. However, no mouse model for IL-17-dominant rhinitis has yet been developed. Our objective was to establish a mouse model of IL-17-dominant rhinitis via intranasal application of polyinosinic-polycytidylic acid (abbreviated as Poly(I:C)). Mice were divided into 6 groups (n=8 for each group); 1) 1 negative control group, 2) 1 positive control group (OVA/alum model), 3) 2 Poly(I:C) groups (10 or 100 µg), and 4) 2 OVA/Poly(I:C) groups (10 or 100 µg). The positive control group was treated with the conventional OVA/alum protocol. In the Poly(I:C) and OVA/Poly(I:C) groups, phosphate-buffered saline or an OVA solution plus Poly(I:C) were administered. The OVA/Poly(I:C) groups exhibited significantly greater neutrophil infiltration and increased IL-17/interferon γ expression compared with the other groups. However, the levels of total immunoglobulin E (IgE), OVA-specific IgE, eosinophil infiltration, IL-4, IL-5, IL-6, and IL-10 were significantly lower in the OVA/Poly(I:C) groups than in mice subjected to conventional Th2-dominant OVA/alum treatment (the positive control group). IL-17 and neutrophil measurement, chemokine (C-X-C motif) ligand 1 immunohistochemistry, and confocal microscopy revealed increased numbers of IL-17-secreting cells in the nasal mucosa of the OVA/Poly(I:C) groups, which included natural killer cells, CD4 T cells, and neutrophils. In conclusion, we developed a mouse model of IL-17-dominant rhinitis using OVA together with Poly(I:C). This model will be useful in research on neutrophil- or IL-17-dominant rhinitis.
Animals ; Chemokine CXCL1 ; Eosinophils ; Immunoglobulin E ; Immunoglobulins ; Immunohistochemistry ; Interleukin-10 ; Interleukin-17 ; Interleukin-4 ; Interleukin-5 ; Interleukin-6 ; Interleukins ; Killer Cells, Natural ; Mice* ; Microscopy, Confocal ; Nasal Mucosa ; Neutrophil Infiltration ; Neutrophils ; Ovum ; Poly I-C* ; Rhinitis* ; T-Lymphocytes

BACKGROUND: Polydeoxyribonucleotide (PDRN) is known to have anti-inflammatory and angiogenic effects and to accelerate wound healing. The aim of this study was to investigate whether PDRN could improve peripheral tissue oxygenation and angiogenesis in diabetic foot ulcers. METHODS: This was a prospective randomized controlled clinical trial. Twenty patients with a non-healing diabetic foot ulcer were randomly distributed into a control group (n=10) and a PDRN group (n=10). Initial surgical debridement and secondary surgical procedures such as a split-thickness skin graft, primary closure, or local flap were performed. Between the initial surgical debridement and secondary surgical procedures, 0.9% normal saline (3 mL) or PDRN was injected for 2 weeks by the intramuscular (1 ampule, 3 mL, 5.625 mg, 5 days per week) and perilesional routes (1 ampule, 3 mL, 5.625 mg, 2 days per week). Transcutaneous oxygen tension (TcPO2) was evaluated using the Periflux System 5000 with TcPO2/CO2 unit 5040 before the injections and on days 1, 3, 7, 14, and 28 after the start of the injections. A pathologic review (hematoxylin and eosin stain) of the debrided specimens was conducted by a pathologist, and vessel density (average number of vessels per visual field) was calculated. RESULTS: Compared with the control group, the PDRN-treated group showed improvements in peripheral tissue oxygenation on day 7 (P < 0.01), day 14 (P < 0.001), and day 28 (P < 0.001). The pathologic review of the specimens from the PDRN group showed increased angiogenesis and improved inflammation compared with the control group. No statistically significant difference was found between the control group and the PDRN group in terms of vessel density (P=0.094). Complete healing was achieved in every patient. CONCLUSIONS: In this study, PDRN improved peripheral tissue oxygenation. Moreover, PDRN is thought to be effective in improving inflammation and angiogenesis in diabetic foot ulcers.
Angiogenesis Modulating Agents ; Blood Gas Monitoring, Transcutaneous ; Debridement ; Diabetic Foot* ; Eosine Yellowish-(YS) ; Foot Ulcer ; Humans ; Inflammation ; Oxygen* ; Polydeoxyribonucleotides ; Prospective Studies ; Skin ; Transplants ; Ulcer* ; Wound Healing

BACKGROUND: Polydeoxyribonucleotide (PDRN) is known to have anti-inflammatory and angiogenic effects and to accelerate wound healing. The aim of this study was to investigate whether PDRN could improve peripheral tissue oxygenation and angiogenesis in diabetic foot ulcers. METHODS: This was a prospective randomized controlled clinical trial. Twenty patients with a non-healing diabetic foot ulcer were randomly distributed into a control group (n=10) and a PDRN group (n=10). Initial surgical debridement and secondary surgical procedures such as a split-thickness skin graft, primary closure, or local flap were performed. Between the initial surgical debridement and secondary surgical procedures, 0.9% normal saline (3 mL) or PDRN was injected for 2 weeks by the intramuscular (1 ampule, 3 mL, 5.625 mg, 5 days per week) and perilesional routes (1 ampule, 3 mL, 5.625 mg, 2 days per week). Transcutaneous oxygen tension (TcPO2) was evaluated using the Periflux System 5000 with TcPO2/CO2 unit 5040 before the injections and on days 1, 3, 7, 14, and 28 after the start of the injections. A pathologic review (hematoxylin and eosin stain) of the debrided specimens was conducted by a pathologist, and vessel density (average number of vessels per visual field) was calculated. RESULTS: Compared with the control group, the PDRN-treated group showed improvements in peripheral tissue oxygenation on day 7 (P < 0.01), day 14 (P < 0.001), and day 28 (P < 0.001). The pathologic review of the specimens from the PDRN group showed increased angiogenesis and improved inflammation compared with the control group. No statistically significant difference was found between the control group and the PDRN group in terms of vessel density (P=0.094). Complete healing was achieved in every patient. CONCLUSIONS: In this study, PDRN improved peripheral tissue oxygenation. Moreover, PDRN is thought to be effective in improving inflammation and angiogenesis in diabetic foot ulcers.
Angiogenesis Modulating Agents ; Blood Gas Monitoring, Transcutaneous ; Debridement ; Diabetic Foot* ; Eosine Yellowish-(YS) ; Foot Ulcer ; Humans ; Inflammation ; Oxygen* ; Polydeoxyribonucleotides ; Prospective Studies ; Skin ; Transplants ; Ulcer* ; Wound Healing

Complex regional pain syndrome (CRPS) type II is a syndrome that develops after nerve injury. Symptoms may be severe, and vary depending on the degree of sympathetic nerve involvement. As yet, there is no satisfactory treatment. We report the case of a female patient who had an L5 left transverse process fracture and an S2 body fracture, who developed symptoms of CRPS type II in her left lower leg that were aggravated during ambulation in spite of absolute bed rest for one month after the trauma. Several treatments, including bed rest, medication, and numerous nerve blocks were attempted, but the pain persisted. We finally tried injection of polydeoxyribonucleotide (PDRN) solution at the left L5 transverse process fracture site because we knew of the anti-inflammatory effect of PDRN. One day after this treatment, her symptoms had almost disappeared and three days later, she was discharged. We will also further discuss the possibility of using PDRN solution for the treatment of CRPS.
Bed Rest ; Causalgia* ; Female ; Humans ; Leg ; Nerve Block ; Polydeoxyribonucleotides ; Walking

OBJECTIVETo explore the effect of a new emodin derivative E11 on proliferation and apoptosis of T lymphocytic leukemia cell line Molt-4 and its possible mechanisms.

METHODSMTT method was used to plot cell growth curve. Colony culture assay was performed for studying the effect of emodin derivative E11 on colony-formation of Molt-4. The fluorescent microscopy with DAPI staining was used to examine the cell morphological changes after E11 treatment. DNA fragmentation method was used to detect the inducing effect of emodin derivative E11 on cell apoptosis. Western blot was used to determine the expressions of apoptosis-related proteins including procaspase-9, procaspase-3, PARP and PI3K/AKT, MAPK signalling pathway.

RESULTSEmodin derivative E11 could strongly inhibit the growth of Molt-4 with the IC50 in 48 h at 1.381 ± 0.1552 µmol/L in dose-dependent manner. 0.1 µmol/L of E11 could inhibit cell colony formation. The typrical apopototic morphologic changes of Molt cells treated with E11 could be observed under fluorescence microscope with DAPI staining. DNA apoptotic ladder could be observed by DNA fragmentation.The expressions of procaspase -9, procaspase-3, PARP, p-MAPK, p-AKT, mTOR, p-mTOR, p-P70 and p-4BEP1 were down-regulated, while expressions of MAPK, AKT, 4EBP1 and P70 were not changed remarkably after Molt-4 were treated with E11 for 48 h.

CONCLUSIONE11 can remarkably inhibit the proliferation and induce the apoptosis of Molt-4 cells. The mechanism of apoptosis of Molt-4 cells may be related with the suppression of PI3K/AKT and MAPK signalling pathways.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Down-Regulation ; Emodin ; pharmacology ; Humans ; Leukemia, T-Cell ; pathology ; MAP Kinase Signaling System ; Phosphatidylinositol 3-Kinases ; metabolism ; Poly(ADP-ribose) Polymerases ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; TOR Serine-Threonine Kinases ; metabolism

OBJECTIVETo evaluate the morbidity, risk factors, clinical characterisitics, treatments and prognosis of delayed hepatic veno-occlusive disease(HVOD) after haploidentical hematopoietic stem cell transplantation (hi-HSCT).

METHODSThe clinical data of 208 patients undergoing hi-HSCT were retrospectively analyzed.

RESULTSSix patients were diagnosed with delayed VOD, among them 4 patients were moderate VOD and 2 patients were severe VOD. The incidence of VOD after hi-HSCT was 2.88%, the median onset time was 44.5(30-57) days after transplant, 2 patients died of multiple organ failure (MOF) due to rapid progress of disease. With intravenous administration of defibrotide, 4 patients displayed encouraging response, but 2 patients died of hepatic acute graft-versus-host disease (aGVHD), 1 had bone marrow relapse and the other one was cured.

CONCLUSIONNorethindrone is one of the high risk factors, while sex, age and disease status are irrelevant to the occurrence of VOD. Unfractionated heparin (UH) can effectively decrease the morbidity. Pretransplant hepatic function reserve, high dose preconditioning regimens and pharmacotherapy may result in delayed VOD onset. The delayed VOD has the same clinical features and treatment-response as early VOD, but a poorer prognosis is usually observed. A larger amount of samples (patients) is needed to research the relationship of the delayed VOD with hi-HSCT. Defibrotide can effectively increase the survival rate of VOD patients.

Graft vs Host Disease ; Haploidy ; Hematopoietic Stem Cell Transplantation ; Heparin ; Hepatic Veno-Occlusive Disease ; Humans ; Incidence ; Polydeoxyribonucleotides ; Retrospective Studies ; Risk Factors