Molecular characterization of two leptin genes and their transcriptional changes in response to fasting and refeeding in Northern snakehead (Channa argus).

Leptin has been proved to play critical roles in energy metabolism, body weight regulation, food intake, reproduction and immunity in mammals. However, its roles are still largely unclear in fish. Here, we report two leptin genes (lepA and lepB) from the Northern snakehead (Channa argus) and their transcriptions in response to different feeding status. The snakehead lepA is 781 bp in length and contains a 480 bp open reading frame (ORF) encoding a 159-aa protein, while the snakehead lepB is 553 bp in length and contains a 477 bp ORF encoding a 158-aa protein. Multi-sequences alignment, three-dimensional (3D) model prediction, syntenic and genomic comparison, and phylogenetic analysis confirm two leptin genes are widely existing in teleost. Tissue distribution revealed that the two leptin genes exhibit different patterns. In a post-prandial experiment, the hepatic lepA and brain lepB showed a similar transcription pattern. In a long-term (2-week) fasting and refeeding experiment, the hepatic lepA and brain lepB showed a similar transcription change pattern induced by food deprivation stimulation but differential changes after refeeding. These findings suggest snakehead lepA and lepB are differential both in tissue distribution and molecular functions, and they might play as an important regulator in energy metabolism and food intake in fish, respectively.

Lidocaine promotes fibroblast proliferation after thermal injury via up-regulating the expression of miR-663 and miR-486.

Thermal burn is a complex injury that induces pronounced inflammatory reactions and destruction of the microvasculature. Lidocaine, which is broadly used for pain relief, has been reported have the capacity to modulate wound healing processes. Seventeen burn injury patients with no treatment and nine controls were included in this study. The expression of 15 candidate miRNAs in the dermis samples were detected by qRT-PCR. The target genes were predicted using online bioinformatics tools and confirmed by dual luciferase assay and immunoblotting. The function of miR-486 and miR-663 on skin fibroblasts keratinocytes were determined by MTT assay and flow cytometry. The level of miR-486 and miR-663 was increased after burn injury, meanwhile, the highest level of miR-486 and miR-663 was found after lidocaine treatment. We identified that BCL2L14 was a direct target of both miR-486 and miR-663. Meanwhile, overexpression of miR-486 or miR-663 inhibit apoptosis and promoted proliferation of human skin fibroblasts keratinocytes. These results indicated that lidocaine treatment promoted the skin healing after thermal injury through up-regulating miR-486 and miR-663 expression, and partially explained how lidocaine modulates wound healing processes. This may provide an evidence for the therapeutic effect of lidocaine during skin healing process. However, the underlying mechanisms need to be further unveiled.

Detection of Anaplasma platys in dogs using real-time loop-mediated isothermal amplification.

Anaplasma platys is a parasite of canine platelets that causes infectious cyclic thrombocytopenia. In this study, a novel real-time loop-mediated isothermal amplification (RT-LAMP) method was developed to detect A. platys. RT-LAMP primer sets were designed using a citrate synthase gene sequence and the assay was performed at 63 °C for 30 min. No cross-reactivity was observed with other Anaplasma or Ehrlichia spp. and the method exhibited a similar level of sensitivity in detecting the organism in 58 canine blood samples to that of a nested PCR. This RT-LAMP is a rapid and potentially cost-effective method of diagnosing A. platys infection in dogs.

Quadrilobed superior gluteal artery perforator flap for sacrococcygeal defects.

BACKGROUND: Perforator flaps are used extensively in repairing soft tissue defects. Superior gluteal artery perforator flaps are used for repairing sacral defects, but the tension required for direct closure of the donor area after harvesting of relatively large flaps carries a risk of postoperative dehiscence. This research was to investigate a modified superior gluteal artery perforator flap for repairing sacrococcygeal soft tissue defects. METHODS: From June 2003 to April 2010, we used our newly designed superior gluteal artery perforator flap for repair of sacrococcygeal soft tissue defects in 10 patients (study group). The wound and donor areas were measured, and the flaps were designed accordingly. Wound healing was assessed over a follow-up period of 6 - 38 months. From January 1998 to February 2003, twelve patients with sacrococcygeal pressure sores were treated with traditional methods, VY advancement flaps or oblong flaps, as control group. RESULTS: After debridement, the soft tissue defects ranged from 12 cm × 10 cm to 26 cm × 22 cm (mean 16.3 cm × 13.5 cm). Four patients were treated using right-sided flaps ranging from 15 cm × 11 cm to 25 cm × 20 cm (mean 18.2 cm × 14 cm). Four patients were treated using left-sided flaps, and two were treated using both right- and left-sided flaps. Suction drains were removed on postoperative Days 3 - 21 (mean 5.9) and sutures were removed on postoperative Days 12 - 14. Each flap included 1 - 2 perforators for each of the donor and recipient sites. Donor sites were closed directly. All flaps survived. In eight patients, the wounds healed after single-stage surgery. After further debridement, the wounds of the remaining two patients were considered healed on postoperative Days 26 and 33, respectively. The rate of first intention in the study group (80%, 8/10) significantly increased than that of control group ((25%, 3/12), χ(2) = 4.583, P = 0.032). Follow-up examinations found that the flaps had a soft texture without ulceration. In the two patients without paraplegia, the range of motion of the hip joints was not affected. CONCLUSION: The use of the quadrilobed superior gluteal artery perforator flap can overcome the disadvantages of traditional perforator flaps and represents an improved approach for repairing soft tissue defects in the sacrococcygeal region.

Avian-origin H3N2 canine influenza virus circulating in farmed dogs in Guangdong, China.

Since 2006, more and more cases of the infectious H3N2 canine influenza virus (CIV) in pet dogs have been reported in southern China. However, little is known about the prevalence situation of H3N2 CIV infections in farmed dogs in China. This is the first systematic epidemiological surveillance of CIV in different dog populations in southern China. Two virus strains A/Canine/Guangdong/1/2011(H3N2) and A/canine/Guangdong/5/2011(H3N2) were isolated from canine nasal swabs collected at one dog farm in Guangzhou and the other farm in Shenzhen. Sequence and phylogenetic analysis of eight gene segments of these viruses revealed that they were most similar to the newly isolated canine H3N2 viruses in dogs and cats from Korea and China, which originated from avian strain. This indicates that H3N2 CIV may be a common pathogen for pet and farmed dog populations in southern China at present. Serological surveillance has shown that the infection rate of this avian-origin canine influenza in farmed dogs and in pet dogs were 12.22% and 5.3%, respectively; as determined by the ELISA. The data also suggested that transmission occurred, most probably by close contact, between H3N2 CIV infected dogs in different dog populations in recently years. As H3N2 outbreaks among dogs continue in the Guangdong province (located very close to Hong Kong), the areas where is densely populated and with frequent animal trade, there is a continued risk for pets H3N2 CIV infections and for mutations or genetic reassortment leading to new virus strains with increased transmissibility among dogs. Further in-depth study is required as the H3N2 CIV has been established in different dog populations and posed potential threat to public health.

Oxidative stress parameters and anti-apoptotic response to hydroxyl radicals in fish erythrocytes: protective effects of glutamine, alanine, citrulline and proline.

The present study explored the protective effects of glutamine (Gln), alanine (Ala), citrulline (Cit) and proline (Pro) on hydroxyl radical (·OH)-induced apoptosis in isolated carp erythrocytes. Hydroxyl radicals were generated by ferrous ion (Fe(2+))-mediated decomposition of hydrogen peroxide (H(2)O(2)) (Fenton reaction). In order to select an optimal ·OH concentration to induce apoptosis, cultures were treated with different concentrations of FeSO(4)/H(2)O(2) (0 µM/0 µM-50 µM/25 µM). The results showed that exposure to FeSO(4)/H(2)O(2) (0 µM/0 µM-40 µM/20 µM) increased apoptosis in a dose-dependent manner. Moreover, apoptosis was at its highest level at 40 µM FeSO(4)/20 µM H(2)O(2). We then examined the cytoprotective effects of Gln, Ala, Cit, Pro or the combination of Ala, Cit and Pro under conditions of apoptosis. Carp erythrocytes were treated with the substances listed above in the presence of 40 µM FeSO(4)/20 µM H(2)O(2) for 9 h. The controls were grown in Gln, Ala, Cit, Pro-free culture medium. The results showed that Gln, Ala, Cit, Pro and the combination of Ala, Cit and Pro effectively protected against annexin binding, decrease of forward scatter and DNA fragmentation in carp erythrocytes induced by ·OH. Furthermore, Gln, Ala, Cit, Pro and the combination of Ala, Cit and Pro effectively blocked ·OH-stimulated erythrocyte hemolysis, reduced the increase of superoxide anion and H(2)O(2) concentrations, inhibited the formation of malondialdehyde, protein carbonyls and met-hemoglobin, and prevented the decrease of superoxide dismutase, catalase and glutathione peroxidase activities and glutathione content in carp erythrocytes induced by ·OH. In addition, the results suggest that the combination of Ala, Cit and Pro produces a greater anti-apoptotic and anti-oxidative effect than their individual effects at the same concentrations. Taken together, the results showed that ·OH induces apoptosis and oxidative damage in carp erythrocytes. In addition to inhibiting apoptosis, Gln, Ala, Cit, Pro and the combination of Ala, Cit and Pro protected carp erythrocytes against oxidative damage induced by ·OH, which may be a major factor in the protection of erythrocytes from apoptosis.

[Repair of pressure sores over ischial tuberosity with long head of biceps femoris muscle flap combined with semi-V posterior thigh fasciocutaneous flap].

OBJECTIVE: To explore the clinical effect of transplantation of the long head of biceps femoris muscle flap in combination with semi-V posterior thigh fasciocutaneous flap for repair of pressure sores over ischial tuberosity. METHODS: Eight patients with 10 deep pressure sores over ischial tuberosity were admitted to the First Affiliated Hospital to the PLA General Hospital and the 98th Hospital of PLA from April 2004 to June 2010. The wounds measured from 2 cm × 2 cm to 6 cm × 4 cm were covered with the long head of biceps femoris muscle flap and semi-V posterior thigh fasciocutaneous flap (ranged from 10 cm × 6 cm to 13 cm × 8 cm). The condition of flaps was observed and followed up for a long time. RESULTS: All flaps survived. Nine wounds healed by first intention. Subcutaneous accumulation of fluids occurred in one wound with formation of a sinus at drainage site, and it healed after dressing change for 25 days. Patients were followed up for 7 to 34 months. Sore recurred in one patient 9 months after surgery, and it was successfully repaired with the same flap for the second time. Flaps in the other 7 patients appeared satisfactory with soft texture and without ulceration. CONCLUSIONS: This combined flap is easy in formation and transfer, and it causes little side injury with good resistance against pressure. It is a new method for repair of pressure sore over sacral region.