A new odorous frog species of Odorrana (Amphibia, Anura, Ranidae) from Guizhou Province, China.

The frog genus Odorrana is distributed across east and southeastern Asia. Based on morphological differences and molecular phylogenetics, a new species of the genus occurring from Leigong Mountain in Guizhou Province, China is described. Phylogenetic analyses based on DNA sequences of the mitochondrial 12S rRNA, 16S rRNA, and ND2 genes supported the new species as an independent lineage. The uncorrected genetic distances between the 12S rRNA, 16S rRNA, and ND2 genes between the new species and its closest congener were 5.0%, 4.9%, and 16.3%, respectively. The new species is distinguished from its congeners by a combination of the following characters: body size moderate (SVL 39.1-49.4 mm in males, 49.7 mm in female); head width larger than head length; tympanum distinctly visible; small rounded granules scattered all over dorsal body and limbs; dorsolateral folds absent; heels overlapping when thighs are positioned at right angles to the body; tibiotarsal articulation reaching the level between eye to nostril when leg stretched forward; vocal sacs absent in male and nuptial pads present on the base of finger I.

Protective effects of fowl-origin cadmium-tolerant lactobacillus against sub-chronic cadmium-induced toxicity in chickens.

Cadmium (Cd) directly endangers poultry health and indirectly causes harm to human health by food chain. Numerous studies have focused on removing Cd using lactic acid bacteria (LAB). However, there is still a lack of in vivo studies to validate whether Cd can be absorbed successfully by LAB to alleviate Cd toxicity. Here, we aimed to isolated and screened poultry-derived Cd-tolerant LAB with the strongest adsorption capacity in vitro and investigate the protective effect of which on sub-chronic Cd toxicity in chickens. First, nine Cd-tolerant LAB strains were selected preliminarily by isolating, screening, and identifying from poultry farms. Next, four strains with the strongest adsorption capacity were used to explore the influence of different physical and chemical factors on the ability of LAB to adsorb Cd as well as its probiotic properties in terms of acid tolerance, bile salt tolerance, drug resistance, and antibacterial effects. Resultantly, the CLF9-1 strain with the best comprehensive ability was selected for further animal protection test. The Cd-tolerant LAB treatment promoted the growth performance of chickens and reduced the Cd-elevated liver and kidney coefficients. Moreover, Cd-induced liver, kidney, and duodenum injuries were alleviated significantly by high-dose LAB treatment. Furthermore, LAB treatment also increased the elimination of Cd in feces and markedly reduced the Cd buildup in the liver and kidney. In summary, these findings determine that screened Cd-tolerant LAB strain exerts a protective effect on chickens against sub-chronic cadmium poisoning, thus providing an essential guideline for the public health and safety of livestock and poultry.

Epidemiological features of lung giant cell carcinoma and therapy for patients with EGFR mutations based on case reports and the surveillance, epidemiology, and end results (SEER) database.

Epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) are the standard first line treatment for advanced non-small cell lung cancer (NSCLC) with sensitive EGFR mutations. Among NSCLC, giant cell carcinoma of the lung (GCCL) is a rare pathological subtype with poor prognosis, with no confirmed evidence about its epidemiological features or therapeutic efficiency of EGFR-TKIs. We present two advanced GCCLs with sensitive EGFR mutations, also collected the cases of GCCL from our hospital and the Surveillance, Epidemiology, and End Results (SEER) program. Kaplan-Meier methods and Cox proportional hazards modeling were used to perform the survival analyses. Both two cases of advanced GCCL with sensitive EGFR mutations benefited from EGFR-TKIs. Twelve GCCLs were recorded in our hospital from May 2006 to July 2015. GCCL is associated with males (83.3%) and smoking status (63.6%). The EGFR mutation rate was 40.0%. In SEER database, the total number of GCCLs was 184, 0.11% for all NSCLCs. In Kaplan-Meier analysis, the 5-year overall survival of GCCL patients was significantly lower than that of non-GCC NSCLC (16% and 19%; P<0.001), and it was confirmed in multivariate analysis. Further survival analyses indicated that male were more susceptible to GCCL and GCCL was prone to metastasize. Only age and M stage were independent prognostic factors for GCCL in the multivariate analysis. In conclusion, GCCL was an unfavorable prognostic factor and associated with males and metastasis. GCCL patients with sensitive EGFR mutations may also benefit from EGFR-TKI, we therefore recommend the evaluation of EGFR in the treatment of advanced GCCL.

The genetic diversity of chicken breeds from Jiangxi, assessed with BCDO2 and the complete mitochondrial DNA D-loop region.

The Jiangxi Province of China has numerous native domestic chicken breeds, including some black skin breeds. The genetic diversity of Jiangxi native chickens is largely unknown, and specifically, the genetic contribution of the grey junglefowl to black skin chickens is not well understood. To address these questions, the complete D-loop region of the mitochondrial DNA (mtDNA) and beta-carotene dioxygenase 2(BCDO2)gene was sequenced in a total of 209 chickens representing seven Jiangxi native breeds. Thirty-one polymorphic sites were identified across the complete mtDNA D-loop region sequence. Twenty-three haplotypes were observed in the seven breeds, which belonged to four distinct mitochondrial clades (A, B, C and E). Clade A and B were dominant in the chickens with a frequency of approximately 67.9%. There were five SNPs that defined two haplotypes, W and Y in BCDO2. Four breeds had one haplotype and three breeds had two. We conclude that Jiangxi native chicken breeds have relatively low genetic diversity and likely share four common maternal lineages from two different maternal ancestors of junglefowl. Furthermore, some Jiangxi chicken populations may have been mixed with chickens with exotic lineage. Further research should be established to protect these domestic chicken resources.

The investigation of genetic diversity and evolution of Daweishan Mini chicken based on the complete mitochondrial (mt)DNA D-loop region sequence.

This study evaluated the genetic diversity and origin of Daweishan Mini chickens using mtDNA sequence polymorphism. Blood samples from 30 Daweishan Mini chickens were collected. The complete D-loop was PCR amplified, sequenced and compared with the DNA data of five Red Junglefowl (Gallus gallus) subspecies. Eighteen variable sites that defined six haplotypes were observed. The six haplotypes were clustered into four clades (A, B, D and E), of which clade A and B were dominant. Clades Aand B were clustered with G.g. spadiceus, indicating these two clades may have originated from this subspecies. These results show there is diversity in the middle of the mtDNA D-loop, and indicate there are multiple maternal origins for Daweishan Mini chickens. It appears that G.g. spadiceus contributed more to the evolution of the Daweishan Mini chickens breed than the other four subspecies tested here.

Reconstruction of the Achilles tendon and overlying skin defect: 3 case reports.

Although various reconstruction surgery techniques are available to repair posterior heel defects, the compound defects reconstruction is an ongoing surgical challenge. Complex, free tissue flaps are often clinically used in this repair operation but the techniques have some disadvantages, including intraoperative tedious dissections, vascular anastomosis, and postoperative thrombogenesis. Here, we present a single-stage procedure for Achilles tendon and its overlying skin defects repair with a complex posterior tibial artery perforator-based tissue flap on 3 patients. This method can repair the Achilles tendon and the soft tissue defects simultaneously in a relatively short operative time. The prognosis of the 3 operative patients described here was great for participating in exercise and daily work unassisted 18 to 26 months after operation. Clinical results indicate that our operative method can be effective in repair of Achilles tendon and its overlying skin defects without major complications.

[Effects of different ways in repairing scrotum of pigs with full-thickness burn on apoptosis and bcl-2 protein expression of spermatogenic cells].

OBJECTIVE: To observe the effects of different ways in repairing scrotum of pigs with full-thickness burn on spermatogenesis of testis. METHODS: Twenty male Guizhou miniature pigs were divided into normal control (NC), natural-healing (NH), flap-repairing (FR), and skin-grafting (SG) groups according to the random number table, with 5 pigs in each group. Pigs in NC group were not subjected to any injury. Scrotum of pigs in the latter three groups were inflicted with full-thickness burn. Wounds in NH group healed naturally. Wounds in FR group were repaired with inguinal region flap, and those in SG group with full-thickness skin from lower abdomen. Appearance of scrotum in the latter three groups was observed right after injury, and three months post injury or surgery (PIM or PSM). Specimens of testes of pigs in the latter three groups were obtained in PIM or PSM 3 to detect apoptosis of spermatogenic cells with TUNEL, and bcl-2 protein expression with immunohistochemistry. The same indexes were observed and determined in pigs of NC group. Data were processed with one-way analysis of variance and LSD test. RESULTS: (1) Scrotum of pigs in NC group had skin folds with contraction function. Scrotum of pigs became hard with a leathery appearance right after burn in the other three groups. In PIM or PSM 3, wounds of pigs in NH group healed with scar, and the testes were squeezed into inguinal region. Scrotal skin of pigs in FR group was thick with testes in the scrotum, and that of pigs in SG group was thin with testes in the scrotum. (2) Spermatogenic cells in each level in NC group were arranged regularly, with few apoptotic spermatocytes and spermatoblasts. In NH, FR, and SG groups, seminiferous epithelium was thinner with most of the spermatogenic cells showing apoptosis, and they were mainly spermatogonia and spermatocytes. Apoptotic index of spermatogenic cells in NH, FR, SG, and NC groups was respectively (46.3 ± 3.3)%, (40.9 ± 3.5)%, (20.6 ± 2.3)%, (7.5 ± 1.9)%, and the difference among them was statistically significant (F = 405.65, P < 0.01). There were significant statistical differences among the former three groups (with P values below 0.01). (3) bcl-2 protein expression in NH, FR, SG, and NC groups was respectively (52 ± 5)%, (53 ± 4)%, (64 ± 5)%, (75 ± 5)%, and the difference among them was statistically significant (F = 56.63, P < 0.01). There was no significant statistical difference in bcl-2 expression between NH group and FR group (P = 0.66), and it was lower in both groups as compared with SG group (with P values below 0.01). CONCLUSIONS: Either scar healing, flap transplantation, or SG in repairing scrotum with full-thickness burn in pigs inhibits spermatogenesis, but repair with SG produces less deleterious effect on the testis.

[Impact of the reconstruction of the scrotum with third-degree burns on spermatogenic function: experimental study of three different methods].

OBJECTIVE: To explore the effects of different methods of scrotal reconstruction on the apoptosis of spermatogenic cells and expression of the bcl-2 protein in patients with third-degree scrotal burns. METHODS: Forty male and 24 female 2-month-old Guizhou mini-pigs were used in this study, the former equally randomized to groups I (normal control), II (natural healing), III (skin grafting) and IV (skin flap grafting). Ten months after the establishment of the model of third-degree burns, 6 male pigs from each group were paired with the female pigs and fed for 3 weeks. Then the female pigs were fed for another 4 months, followed by observation of their reproductivity. At 12 months, the bilateral testes were taken from the male pigs for detection of the apoptosis index of spermatogenic cells by TUNEL and determination of the expression of the bcl-2 protein by immunohistochemistry. The data obtained were subjected to single factor analysis of variance. RESULTS: The apoptosis indexes of spermatogenic cells were (7.07 +/- 3.5), (40.34 +/- 4.85), (15.14 +/- 1.36) and (39.29 +/- 5.73)% in groups I , II, III and IV, respectively, significantly higher in groups II , III and IV than in I (P<0.05), with statistically significant differences between group III and groups II and IV (P<0.05) but not between II and IV (P>0.05). The expression rates of the bcl-2 protein were (75.07 +/- 3.74), (54.93 +/- 4.03), (66.85 +/- 3.06) and (53.33 +/- 5.22)% in groups I, II, III, and IV, respectively, remarkably higher in I than in the other three (P<0.05), with significant differences between group III and groups II and IV (P<0.05) but not between II and IV (P>0.05). Pregnancies were found in all the female pigs of group I with 10.0 +/- 1.18 newborns and in 4 of group III with 9.92 +/- 1.31 newborns, but in none of groups II and IV, with significant differences between group I and the other three (P<0.05) as well as between group III and groups II and IV (P<0.05), but not between II and IV (P>0.05). CONCLUSION: All the three methods of reconstruction for the scrotum with third-degree burns can suppress spermatogenic function, increase the apoptosis of spermatogenic cells and decrease the expression of the bcl-2 protein, among which, skin grafting least affects spermatogenic function.

Rapid and sensitive detection of Listeria monocytogenes by loop-mediated isothermal amplification.

Loop-mediated isothermal amplification (LAMP) was designed for detection of Listeria monocytogenes, which is an important food-borne kind of pathogenic bacteria causing human and animal disease. The primers set for the hlyA gene consist of six primers targeting eight regions on specific gene. The LAMP assay could be performed within 40 min at 65°C in a water bath. Amplification products were visualized by calcein and manganous ion and agarose gel electrophoresis. Sensitivity of the LAMP assay for detection of L. monocytogenes in pure cultures was 2.0 CFU per reaction. The LAMP assay was 100-fold higher sensitive than that of the conventional PCR assay. Taking this way, 60 chicken samples were investigated for L. monocytogenes. The accuracy of LAMP was shown to be 100% when compared to the "gold standard" culture-biotechnical, while the PCR assay failed to detect L. monocytogenes in two of the positive samples. It is shown that LAMP assay can be used as a sensitive, rapid, and simple detection tool for the detection of L. monocytogenes and will facilitate the surveillance for contamination of L. monocytogenes in food.

[Repair of wounds in popliteal fossa scar with perforator flaps of gastrocnemius medial head].

OBJECTIVE: To study the effect of perforator flaps of medial head of gastrocnemius muscle on repairing popliteal fossa scar wounds. METHODS: Ten patients with scar in popliteal fossa hospitalized from January 2005 to January 2010 were repaired with perforator flaps of medial head of gastrocnemius muscle. The scar was resulted from burn in 8 patients, and from operation in 2. The duration of the scar was 3 months to 11 years, and area of the scar ranging from 6 cm × 3 cm to 10 cm × 6 cm. Ultrasonic Doppler was used to detect the musculocutaneous perforator vessel of the medial sural artery at the position 10 to 17 cm from the fold of the popliteal fossa and 2 to 5 cm from the posterior midline before surgery. Then flap transplantation surgery was performed. Donor site wounds with width less than 5 cm were sutured directly, and those wider than 5 cm were repaired with skin transplantation. RESULTS: All the flaps survived. Flap size ranged from 7 cm × 5 cm to 12 cm × 7 cm. All patients were followed up for 3 to 30 months, and the flaps were found to have a good appearance. Patients could walk with heavy load without lameness. The function of knee joint of the affected limb was the same as that of the opposite limb. No obvious depression was observed in donor sites. CONCLUSIONS: The perforator flaps of medial head of gastrocnemius muscle can be used to repair the popliteal fossa scar wound with satisfactory result.

[Dorsal metatarsal artery distal perforator pedicle link-pattern flaps for wound repairing in the distal feet].

OBJECTIVE: To investigate the feasibility of repairing the wounds in the distal feet with dorsal metatarsal artery distal perforator pedicle link-pattern flaps. METHODS: Since January of 2004 to April of 2009, 30 patients with distal wounds in the feet underwent surgical wound repair using by dorsal metatarsal arteries distal perforator pedicle link-pattern flaps. Seventeen patients used the 1, 2 dorsal metatarsal artery distal perforator flaps, and 13 had the 3, 4 dorsal metatarsal artery distal perforator flaps, with the flaps measuring 4 cmx4 cm to 8 cmx7cm and the flap pedicle ranged 3-6 cm in length. The donor sites were repaired with skin grafting. RESULTS: Twenty-nine flaps survived completely and 1 flap showed partial necrosis in the distant part (2.0 cmx1.0 cm). Twenty-one patients were followed up for an average of 11 months, during which the color, texture, and contour of the flaps remained normal without ulcers in the donor sites or the flaps. CONCLUSION: The wounds in the distal feet can be repaired by dorsal metatarsal artery distal perforator pedicle link-pattern flaps, which is a simple, applicable and safe procedure.

[Applied anatomy of medial plantar artery combined flaps and repairing heel and adjacent vast soft tissue defects].

OBJECTIVE: To explore an anatomical basis and incising scope for the medial plantar artery combined flaps and its feasibility of repairing heel and adjacent vast soft tissue defects. METHODS: The origin, branches, course and distribution of the medial plantar artery were studied in 20 legs of adult cadavers. The largest area of healthy adult's medial plantar and medialis pedis scope that could be incised for the combined flap was measured. Ten cases of heel and adjacent vast soft tissue defects were repaired by the combined medial plantar and medialis pedis flaps. Age range 28-60 years old. The areas of flaps were 9 cm x 7 cm - 10 cm x 8 cm. Defects of tendo calcaneus were repaired by the tendon of abductor muscle and plantar fascia in 3 cases with skin grafts on the donor sites. RESULTS: The medial plantar artery gave off deep branch (diameter: 1.5 mm +/- 0.3 mm) and superficial branch (diameter: 1.0 mm +/- 0.2 mm). The length of media plantar artery deep branch was (8.9 +/- 0.2) cm and gave off 3-5 cutaneous branches. The largest area of combined flap was about 10.4 cm x 9.2 cm - 9.1 cm x 8.2 cm. Ten flaps survived completely. Eight flaps were followed up for 1 month to 1.5 years. The color, texture and appearance of flaps were excellent. The flaps of two-point discrimination were not different with opposite medial region. The affected limb exercised freely and had no ulceration on the flap. CONCLUSIONS: The combined flap has a reliable blood supply and it can recover good sensation after operation. It is one of ideal flaps to repair heel and adjacent vast soft tissue defects. Its disadvantage is a complicated operation.

[Repair of skin defects at forepart of feet with reverse saphenous nerve neurocutaneous flaps].

OBJECTIVE: To investigate the effect of reverse saphenous nerve neurocutaneous flaps for skin defects at forepart of feet. METHODS: From January 2004 to October 2008,15 cases of skin defects at forepart of feet were repaired with reverse saphenous nerve neurocutaneous flaps. The flap size ranged from 3.5 cm x 3.0 cm to 8 cm x 5 cm. The wounds at donor site were closed with skin graft. RESULTS: All the flaps survived completely with no ulcer at the donor site. 10 patients were followed up for 1 to approximately 9 months. The skin color and texture were satisfactory. The patients could walk very well. CONCLUSIONS: It is reliable to repair the skin defects at forepart of feet with reverse saphenous nerve neurocutaneous flaps. It is easily performed with less morbidity.

[Fibular artery perforator link-pattern flaps at lateral and posterior part of leg].

OBJECTIVE: To investigate the feasibility of fibular artery perforator link-pattern flaps at lateral and posterior part of legs. METHODS: From January 2007 to March 2009, 12 cases of with feet and ankle wounds were treat with fibular artery perforator link-pattern flaps at lateral and posterior part of legs. The flap size ranged from 10 cm x 10 cm to 25 cm x 13 cm. The wounds at the donor sites were closed with skin grafts. RESULTS: All the flaps survived completely. The patients were followed up for 1-12 months (median, 6 months). The color, texture and appearance of the flaps were good. CONCLUSION: The link-pattern flap has reliable blood supply and a large area for repairing defects. The sural nerve is reserved, resulting minimal morbidity to donor site.