Biomarkers, Tumor ; metabolism ; Carcinoma, Small Cell ; genetics ; metabolism ; pathology ; Carcinoma, Squamous Cell ; pathology ; Chromosome Aberrations ; Diagnosis, Differential ; Humans ; Keratins ; metabolism ; Lymphoma ; pathology ; Mucin-1 ; metabolism ; Urinary Bladder ; pathology ; Urinary Bladder Neoplasms ; genetics ; metabolism ; pathology

OBJECTIVETo evaluate the effects of Attractin (Atrn) silence on the anti-oxidative and anti-apoptotic abilities of TM4 Sertoli cells and its influence on the expressions of superoxide dismutase (SOD) and caspase6 in the cells.

METHODSWe observed the apoptotic indexes of TM4 Sertoli cells with normal expression (control), partial deletion, and complete deletion of the Atrn gene (psiRNA-TM4, psiAtrn-TM4, and mu-SC). We determined the mRNA and protein expressions of SOD and caspase6 by Q-PCR and Western blot, measured the SOD activity and malondialdehyde (MDA) contentby spectrophotometry, and detected the apoptotic index of the cells by TUNEL.

RESULTSCompared with psiRNA-TM4, after inhibition of the Atrn expression, the Sertoli cells in the psiAtrn-TM4 and mu-SCgroups showed significantly decreased expressions ofSOD mRNA (70.76% and 92.58%) and protein (65.11% and 71.0%) (both P < 0.05). The levels of caspase 6 mRNA and protein were increased 5.28 and 3.40 times in the psiAtrn-TM4 and 2.97 and 2.50 times in the mu-SCgroup as compared with the normal control (both P < 0.05). Atrn deletion markedly increased the apoptotic indexes of the cells in the psiAtrn-TM4 and mu-SC groups by 16.22% and 22.03% (P < 0.05) and reduced the activity of SOD by 23.00% and 39.37% (P < 0.05); it also elevated the level of MDA by 155.22% (P < 0.05).

CONCLUSIONThe Atrn gene exerts influence on the function of Sertoli cells in multiple ways, in which antioxidative stress and apoptosis regulation may play an important role.

Animals ; Apoptosis ; Caspase 6 ; metabolism ; Cells, Cultured ; Gene Deletion ; Male ; Membrane Proteins ; genetics ; metabolism ; Mice ; Oxidative Stress ; Sertoli Cells ; metabolism ; pathology ; Superoxide Dismutase ; metabolism

Carcinoma in Situ ; diagnosis ; pathology ; surgery ; Carcinoma, Transitional Cell ; diagnosis ; pathology ; surgery ; Cystectomy ; Humans ; Neoplasm Invasiveness ; Neoplasm Staging ; methods ; Urinary Bladder ; pathology ; surgery ; Urinary Bladder Neoplasms ; diagnosis ; pathology ; surgery

Aneuploidy ; Gene Rearrangement ; Glutathione S-Transferase pi ; metabolism ; Humans ; Male ; Methylation ; Neoplasm Grading ; Neoplasm Staging ; Neoplasms, Multiple Primary ; genetics ; metabolism ; pathology ; surgery ; Oncogene Proteins, Fusion ; genetics ; Prostate-Specific Antigen ; metabolism ; Prostatic Intraepithelial Neoplasia ; genetics ; pathology ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; surgery

Antineoplastic Agents ; therapeutic use ; Apoptosis ; drug effects ; Caspases ; metabolism ; physiology ; Enzyme Activation ; Fas Ligand Protein ; metabolism ; Humans ; NF-kappa B ; metabolism ; Neoplasms ; metabolism ; therapy ; Neovascularization, Pathologic ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Signal Transduction

The expression of Attractin mRNA and protein in testis and semen of human and male mice was investigated. Human testis and semen samples were all collected from Reproductive Center of Renmin Hospital, Wuhan University in December, 2012. Testis samples were collected from 7 cases of obstructive azoospermias when they were subjected to diagnosed testis biopsy, and 30 normal human semen samples were obtained from those cases of semen analysis. Adult mice testis tissues were obtained from 10 2-month-old male BALB/c mice, and 60 male mice at different ages were classified into 10 groups (day 1, 5, 10, 15, 21, 28, 35, 42, 56, and 120 respectively, n=6 each). The expression of Attractin mRNA and protein in testis was detected by RT-PCR and Western blotting respectively. Human semen samples were centrifuged into sperm plasma (SP) and sperm extract (SE), and mice sperm samples were collected from the epididymis of 10 adult male BALB/c mice. Western blotting was used to determine the Attractin protein expression level. Attractin mRNA and protein were expressed in the testis of both patients with obstructive azoospermias and adult Bcl/B mice. Quantitative RT-PCR revealed that no Attractin mRNA was detectable in day 1 male BALB/c mice group. The Attractin mRNA and protein levels were low on the day 10, and increased with age until day 56. On the day 120, the expression levels of Attractin were decreased. As for human semen samples, Attractin protein was expressed in both SP and SE, but didn't exist in samples from the epididymis of male BALB/c mice. It was suggested that Attractin acted as a novel active substance and was involved in male reproduction in both human and BALB/c mice, but it exerted a different expression profile in different mammal species.

This paper is aim to investigate the pharmacokinetics and absolute bioavailability of neoline in Beagle dogs, and provide a theoretical basis for further study. Ethyl acetate was used for liquid-liquid extracting after 10% ammonia alkalizing. The method of UPLC-Q-TOF-MS was established for the determination of neoline plasma concentrations. Beagle dogs were orally or intravenously administered with neoline for pharmacokinetic and absolute bioavailability study. Good linear relationship of neoline was found over the range of 0.1-4 mg x L(-1) (R2 = 0.9982) and 2-100 microg x L(-1) (R2 = 0.9945). Intra-and inter-day precision, expressed as the relativestandard (RSD) were less than 5.0%. Accuracy, expressed as the relative error (RE) was within 90.0%-115%. The recovery of neoline in dog plasma was more than 80%. After 6 mg x kg(-1) for ig and 1 mg x kg(-1) for iv administration of neoline, the main pharmacokinetic parameters were analyzed with Winnonlin software. t(1/2) were (313.88 +/- 63.18), (236.33 +/- 229.84) min, and AUC(0-infinity) were (58,027.40 +/- 14,132.69), (473,578.02 +/- 82,333.08) min x microg x L(-1) for ig and iv administration respectively. The absolute bioavail ability was (73.15 +/- 10.29) %. The method of UPLC-Q-TOF-MS described in the report was sensitive, reliable and specific, and suitable for pharmacokinetic study of neoline in Beagle dog. The high absolute bioavailability of neoline in dog suggested good absorption of neline which was worth of further investigation.
Aconitine ; analogs & derivatives ; chemistry ; pharmacokinetics ; Animals ; Biological Availability ; Dogs ; Drug Stability ; Female ; Male

OBJECTIVETo evaluate the functional outcomes of modified limited "L" incision beside the Achilles tendon with distraction bone block arthrodesis in treatment of subtalar osteoarthritis.

METHODSFrom March 2009 to September 2012, a total of 22 cases of old calcaneus fractures with subtalar osteoarthritis were treated with modified limited "L" incision and distraction bone block arthrodesis including 13 males and 9 females with a mean age of 35.3 years old (ranged 22 to 49). The mean time from calcaneal fracture was 21 months (ranged 11 to 32). According to the Stephens-Sanders classification, 16 cases were type II and 6 were type III. The modified-AOFAS ankle-hindfoot score was used for functional outcomes evaluation.

RESULTSThere was one incision necrosis and no infection, implant failure, bone-graft absorbed or talus necrosis was note at the follow-up time. A total of 21 cases were followed up for a mean time of 29 months (ranged from 18 to 46 months). All of the cases reached a bony union within 4 months postoperation. The mean modified-AOFAS ankle-hindfoot score was 82.6 points (ranged from 66 to 92 points),reached a significantly improvement in comparing with the mean preoperative score (50.8 points,ranged from 32 to 65 points, P < 0.01).

CONCLUSIONThe modified limited"L" incision beside the Achilles tendon with distraction bone block arthrodesis is an acceptable and alternative treatment method for subtalar osteoarthritis. This method is easy to use and with less complication. It can correct the main pathological changes and reach good functional outcomes.

Adult ; Arthrodesis ; methods ; Female ; Humans ; Male ; Middle Aged ; Osteoarthritis ; surgery ; Subtalar Joint ; surgery

This study aimed to examine the diagnosis performance of 99mTc-methoxyisobutylisonitrisonitrile (99mTc-MIBI) and multimodality imaging [ultrasound,single-photon emission computed tomography/computed tomography (SPECT/CT)] for hyperparathyroidism (HPT).From Nov.2009 to Dec.2015,clinical data of a total of 43 HPT patients (16 males and 27 females;26-70 years old,average age:51.60±10.66 years old) were retrospectively analyzed.Among them,19 patients with primary hyperparathyroidism (PHPT) underwent 99mTc-MIBI planar imaging,24 [15 with PHPT and 9 with secondary hyperparathyroidism (SHPT)] underwent SPECT/CT hybrid imaging,and 41 (33 with PHPT and 8 with SHPT) had neck ultrasound imaging.Final diagnosis was determined by pathological examination after surgery.The positive rate was compared between different imaging modalities,and the correlation analysis was conducted between imaging results and lesion size or serum parathyroid hormone (PTH) level.The results showed that the total positive rates of 99mTc-MIBI imaging,ultrasound,and the two combined imaging in the 43 HPT cases were 90.70％ (39/43),58.54％ (24/41),and 100％ (41/41),respectively.According to lesion numbers,the positive rates were 79.10％ (53/67),53.23％ (33/62),and 88.71％ (55/62),respectively.SPECT/CT hybrid images were positive in all the 24 patients who underwent this examination.The mean maximum diameters of the lesions in 99mTc-MIBI positive and negative patients were 1.96±0.95 cm and 1.36±0.67 cm respectively,with statistically significant difference noted (P=0.03).The T/NT of 99mTc-MIBI imaging at the early phase was correlated positively with serum PTH level (r=0.40,P=0.01).The T/NT of 99mTc-MIBI imaging at both the early phase and the delay phase was correlated positively with lesion size (r=0.51,and r=0.45,respectively;P＜0.01 for both).It was concluded that 99mTc-MIBI imaging presents significant value for location diagnosis of HPT,especially when combined with SPECT/CT hybrid imaging or ultrasound.The 99mTc-MIBI uptake correlates positively with serum PTH level and lesion size.

We analyzed genetic evolution characteristics of avian influenza A (H7N9) virus isolated in Zhaoqing,China,2014-2016.Nucleic acid were extracted and sequenced from 17 samples of H7N9 positive cases in Zhaoqing.Genetic characteristics of homology and important amino acid sites were analyzed by using BioEdit5.0 and MEGA6.0.The evolutionary trees were constructed by Neighbor-Joining and the referenced sequences were downloaded from GenBank,Eight nucleic acid fragments from 7 strains of H7N9 viruses were successfully generated.The highest homology was found in HA gene with A/chicken/Dongguan/695/2014(H7N9),and NA gene with A/chicken/Dongguan/1075/2014(H7N9).The internal genes were high homology with avian H7N9 and H9N2 virus from Dongguan and Shenzhen in Guangdong,China.The HA and NA genes were directly evolved in the Pearl River Delta evolution branch with the H7N9 sequences from the cities of Dongguan,Guangzhou and Shenzhen,while the sequences from the provinces of Anhui,Zhejiang,and Jiangsu were in the Yangtze River Delta evolution branch.There were 2 alkaline amino acids in cleavage site of HA,2 mutations (G186V and Q226L) in the crucial sites related with the receptor of HA protein,1 mutation (E627K) in PB2 protein,and 1 drug resistance mutation (S31N) in M2 protein.And no evidence of neuraminidase resistance in NA protein was found.In conclusion,the H7N9 virus for human infection in Zhaoqing may originate from avian H7N9 and H9N2 viruses,which circulated in the Pearl River Delta region of Guangdong from 2013 to 2014.The mutations of G186V,Q226L and E627 K might be related with high susceptibility to human beings.