Objective To assess the inlfuence of depth on liver stiffness measurement with real-time shear wave elastography (SWE) and determine the optimal depth for SWE in liver. Methods SWE of liver was performed on 89 healthy volunteers between May 2012 and November 2012. The depths of each liver were varied from 0 cm to 7 cm (from the liver capsule) in 1 cm increment and there were 8 depth groups in total. Then the elastic modulus of liver in each depth group were measured three times by SWE. The body mass index (BMI) and the distance from body surface to liver capsule were documented. The success rates and the mean elastic modulus of each group were calculated. Results The success rates of 0-7 cm were 0, 98.9%(88/89), 98.9%(88/89), 98.9%(88/89), 71.9%(64/89), 24.7%(22/89), 3.4%(3/89) and 0, respectively. The success rates were highest in 1 cm, 2 cm and 3 cm groups but signiifcant decreased with the increasement of depths in 4 cm, 5 cm and 6 cm groups ( 3 cm vs 4 cm, χ2=25.94, P<0.001; 4 cm vs 5 cm, χ2=39.68, P<0.001;5 cm vs 6 cm,χ2=16.79, P<0.001). The mean elastic modulus of 1 cm, 2 cm, 3 cm, 4 cm and 5 cm groups were (4.77±0.99), (4.68±0.99), (4.76±0.95), (5.19±1.10) and (5.41±0.95) kPa, respectively. The mean elastic modulus of 4 cm and 5 cm groups were signiifcant higher than those of 1 cm, 2 cm, 3 cm groups (4 cm vs 1 cm, t=-2.85, P=0.005;4 cm vs 2 cm, t=-3.49, P=0.001;4 cm vs 3 cm, t=-2.76, P=0.006;5 cm vs 1 cm, t=-3.13, P=0.002;5 cm vs 2 cm, t=-3.66, P=0.000;5 cm vs 3 cm, t=-3.05, P=0.003). In the group of 4 cm, the BMI and the distance from body surface to liver capsule of the volunteers performed successfully and unsuccessfully were (20.70±2.87), (22.07±2.42) kg/m2 and (1.45±0.25 ), (1.60±0.29) cm, respectively. In the group of 5 cm, the BMI and the distance from body surface to liver capsule of the volunteers performed successfully and unsuccessfully were (19.82±2.76), (21.49±2.72) kg/m2 and (1.35±0.21), (1.54±0.26) cm respectively. The BMI had no signiifcant difference between the successful and unsuccessful groups (t=-2.83, P=0.108 for 4 cm;t=0.77, P=0.709 for 5 cm), but the distance from body surface to liver capsule was signiifcantly different (t=26.51, P=0.012 for 4 cm;t=79.57, P=0.004 for 5 cm). Conclusions The success rates and elastic modulus were different at different depths. SWE should be performed at the depths of 1-3 cm from the liver capsule.

Action Potentials ; Animals ; Atrial Fibrillation ; physiopathology ; Atrial Function, Left ; physiology ; Cardiac Pacing, Artificial ; Pulmonary Veins ; physiology ; Rabbits ; Streptomycin ; pharmacology ; Verapamil ; pharmacology

Adenosine ; pharmacology ; Animals ; Cyclic Nucleotide-Gated Cation Channels ; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels ; Ion Channels ; physiology ; Isoproterenol ; pharmacology ; Membrane Potentials ; drug effects ; Muscle, Smooth, Vascular ; physiology ; Myocytes, Smooth Muscle ; physiology ; Potassium Channels ; Pulmonary Veins ; physiology ; Rabbits

OBJECTIVETo investigate the RHD zygosity distribution and the genetic characteristics of RHD gene in the Dong ethnic population in southeast area of Guizhou province.

METHODSBased on the characteristics of Chinese RHD and the RHD specific deletion, two pairs of primers specific for hybrid Rhesus box and exon 1 of RHD respectively, were designed, combined with a pair of internal control primers. Polymerase chain reaction was performed to genotype the samples.

RESULTSIn the 292 RhD positive samples, 58 (19.86%) were RHD+/RHD- heterozygotes, and the others (80.14%) were RHD+/RHD+ homozygotes. In the 9 D negative samples, 5 were RHD+/RHD- heterozygotes (2 weak D, 3 Del), 3 were RHD+/RHD+ homozygotes (1weak D, 2 Del), and 1 was RHD-/RHD- homozygote.

CONCLUSIONRHD+/RHD- heterozygosity is higher (19.86%) in D positive individuals of Dong ethnic group in Guizhou province than that in other areas; RHD gene heterozygosity is also high in the D negative individuals in this ethnic group.

China ; Ethnic Groups ; genetics ; Exons ; Gene Deletion ; Genotype ; Heterozygote ; Homozygote ; Humans ; Polymerase Chain Reaction ; methods ; Rh-Hr Blood-Group System ; genetics

In vitro method as indispensable biological research and testing tools, has been widely used in cosmetics compliance test, biological mechanism research and functional materials screening. These methods include alternative methods which have been accepted by reguations and standardized test guide, non-testing method used for risk assessment and prediction, and a lot more diversification and individualization of in vitro methods. With the consensus formation of the whole industry and increase of application experience, innovation and optimization in vitro method will be constantly emerge. These technologies will be beneficial to improve competitiveness and to develop products to meet the needs of consumers.

Objective:To identify a two-component signaling system (TCSS) composed of β-lactam antibiotic resistance-associated intracellular CiaR and transmembrane serine/threonine kinase StkP of Streptococcus pneumoniae ( S. pneumoniae). Methods:The intracellular segment of stkP gene (IC- stkP) was amplified by PCR and the PCR product was sequenced after T-A cloning. A prokaryotic expression system for IC- stkP segment was established. SDS-PAGE was used to detect the expression of the target recombinant proteins (rIC-StkP and rCiaR) by the established prokaryotic expression system and a previously established prokaryotic expression system for ciaR gene. Ni-NTA affinity chromatography was used to purify the recombinant proteins. The rIC-StkP-captured target proteins of S. pneumoniae were identified by LC-MS/MS after Co-IP. The ability of rCiaR to bind to rIC-StkP was detected by surface plasmon resonance (SPR) and isothermal titration calorimetry (ITC). Results:The established prokaryotic expression system for IC- stkP segment could effectively express rIC-StkP. Both rIC-StkP and rCiaR after purification showed a single protein band on SDS-PAGE. CiaR could be specifically co-precipitated with rIC-StkP. Three extracted cleaved peptides were found in CiaR molecule with exactly matched sequences. SPR and ITC showed that rCiaR could strongly bind to rIC-StkP with high affinity and the KD values were 1.526×10 -9 mol/L and 1.980×10 -6 mol/L, respectively. Conclusions:S. pneumoniae CiaR could act as the downstream response regulatory protein of StkP kinase to compose StkP/CiaR TCSS.

Background: Laparoscopic hepatectomy is increasingly being used to treat hepatocellular carcinoma (HCC). How?ever, few studies have examined the treatment of recurrent HCC in patients who received a prior hepatectomy. The present prospective study compared the clinical efcacy of laparoscopic surgery with conventional open surgery in HCC patients with postoperative tumor recurrence. Methods: We conducted a prospective study of 64 patients, all of whom had undergone open surgery once before, who were diagnosed with recurrent HCC between June 2014 and November 2014. The laparoscopic group (n = 31)underwent laparoscopic hepatectomy, and the control group (n tion time, intraoperative blood loss, surgical margins, postoperative pain scores, postoperative time until the patient= 33) underwent conventional open surgery. Opera?could walk, anal exsufation time, length of hospital stay, and inpatient costs were compared between the two groups. The patients were followed up for 1 year after surgery, and relapse?free survival was compared between the two groups. Results: All surgeries were successfully completed. No conversion to open surgery occurred in the laparoscopic group, and no serious postoperative complications occurred in either group. No significant difference in inpatient costs was found between the laparoscopic group and the control group (P = 0.079), but significant differencesbetween the two groups were observed for operation time (116.7 ± 37.5 vs. 148.2 ± 46.7 min, P = 0.031), intraopera?tive blood loss (117.5 ± 35.5 vs. 265.9 ± 70.3 mL, P = 0.012), postoperative time until the patient could walk (1.6 ± 0.6vs. 2.2 ± 0.8 days, P < 0.05), anal exsufation time (2.1 ± 0.3 vs. 2.8 ± 0.7 days, P = 0.041), visual analogue scale pain score (P < 0.05), postoperative hepatic function (P < 0.05), and length of hospital stay (4.5 ± 1.3 vs. 6.0 ± 1.2 days,P= 0.014). During the 1?year postoperative follow?up period, 6 patients in each group had recurrent HCC on the side of the initial operation, but no significant difference between groups was observed in the recurrence rate or relapse?free survival. In the laparoscopic group, operation time, postoperative time until the patient could walk, anal exsufation time, and inpatient costs were not different (P > 0.05) between the patients with contralateral HCC recur?rence (n = 18) and those with ipsilateral HCC recurrence (n = 13). However, intraoperative blood loss was signifi?cantly less (97.7 ± 14.0 vs. 186.3 ± 125.6 mL, P = 0.012) and the hospital stay was significantly shorter (4.2 ± 0.7 vs. 6.1 ± 1.7 days, P = 0.021) for the patients with contralateral recurrence than for those with ipsilateral recurrence. Conclusions: For the patients who previously underwent conventional open surgical resection of HCC, complete laparoscopic resection was safe and effective for recurrent HCC and resulted in a shorter operation time, less intraop?erative blood loss, and a faster postoperative recovery than conventional open surgery. Laparoscopic resection was especially advantageous for the patients with contralateral HCC recurrence.

Objective To analyze the magnetic resonance imaging (MRI) of the spinal cord and clinical characteristics in patients with autoimmune glial fibrillary acidic protein (GFAP) astrocytopathy.Methods A total of 1 040 samples of cerebrospinal fluid (CSF) and sera collected in the Second Affiliated Hospital of Guangzhou Medical University from March 2013 to June 2018 were tested with tissue-and cell-based assays,and 42 patients were found positive for GFAP-IgG.The clinical data and MRI characteristics of the spinal cord of 19 patients who were positive for GFAP-IgG in CSF with autoimmune GFAP astrocytopathy and lesions in the spinal cord were retrospectively reviewed.Results There were 12 females and seven males among the 19 patients,with onset age of (44±17) years.The main manifestations of these patients included limb weakness (14/19),abnormal vision (5/19),headache (4/19),seizure (4/19),dementia (3/19),etc.On MRI of the spinal cord,five patients showed involvement in the cervical cord alone,eight showed involvement in the thoracic cord alone and six had both cervical and thoracic segment involvement.Fifteen patients had longitudinally extensive myelitic abnormalities (≥3 vertebral segments long).Seven enhancement patterns were encountered.Lesions were displayed in the spinal cord and brain in eight patients.Central gray matter involvement in the spinal cord was found in all the 19 patients.Conclusions Autoimmune GFAP astrocytopathy more frequently presents in females than in males.MRI of the spinal cord has complex presentations and longitudinally extensive myelitic abnormalities usually.Patients often show central gray matter involvement in the spinal cord.Myelitic abnormalities present more often in thoracic segment than in cervical segment.Abnormalities in lumbar segment are less encountered.

OBJECTIVETo explore the therapeutic efficiency of human telomerase reverse transcriptase promoter (hTERTp) mediated horseradish peroxidase (HRP) catalyzed effects of indole-3-acetic (IAA) on laryngeal squamous cell carcinoma with different radiosensitivity in vivo.

METHODSHuman laryngeal squamous cell carcinoma Hep-2 and Hep-2R cells were transplanted into nude mice. After growing to about 30 approximately 50 mm3, the tumor-bearing mice were randomly divided into eight groups: Hep-2 line: combined group (A), gene group (B), radiation group (C) and blank group (D); Hep-2R line: combined group (AR), gene group (BR), radiation group (CR) and blank group (DR). The phTERTp-HRP was delivered by intratumoral injection and the IAA by intraperitoneal injection, combined with 2 Gy daily radiation to a total dose of 30 Gy. The tumor volume was recorded. The cell apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay. The expression of HRP protein was detected by AP immunohistochemisty.

RESULTSThe tumor growth of combined groups was attenuated significantly and the tumor volume of Hep-2R blank group was the largest. The inhibition rate of each group was: A: 54.8%, B: 10.0%, C: 31.9%; AR: 52.7%, BR: 24.8%, CR: 17.0%. In the combined groups, necrosis and apoptosis of tumor cells were observed under the light microscope and the apoptotic index [A (16.6 +/- 1.3)% vs. AR (17.6 +/- 1.3)%] of tumor cells was highest (P < 0.05). The HRP protein expression of BR (33.3 +/- 8.9)% was higher than that of B (21.9 +/- 5.7)%, which was directly up-regulated in the tumors (45.0% vs. 54.8%, P < 0.05) after radiation.

CONCLUSIONIn the Hep-2- and Hep-2R-transplantation tumors in nude mice, hTERTp can be induced by radiation and enhance the expression of horseradish peroxidase (HRP) gene according to telomerase activity. hTERTp-HRP/IAA system, which has synergistic effects with radiation and inhibits the tumor growth by induction of apoptosis and necrosis, may be a new gene-radiation strategy for the treatment of laryngeal carcinoma.

Animals ; Apoptosis ; Cell Line, Tumor ; Combined Modality Therapy ; Female ; Genetic Therapy ; methods ; Horseradish Peroxidase ; genetics ; metabolism ; Humans ; Indoleacetic Acids ; metabolism ; pharmacology ; Laryngeal Neoplasms ; metabolism ; pathology ; therapy ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Promoter Regions, Genetic ; Radiation Tolerance ; Radiotherapy ; methods ; Random Allocation ; Telomerase ; genetics ; Tumor Burden

OBJECTIVETo evaluate the sedative and hypnotic effects of Foshouningshen decoction (FSNSD) and study its effects on expressions of 5-hydroxy tryptamine (5-HT) and 5-HT1A receptor (5-HTR) in the hippocampus in a rat model of insomnia.

METHODSMale KM mice were divided into control group, estazolam (0.4 mg/kg daily) group, and low-, moderate-, and high-dose FSNSD groups (daily dose of 12, 24, and 48 g/kg, respectively). After corresponding treatments for 1 week, the mice underwent sleep-inducing test with subthreshold and threshold doses of sodium pentobarbital. Forty-eight male SD rats were randomized into control group, insomnia model group, estazolam group (0.2 mg/kg daily), and low-, moderate-, and high-dose FSNSD groups (with daily dose of 6, 12, and 24 g/kg, respectively). Rat models of insomnia were established by intraperitoneal injection of 4-cholro-dl-phenylalanine (PCPA) at the daily dose of 350 mg/kg for 3 days, after which the rats received corresponding treatments via gavage for 1 week. The performance of the rats in open field test was recorded and the hippocampal expression of 5-HT was detected using ELISA; the expressions of 5-HTR protein and mRNA in the hippocampus were detected using immunohistochemistry and real-time PCR, respectively.

RESULTSIn the sleep-inducing test with a subthreshold dose of sodium pentobarbital, the mice treated with high-dose FSNSD showed a significantly higher rate of sleep onset than the control mice (P<0.05); in the test with a threshold dose of sodium pentobarbital, treatment with moderate- and high-dose FSNSD resulted in significantly prolonged sleeping time (P<0.01) and shortened sleep latency (P<0.05) in the mice. The rats in insomnia model group showed increased total distance in open field test (P<0.05) with significantly decreased content of 5-HT (P<0.01) and expressions of 5-HTR protein and mRNA in the hippocampus (P<0.01). Treatment of the rats with estazolam or high-dose FSNSD obviously decreased the total distance in open field test (P<0.05) and increased the content of 5-HT (P<0.05) and expressions of 5-HTR (P<0.01) in the hippocampus of rats with insomnia.

CONCLUSIONFSNSD can produce therapeutic effects on insomnia possibly by increasing 5-HT content and expressions of 5-HTR in the hippocampus.