AIM To establish an HPLC method for determining the contents of three constituents in Polygoni multiflori Radix Praeparata in plasma of atherosclerosis rats.METHODS After the rats were intragastrically administrated with Polygoni multiflori Radix Praeparata CMC-Na solution,the plasma was collected.The HPLC analysis was carried out on a 30 ℃ thermostatic Hypersil C1s column (250 mm ×4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-0.03％ phosphoric acid flowing at 0.9 mL/min in a gradient elution manner,and the detection wavelength was set at 280 nm.DAS2.0 software was applied to drawing concentration-time curves and calculating pharmacokinetic parameters.RESULTS Stilbene glucoside,emodin and physcion showed good linear relationships within the ranges of 61.25-6 125 μg/L (r =0.999 8),12.6-3 150 μg/L (r =0.999 3) and 24.1-6 030 μg/L (r =0.999 5),respectively.The method recoveries were 99.5％-105.8％ with the RSDs of 1.3％-3.3％,while the extraction recoveries were 87.2％-96.3％ with the RSDs of 3.2％-5.9％.The pharmacokinetic behaviors of three constituents all accorded with two-compartment model,but their contents in plasma were much lower than those in medicinal material.CONCLUSION The bioavailabilities of stilbene glucoside,emodin and physcion are relatively low in plasma of atherosclerosis rats,which may be related to constituents' intestinal absorption after intragastric administration with Polygoni multiflori Radix Praeparata.

Animals ; Cell Proliferation ; Hepatic Stellate Cells ; Liver Cirrhosis/metabolism* ; Protein Tyrosine Phosphatase, Non-Receptor Type 6 ; Rats

Objective?To observe the clinical effect of FURS combined with ultrasound guided all seeing needle treatment of low pole renal calculi.?Methods?From January 2016 to January 2017, 63 patients with low pole renal calculi were enrolled in this study. 16 patients with multiple stones and 47 cases with single stones were treated by FURS combined with ultrasound guided all seeing needle. We retrospectively analyzed all the patients clinical data, including the operation time, bleeding volume, stone clearance rate, the incidence of postoperative complications, then assess the safety and effectiveness of the surgical approach.?Results?The operation time was 30 ~ 60 min, the average time was 45 min, postoperative complications in 3 cases, 2 cases of pain; no blood transfusion; 2 cases of percutaneous nephrolithotomy. Postoperative hospital stay wad 2 ~ 3 d, an average of 2 d, postoperative residual stone in 3 cases, 2 weeks after the treatment of external lithotripsy, 1 case of stone clearance rate of 100.0%.?Conclusion?Retrograde FURS combined with ultrasound guided visual puncture for renal calculi is safe, effective and feasible.

BACKGROUNDOssification of the ligamentum flavum (OLF) is being increasingly recognized as a cause of thoracic myelopathy. This study was to describe a rare clinical entity of spinal cord kinking (SK) in thoracic myelopathy secondary to OLF.

METHODSThe data of 95 patients with thoracic myelopathy secondary to OLF were analyzed retrospectively. The incidence and location of SK were determined using preoperative magnetic resonance imaging (MRI). The clinical presentation and radiological characteristics in patients with SK were analyzed. Posterior en bloc laminectomy with OLF was performed, and the surgical results were evaluated.

RESULTSSK was found in seven patients (7.4%) based on preoperative MRI. The patients included one male and six females with an average age of 55.6 years (range, 48-64 years). Five patients presented with radiculomyelopathy and two presented with typical thoracic myelopathy of spastic paraparesis. In all cases, the kinking was located just above the end of the spinal cord where the conus medullaris (CM) was compressed by the OLF. The degree of SK varied from mild to severe. The tip of the CM was located between the upper third of T11 to the lower third of L1, above the lower edge of L1. With an average follow-up of 30.4 months, the modified Japanese Orthopedic Association score significantly improved from 5.7 ± 1.8 preoperatively to 8.9 ± 1.4 postoperatively (t = 12.05; P < 0.0001) with an improvement rate of 63.1 ± 12.3%.

CONCLUSIONSSK is a rare radiological phenomenon. It is typically located at the thoracolumbar junction, where the CM is compressed by the OLF. Our findings indicate that these patients may benefit from a posterior decompressive procedure.

Female ; Humans ; Ligamentum Flavum ; pathology ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Ossification, Heterotopic ; complications ; Radiography ; Spinal Cord Compression ; diagnosis ; diagnostic imaging ; surgery ; Spinal Cord Diseases ; diagnosis ; diagnostic imaging ; etiology ; surgery

OBJECTIVETo investigate the roles of Kruppel-like factor 6 (KLF6) and its splice variant KLF6V on suppressing growth and inducing differentiation of human hepatocellular carcinoma hepG2 cells.

METHODKLF6V cDNA was amplificated by RT-PCR from human hepatocellular carcinoma (HCC) tissue and then sequenced. The recombinant vectors expressing KLF6 variant (KLF6V) were constructed using molecular clone technology based on established plasmid pcDNA3.1A(-)/wtKLF6. KLF6V or KLF6-transfected HepG2 cells were established after being screened with G418. Growth activity of HepG2/KLF6 or HepG2/KLF6V cells was detected by in vitro MTT assay. Expression of p21WAF1 or cyclin D1 protein was detected by Western blot, and expressions of AFP or ALB protein were measured by radioimmunoassay.

RESULTSA novel alternatively spliced transcript of the human KLF6 gene was found and its sequencing revealed that the variant form of KLF6 lacked 126nt and its encoded protein products had a deletion of 42 aa near the COOH-terminal amino acid in comparison with full-length KLF6. Although KLF6 alternative splicing was present in both normal and cancerous tissues, expression of the KLF6 splice variants seemed to be up-regulated in HCCs tissues. The isoform of KLF6 proteins antagonized the ability of wild-type KLF6 to up-regulate p21 expression or down-regulate cyclin D1 expression and suppress HepG2 cell proliferation. KLF6 gene increased albumin production and decreased alpha fetoprotein production of the cells.

CONCLUSIONThe isoform of KLF6 protein, present in HCC tissue, antagonizes the ability of wild-type KLF6 to suppress cell proliferation and induce cellular differentiation.

Cell Differentiation ; Cell Proliferation ; DNA, Complementary ; Gene Expression Regulation, Neoplastic ; Hep G2 Cells ; Humans ; Kruppel-Like Factor 6 ; Kruppel-Like Transcription Factors ; genetics ; Protein Isoforms ; genetics ; Proto-Oncogene Proteins ; genetics ; Transfection

OBJECTIVETo investigate two single nucleotide polymorphism sites of the promoter region in T cells immunoglobulin domain and mucin domain protein-3 (TIM-3) and detect their relationship with allergic asthma in a population of adult Hans from Hubei province of China.

METHODSThe polymorphisms were detected with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and allelic specific polymerase chain reaction (ASPCR). The genotype and allele frequencies were calculated and analyzed.

RESULTSThe genotype frequencies of CC, CT and TT in -1541 C/T polymorphism were 0.961, 0.039 and 0 respectively in the healthy population and were 0.935, 0.065 and 0 respectively in the allergic asthma population. No significant difference in genotype and alleles frequencies was found between the allergic asthma patients and control subjects (P=0.314, P=0.321). The genotype frequencies of GG, GT and TT in -574 T/G polymorphism were 0.992, 0.008 and 0 respectively in the healthy population and were 0.941, 0.059 and 0 respectively in the allergic asthma population. There was significant difference in genotype and allele frequencies between the allergic asthma patients and control subjects (P=0.046, P=0.048).

CONCLUSIONThere are polymorphism sites of the promoter region in TIM-3 , and one of these sites, the -574 G/T polymorphism site, may be associated with allergic asthma in the population of adult Hans from Hubei province of China.

Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; Asthma ; genetics ; China ; ethnology ; Female ; Hepatitis A Virus Cellular Receptor 2 ; Humans ; Male ; Membrane Proteins ; Middle Aged ; Polymorphism, Genetic ; Population ; Receptors, Virus ; genetics

OBJECTIVETo find some effective short interfering RNA's sites targeting HBV surface gene sequence using shRNA expression vectors.

METHODSFour shRNA expression vectors targeting HBV surface gene sequence were constructed based on pAVU6 + 27 vector, and cotransfected into AD293 cells with HBs-EGFP fusion gene plasmid. The changes of HBs-EGFP image were detected by FACS and microscopy. The HBs-EGFP mRNA expression was evaluated by RT-PCR.

RESULTSFour shRNA expression vectors and HBs-EGFP fusion gene plasmid were successfully constructed. pAVU6 + 4sh579 vector inhibited the HBs-EGFP expression by 69.8% in AD293 and suppressed the HBs-EGFP mRNA expression by 74.6%.

CONCLUSIONSThe results showed that the 579 site of HBV surface gene sequence was an effective target and pAVU6 + 4sh579 vector could suppress the HBs-EGFP expression in AD293 cells

Gene Expression Regulation, Viral ; Gene Silencing ; Gene Targeting ; methods ; Hepatitis B Surface Antigens ; genetics ; metabolism ; Humans ; RNA Interference ; RNA, Small Interfering ; genetics ; RNA-Induced Silencing Complex ; genetics

OBJECTIVETo investigate the efficacy of 2-month course of sleeping position correction in the treatment of positional plagiocephaly in infants aged <8 months.

METHODSA total of 73 infants with positional plagiocephaly between January 2015 and June 2016 were divided into treatment group (n=46) and control group (n=27) according to parents' wishes. The treatment group received sleeping position correction, while the control group received sleep curve mattress. The oblique diameters A and B in the two groups were measured and the cranial vault asymmetry (CVA) was calculated before and after treatment. The severity of positional plagiocephaly based on CVA was compared between the two groups before and after treatment. The Gesell Developmental Scale was used to determine the developmental quotients (DQs) in the motor, adaptive, language, and social domains in the two groups before and after treatment.

RESULTSBefore treatment, there were no significant differences in oblique diameters A and B, CVA, and DQs in the four specific domains between the two groups (P>0.05). After 2 months of treatment, the treatment group had a significantly greater oblique diameter B and a significantly smaller CVA than the control group (P<0.05); there were no significant differences in DQs in the four specific domains between the two groups (P>0.05). After treatment, both groups had significant improvements in oblique diameters A and B, CVA, and DQs in the motor and adaptive domains (P<0.01); moreover, the treatment group showed a significant improvement in the DQs in the social domain (P<0.01). There was no significant difference in the severity of positional plagiocephaly between the two groups before and after treatment (P>0.05).

CONCLUSIONSFor infants with positional plagiocephaly, sleeping position correction has better efficacy and is more convenient and economical than the sleep curve mattress, so it holds promise for clinical application.

Female ; Humans ; Infant ; Male ; Plagiocephaly, Nonsynostotic ; etiology ; therapy ; Posture ; Severity of Illness Index ; Sleep

Objective To optimize the extraction conditions of Schisandra sphenanthera(SS)seed oil by supercritical CO2ex?traction and identify its components by GC-MS.Methods SS seed oil was used as tested material,response surface methodology was used to optimize the process of supercritical CO2extraction,and GS-MS method was used to analyse SS seed oil composition.Results A model of an equation was established,which could be used to optimize the process parameters of supercritical CO2extraction of SS seed oil. The optimum extraction parameters were as follows:the extraction pressure was 33 MPa,the extraction temperature was 53℃,the extraction time was 90 min,the CO2flow rate was 21.40 ml/min.In this condition,the extraction rate of SS seed oil was 7.97%.The SS seed oil was analyzed by GC-MS,and 23 compounds were identified.In these compounds,(1α,4a.β,8a.α)-1,2,3, 4,4a,5,6,8a-octahydro-7-methyl-4-methylene-1-(1-methylethenyl)-naphthalene,(-)-1,7-dimethyl-7-(4-methyl-3-pentenyl)-tricyc?lo[2.2.1.0(2,6)]heptane and(R)-2,4a,5,6,7,8-octahydro-3,5,5,9-tetramethyl-1H-(σ-phenyl)cycloheptene had the content of more than 10% and the contents were 27.78%,14.77% and 13.12% respectively.Conclusion This process has high extraction rate, fast speed and simple operation,and can be used for the extraction of SS seed oil.

Objective To analyze the possibilities of screening SPF rabbits and guinea pigs from conventional animals, viral antibodies of the conventional rabbits and guinea pigs bred by licensed companies in Guangdong province during 2014-2016 were determined according to the standard of SPF animals in GB14922. 2. Methods Nine batches of 167 rabbit sera and 155 guinea pig sera were sampled from 6 companies. Serum antibodies to virus were determined by ELISA according to GB14922. 2. Results Positivity of antibody to rabbit hemorrhagic disease virus (RHDV) was 82. 2%(129/157) for the vaccinated rabbits, and negative result were obtained for unvaccinated rabbits. Positive rate of rabbit rotavirus (RRV) was 42. 5% (71/167). No positive antibody responses to Sendai virus were detected out in all rabbits. The positive rates of guinea pig reovirus type III (REO-3) and pneumonia virus of mice (PVM) were 52. 9%(82/155)and 20% (31/155) respectively. Antibody responses to Sendai virus ( SV) and lymphocytic choriomeningitis virus ( LCMV) were negative in all guinea pigs. Conclusions Although the conventional rabbits and guinea pigs could meet the national standard, higher infection rates of virus excluded that SPF animals emerged in conventional animals, indicating that selection of SPF animals from conventional colonies is impracticable.