Objective To investigate the anti-rotavirus effect of Gegen Qinlian decoction on rotavirus (RV)-inducing diarrhea neonatal mice model, and provide evidences for pharmacological and pharmacodynamic studies. Methods Neonatal diarrhea mice model of Kunming family infected with RV-Wa was developed. Symptom, diarrhea ratio and severity level, histological changes of neonatal mice’s small intestine and lung was assessed after Gegen Qinlian decoction (0.78 g/kg) treatment. Results Diarrhea ratio, diarrhea severity level, histological changes of neonatal mice’s small intestine and lung were obviously decreased after Gegen Qinlian decoction (0.78 g/kg) treatment. Conclusion Gegen Qinlian decoction has anti-rotavirus effect on RV induced diarrhea.

Objective To observe the difference of vascular endothelial growth factor (VEGF) and myeloperoxidase (MPO) between diabetic and non-diabetic patients with maintenance hemodialysis (MHD),and to explore whether it was associated with duration of hemodialysis.Methods A total of 120 patients with MHD were divided into diabetic group (40 cases) and non-diabetic group (80 cases) according to the primary disease.The blood samples from the patients before dialysis were selected to test the serum VEGF and plasma MPO and other indicators.The blood samples from April 2012 to March 2013 were labeled diabetic group 1(40 cases) and non-diabetic group 1 (80 cases).The blood samples from April 2013 to March 2014 were labeled diabetic group 2 (40 cases) and non-diabetic group 2 (80 cases).Results The serum VEGF and plasma MPO levels were (74.63 ± 47.43) ng/L,(300.63 ± 235.37) μ g/L in diabetic group 1,and (63.69 ± 43.23) ng/L,(275.35 ± 216.32) μ g/L in non-diabetic group 1,and there were significant differences between two groups (P ＜ 0.05).The serum VEGF and plasma MPO levels were (83.32 ± 40.38) ng/L,(414.12 ±265.52) μg/L in diabetic group 2,and (70.89 ±39.74) rig/L,(289.45 ±202.85) μg/L in non-diabetic group 2,and there were significant differences between two groups (P ＜ 0.05).The correlation analysis showed that VEGF was positively correlated with MPO in diabetic group 1 and diabetic group 2 (r =0.632 and 0.763,P ＜ 0.05),and VEGF was positively correlated with MPO in non-diabetic group 1 and non-diabetic group 2 (r =0.610 and 0.713,P ＜ 0.05).Conclusions Compared with those in non-diabetic MHD patients,VEGF and MPO levels are significandy higher in diabetic MHD patients.In non-diabetic MHD patients and diabetic MHD patients,VEGF and MPO levels will rise gradually with duration of hemodialysis.The expressions of VEGF and MPO are associated with each other.

Four small GTPase genes which may be relative to sclerotial development were firstly cloned from medicinal fungus Polyporus umbellatus using rapid amplification of cDNA end PCR (RACE) method. The results showed that full-length cDNA of PuRhoA was 698 bp contained 585 bp ORF, which was predicted to encode a 194 amino acid protein with a molecular weight of 21.75 kD with an isoelectric point (pI) of 6.44; the full length cDNA of PuRhoA2 was 837 bp in length and encoded a 194 amino acid protein with a molecular weight of 21.75 kD and an isoelectric point (pI) of 6.33; the full length cDNA of Puypt1 was 896 bp in length and encoded a 204-aa protein with a molecular weight of 22.556 kD and an isoelectric point (pI) of 5.75; the full length cDNA of PuRas was 803 bp in length and encoded a 212-aa protein with a molecular weight of 23.821 kD and an isoelectric point (pI) of 5.2. There are fani acyl transferase enzyme catalytic site and myrcene-transferase enzyme catalytic site in PuRhoA1 while the PuRhoA2 only possess myrcene-transferase enzyme catalytic site. Puypt1 contains the Rab1-Ypt1 conserved domain of small GTPase family and PuRas contains the fani acyl transferase enzyme catalytic site. According to the phylogenetic analysis all these four small GTPase clustered with basidiomycete group. Quantitative real-time PCR analysis revealed that Puypt1, PuRas and PuRhoA1 transcripts were significantly higher in the beginning of sclerotial formation than that in the mycelia, whereas the transcripts levels of PuRhoA2 gene were particularly lower in sclerotia than that in mycelia, suggesting that these four genes might be involved in P umbellatus selerotial development.

Objective To compare the clinical efficacy of intralesional betamethasone versus triamcinolone acetonide acetate in the treatment of active alopecia areata. Methods A total of 160 patients with active alopecia areata were divided into two groups, test group (n = 100) treated with intralesional betamethasone, and control group (n = 60) treated with intralesional triamcinolone acetonide. Both injections were given once every 3 weeks for 12 consecutive weeks. Results After 12-week treatment, the cure rate, response rate, and total response rate were 60.0%, 32.0% and 92.0% in the test group, respectively, compared to 41.7%, 31.67% and 73.3% in the control group, respectively. A significant increase was observed in the cure rate and response rate in the test group compared with the control group (χ2 = 10.25, 5.06, P < 0.01 and 0.05). During the treatment course, 8 (8%) patients in the test group and 9 (15%) patients in the control group developed localized atrophy of the scalp; 8 (8%) patients in the test group and 3 (5%) patients in the control group developed localized folliculitis; no significant difference was observed between the two groups in the occurrence of adverse reactions (P> 0.05). Conclusion Intralesional use of compound betamethasone injection has a notable therapeutic effect on alopecia areata.

Objective To evaluate the effect of chlamydiaphage virus protein 2(Vp2) on the recombinant virus and virus screening research, and it clinical value thereof. Methods To compare the Vp2 protein sequences to get the conserva-tive region with COBALT. A phylogenetic tree was built with ProteinBlast of Distance tree. The amino acid sequence in the high conservative region was predicted by the methods of Gamier-Robson and Chou-Fasman, and its flexibe regions were predicted by Karplus method. The hydrophilicity plot was predicted by Kyte-Doolittle and Hopp-Woods method. The sur-face probability was analysed by Emini, and the antigenic index was analysed by Jameson-Wolf method. Results The six Chlamydiaphage Vp2 proteins were the highly conserved sequences. There were obvious differences between Chp1Vp2 and other 5 Vp2 proteins. There were the main structure-alpha helix and some cell epitopes in the high conserved region. Con-clusion Vp2 protein is the important component of chlamydia phage capsid with the conservative nature. Vp2 protein has complicated structures and high conservative region with strong immunogenicity, playing a practical value of research in vi-rus recombinantment and screening the wild strains of chlaymdia trachomatis phage.

We wished to assess the role of chlamydia micro virus capsid protein Vp3 in recombinant molecules, chart its molecular evolution, screen the wild-type strain, and reveal its value in clinical research. Using a protein BLAST multiple-alignment program, we compared various strains of Chlamydia micro virus capsid protein Vp3 sequences. Using a "distance tree" of those results, we created a phylogenetic tree. We applied the Karplus-Schulz method of flexible-region analyses for highly conserved alignments of amino-acid sequences. Gamier-Robson and Chou-Fasman methods were employed to analyze two-level structures of sequences. The Emini method was used for analyses of the accessibility of surface epitopes. Studies of hydrophilic proteins were undertaken using Kyte-Doolittle and Hopp-Woods methods. Analyses of antigen epitopes helped to reveal the antigen index using the Jameson-Wolf method. All sequences in the six strains of chlamydia micro virus capsid protein Vp3 were highly conserved, with the main differences being between Vp3 protein in Chp1 and the other five strains of the micro virus. The viral strain of Vp3 protein was based mainly on micro-alpha helix structures, and multiple epitopes were noted in highly conserved regions. Vp3 protein was highly conserved structurally, and was an important protein of the chlamydiaphage capsid. Vp3 protein has a complicated molecular structure, highly conserved regions with strong immunogenicity, and has considerable research value.
Amino Acid Sequence ; Capsid Proteins ; chemistry ; genetics ; immunology ; Chlamydia ; genetics ; immunology ; Conserved Sequence ; Epitope Mapping ; Evolution, Molecular ; Molecular Sequence Data ; Recombination, Genetic

Objective To observe secreted protein acidic and rich in cysteine (SPARC) levels in serum and foot muscle tissue of patients with type 2 diabetic foot (DF).Methods All participants were divided into four groups with 40 cases in each group, including type 2 diabetic patients without (DMA) and with (DMB) lower limb arterial sclerosis and peripheral neuropathy, DF group, and normal control (NC) group.The muscle tissues of foot from DF group (n =6) and NC group (n =6) were taken.Serum SPARC level was measured with ELISA.RT-PCR, Western blot, immunofluorescence, and immunohistochemistry were used to examine SPARC mRNA and protein expressions in the foot muscle.Results Serum SPARC levels were higher in DMA and DMB groups compared with NC and DF groups[(1 040.48 ±212.12 and 1 068.36 ± 165.45 vs 841.93 ± 144.57 and 835.43 ± 188.37) ng/L, P＜ 0.01].There was no significant difference in serum SPARC level between DF and NC groups or DMA and DMB groups (P＞0.05).The expression levels of SPARC mRNA and protein in the foot muscle were higher in DF compared with NC group (P＜0.05).Conclusion SPARC mRNA and protein expressions in foot muscle tissue are higher in DF group compared with control group, indicating that SPARC may participate in the repair and healing of damaged muscle tissue of diabetic foot.

Objective To quantitatively analyze the value of color Doppler twinkling artifact and CT in distinguishing urinary stone composition of uric acid from that of cystine,and to assess the ability of color Doppler twinkling artifact and CT to diagnose urinary stone composition of uric acid.Methods Artificial stones made of uric acid and cystihe were inserted in porcine kidneys respectively.Color Doppler ultrasound and CT scanning were performed in vitro and twinkling artifact videos were recorded.The CT attenuation value,the width and length of twinkling artifact (TAW,TAL) were measured.The color pixels representing twinkling artifact intensity (TAI) were calculated.The differences between artificial cystine and artificial uric acid stones in TAW,TAL,TAI and CT attenuation value were compared.The receiver operating characteristic (ROC) curve was used to assess the ability of TAW,TAL,TAI and CT attenuation value to diagnose urinary stone composition of uric acid.Results The TAW [(4.16 ± 1.54)mm vs (4.97 ± 1.93)mm],TAL[(3.75 ± 1.05)mm vs (5.12 ± 2.51)mm] and TAI [(3 005.50 ± 812.33) vs (5 433.86±2 505.81)] of artificial cystine stones were less than those of artificial uric acid stones while the CT attenuation value of artificial cystine stones [(573.75 ± 110.10)HU vs (364.09 ± 67.28)HU] were greater (P ＜0.05).The ROC curve was displayed to diagnose urinary stone composition of uric acid,and the area under curve of CT attenuation value was bigger than that of TAI (0.936 vs 0.817,Z =2.308,P =0.021).Combination of color Doppler twinkling artifact and CT value would dramatically increase the diagnostic accuracy to 98％.Conclusions Both color Doppler ultrasound and CT were reliable examinations for distinguishing urinary stone composition of cystine from that of uric acid.Combination of color Doppler ultrasound and CT would be good references to diagnose urinary stone composition of uric acid.

Objective To investigate the diagnostic value of twinkling artifact in urinary calcium stones.Methods Calcium oxalate monohydrate,hydroxyapatite and whitlockite stone phantoms were prepared and embedded in porcine kidneys for ultrasound scanning.The length and width of twinkling artifact were measured.The intensity of twinkling artifact and the contrast-to-noise ratio (C/N ratio) of acoustic shadowing were recorded and all the data was analyzed statistically.Results All the stone phantoms generated twinkling artifact and acoustic shadowing.The difference of the intensity,length and width of twinkling artifact and the C/N ratio of acoustic shadowing had statistically significant each composition (P ＜0.01).There was no overlap between 95％ confidence interval of the intensity of twinkling artifact for any two compositions,and any two compositions of stones could be differentiated by the intensity of twinkling artifact (P ＜0.05).The C/N ratio of acoustic shadowing of calcium oxalate monohydrate and whitlockite stone phantoms were higher than those of hydroxyapatite ones (P ＜0.05).The receiver operating characteristic (ROC) curve displayed that the cutoff value of 1 127 of the intensity of twinkling artifact could be used to diagnose hydroxyapatite stones.The sensitivity,specificity,positive predictive value (PPV),negative predictive value (NPV) and area under the curve (AUC) were 74 ％,68 ％,69.8％,72.3％ and 0.743 respectively.Conclusions Twinkling artifact could be used to differentiate the calcium stones,which will be helpful for clinical treatment.

Objective To investigate the serum concentrations of secreted protein acidic and rich in cysteine (SPARC) in patients with diabetic nephropathy and SPARC mRNA and protein expressions in renal tissue of db/db mice.Methods Serum SPARC levels in normal subjects and patients with type 2 diabetes mellitus (without diabetic nephropathy),chronic renal failure (without diabetes mellitus),and diabetic nephropathy were determined with enzyme-linked immunosorbent assay.RT-PCR,Western blot,and immunofluorescence were used to detect the mRNA and protein expressions of SPARC in renal tissue of db/db mice.Results The serum level of SPARC in diabetic nephropathy group was significantly higher than those in normal group,type 2 diabetes mellitus,and chronic renal failure group [(5.78 ± 1.41 vs 3.58 ±0.41,4.51 ± 1.08,and 3.81 ± 1.16) μg/L,P＜0.05 or P＜0.01].The SPARC level in the type 2 diabetes mellitus group was higher than that in normal group (P＜0.05),but there was no difference between normal group and chronic renal failure.SPARC mRNA and protein levels in renal tissue of db/db mice were higher compared with the littermates (P ＜ 0.05).Conclusions The long term hyperglycemic state in patients with diabetic nephropathy causes pathological change of renal tissue.Simultaneously,increased secretion of SPARC from renal tissue results in elevation of serum SPARC level and may play a role in pathological change of diabetic nephropathy.