Objective To describe the natural history of cervical intraepithelial neoplasia(CIN)Ⅰand the biologic factors associated with the progression of CINⅠ and to analyze the predictive values of p16INK4a protein for the progression of CINⅠ. Methods From August 2010 to July 2013, 104 patients referred for abnormal cytology [≤ low-grade squamous intraepithelial lesion (LSIL); including negative for intraepithelial lesion or malignancy (NILM), atypical squamous cells of undetermined significance (ASCUS), LSIL] and high-risk (HR) HPV positive,and were diagnosed CINⅠ by colposcopy-assisted biopsy and followed at 1-year intervals in the First Affiliated Hospital of Nanjing Medical University. In order to analyze the relationship between the progression of CINⅠ with clinical biologic factors, including patient age, cervical cytology before colposcopy, loads of HR HPV, HPV L1 capsid protein, p16INK4a protein,χ2 tests was used to compare the different frequencies of factors in groups of progressed and persisted/regressed CINⅠ, then five factors with progressed CINⅠwere processed into binary logistic regression analysis. Results (1) In the first year of follow-up, among 104 patients(including 15 cases NILM,78 cases ASCUS,11 cases LSIL), 52 cases of them were NILM and HR HPV negative, 30 cases were negative for intraepithelial lesion, 10 cases were CINⅠ, 5 cases were CINⅡand 7 cases were CINⅢ. In total, 82 cases (78.8%,82/104) cases had regressed, 10 cases (9.6%,10/104) persisted, 12 cases (11.5%,12/104) progressed [including 5 cases (4.8%,5/104) progressed to CIN Ⅱ, 7 cases (6.7%,7/104) progressed to CIN Ⅲ, none progressed to invasive cancer]. (2) All patients, 88 cases of them accepted immunohistochemical detection the expression of p16INK4a protein. The result shown that 30 cases (34%,30/88) were positive and 58 cases (66%,58/88) were negative. And 94 cases accepted immunocytochemical detection the expression of HPV L1 capsid protein, 49 cases (52%,49/94) were positive and 45 cases (48%,45/94) were negative. (3) Univariate analysis showed that age of the patient, loads of HR HPV, cervical cytology before colposcopy and the expression of HPV L1 capsid protein were not risk factors of the progression of CINⅠ(all P>0.05) except for the expression of p16INK4a protein (P<0.05). Multivariable analysis found that p16INK4a protein positive was associated with progression of CINⅠ(OR=5.1,95%CI:1.162-22.387,P=0.031). (4) Thirty-one cases were p16INK4a protein positive, 8 cases (27%,8/30) of them progressed,while 4 cases (7%,4/58) of 58 cases with p16INK4a protein negative progressed,in which there were significant difference (P<0.05). The sensitivity was 75%, the specificity was 71%, the positive predictive value was 27%and the negative predictive value was 93%for progression to CINⅡ-Ⅲof p16INK4a protein staining. Conclusions The progression rate of CINⅠwith abnormal cytology (≤LSIL) and HR HPV positive was lower, and there was no progression to invasion at 1-year intervals. Immunostaining of p16INK4a protein as the risk factors of CINⅠprogression could have a role in prediction of CINⅠand the management of high-risk CINⅠ.

Objective To investigate the CT characteristics of anaplastic thyroid carcinoma and evaluate the diagnostic value of CT in this disease.Methods The CT findings of 10 patients with pathologically proved anaplastic thyroid carcinoma were retrospectively reviewed.The patients included 7 females and 3 males.Their age ranged from 25.0 to 78 years with median of 61 years.Multi-slices plain and post contrast CT scans were performed in all patients.Results Unilateral thyroid was involved in 6 patients.Unilateral thyroid and thyroid isthmus were both involved in 2 patients due to big size.Bilateral thyroid were involved in 2 patients.The maximum diameter of anaplastic thyroid carcinoma ranged from 2.9-12.8 cm with mean of (4.5 ± 1.4) cm.All lesions demonstrated unclear margins and envelope invasion.The densities of all lesions were heterogeneous and obvious necrosis areas were noted on precontrast images.Seven lesions showed varied calcifications,and coarse granular calcifications were found in 5 lesions among them.All lesions showed remarkable heterogenous enhancement on post-contrast CT.The CT value of solid portion of the tumor increased 40 HU after contrast media administration.The ratios of CT value which comparing of the tumor with contralateral sternocleidomastoid muscle were 0.69-0.82 (0.76 ± 0.18)and 1.25-1.41 (1.33 ± 0.28)on pre and post CT,respectively.Enlarged cervical lymph nodes were found in 6 cases (60.0％).It showed obvious homogeneous enhancement or irregular ring-like enhancement on post-contrast images and dot calcifications were seen in 1 case.Conclusions Relative larger single thyroid masses with coarse granular calcifications,necrosis,envelope invasion,remarkable heterogeneous enhancing and enlarged lymph nodes on CT are suggestive of anaplastic thyroid carcinoma.

Arginine Deiminase(ADI) was purified to homogeneity using ammonium sulfate precipitation,Q-Sepharose Fast Flow anion exchange chromatography and SephadexG-75 gel filtration chromatography. This purification protocol resulted in a 34.5-fold purification of ADI with 31.4% final yield. A molecular weight of about 190 kD determined by native gradient polyacrylamide gel electrophoresis. The enzyme has only one kind of 46 kD subunit determined by SDS-PAGE. Combining the results from the two kinds of electrophoresis,the authors deduce that the enzyme may be a tetramer. The optimum pH and temperature for lipolytic activity of ADI was pH 6.5 and 50℃,respectively. It was extremely stable at 45℃ and retained 97.9% of its original activity for 30 min. The stability declined rapidly as soon as the temperature rose over 50℃. ADI was highly stable in the pH range from pH 5-8. ADI acted on L-arginine but not on D-arginine. ADI catabolism was dependent on metal ions. At their adequate concentration,Mn2+,Mg2+ and Co2+ were the effective promoter,while superfluous Zn2+and Co2+ inhibited ADI activity. L-citrulline did not act on ADI,but L-ornithine inhibited ADI activity. The degradation of L-arginine with ADI catalysis was according to simple Michaelis-Menten equation. The Michaelis constant was 3.2686 mmol/L and the maxi-mum velocity was 2.44 ?mol/min.

Background:Conventional treatment has limited efficacy in relapsed/refractory B-cell lymphoma.Since chimeric antigen receptor Tcell (CAR-T) technology has shown high safety and results in high remission rates,we investigated its efficacy and safety in B-cell lymphoma treatment and analyzed potential affecting factors to provide evidence for therapeutic strategies and applications.Methods:We searched databases including PubMed,Embase,and Cochrane up to July 2019.Meta-analysis 1 was conducted to study the efficacy of CAR-T cell for treating B-cell lymphoma,measuring the response rate and complete remission rate as outcomes.Sub-group analysis was performed for age,pathological type,target antigen,co-stimulatory molecule,and conditioning chemotherapy.Meta-analysis 2 was undertaken on the safety of the treatment with the incidence rate of toxicity (cytokine-releasing syndrome [CRS],neurotoxicity) as an outcome.Results:Seventeen studies were included in the systematic review and meta-analysis.It was found that CAR-T cells had good therapeutic effects in the following cases:B-cell lymphoma (patients ≥65 years old);diffuse large B-cell lymphoma pathological type;patients with treatment target antigen other than CD 19;patients treated with co-stimulatory molecules other than CD28,including 4-1BB+CD28 or 4-1BB;and patients treated with cyclophosphamide/fludarabine pre-treatment protocol conditioning chemotherapy.Although the CRS and neurotoxicity incidences were high,most were reversible with minimal risk of death.Conclusion:CAR-T cell treatment is safe for clinical annlication:however,toxicity effects should be monitored.

AIM: To establish the quality standards for Chinese extractum angelicae liquidum. METHODS: TLC was used to identify ferulic acid and ligustilide and HPLC to determine the content of ferulic acid and ligustilide.HPLC was performed on a Diamonsil ODS-C_18 analytical column(250 mm?4.6 mm,5 ?m) with gradient elution(0-15 min,38%A;15-20 min,38%A70%A;20-40 min,70%A;40-45 min,70%A-38%A) of methanol(A,containing 0.4% glacial acetic) and 0.4% glacial acetic acid(B) at the flow rate of 1.0 mL/min.The diode array detection wavelength was set at 323 nm and the column temperature was at 35 ℃. RESULTS: The linear range of ferulic acid and ligustilide were from 1.008 ?g/mL to 10.08 ?g/mL and 9.985 ?g/mL to 99.85 ?g/mL,the average recoveries of both were 98.11% and 101.61%,RSD were 1.58% and 1.32%.CONCLUSION: The method is rapid,simple and accurate with high reproducibility and can be used to control the quality of Chinese extractum angelicae liquidum.

AIM: To establish the quality standards for Chinese extractum angelicae liquidum. METHODS: TLC was used to identify ferulic acid and ligustilide and HPLC to determine the content of ferulic acid and ligustil-ide. HPLC was performed on a Diamonsil ODS-C_(18) analytical column(250 mm×4.6 mm, 5 μm) with gradient elu-tion(0-15 min, 38%A; 15-20 min, 38%A-70%A; 20-40 min, 70%A; 40-45 min, 70%A-38%A) of methanol (A, containing 0.4% glacial acetic) and 0.4% glacial acetic acid(B) at the flow rate of 1.0 mL/min. The diode array detection wavelength was set at 323 nm and the column temperature was at 35℃. RESULTS: The linear range of ferulic acid and ligustilide were from 1.008 μg/mL to 10.08 μg/mL and 9.985 μg/mL to 99.85 μg/mL,the average recoveries of both were 98.11% and 101.61%, RSD were 1.58% and 1.32%. CONCLUSION: The method is rapid, simple and accurate with high reproducibility and can be used to control the quality of Chinese extractum angelicae liquidum.

One unusual human G3P[3] group A rotavirus (RVA) strain M2-102 was identified in stool sample collected from a child with diarrhea in Guangxi Province, China in 2014. It is well known that G3P[3] is a genotype commonly identified in feline and canine RVAs. However, the preliminary phylogenetic analyses of the VP7 and VP4 genes of strain M2-102 indicated that these two genes were closely related to bat RVA strain MYAS33 and simian strain RRV, respectively, whereas both clustered distantly to feline/canine-like RVA strains. In this study, full genome sequencing and molecular analyses were conducted to obtain the true origin of strain M2-102. It was revealed that strain RVA/Human-wt/CHN/M2-102/2014/G3P[3] exhibited a G3-P[3]-I3-R3-C3-M3-A9-N3-T3-E3-H6 genotype constellation for VP7-VP4-VP6-VP1-VP2-VP3-NSP1-NSP2-NSP3-NSP4-NSP5 genes. Phylogenetic analyses revealed that 5 genes (VP7, VP1, VP2, NSP2 and NSP3) from strain M2-102 were closely related to those of bat strain MYAS33 from Yunnan Province which was thought a true bat RVA strain rather than a virus transmitted between species, while another 5 genes (VP4, VP3, NSP1, NSP4 and NSP5) clustered closely with those of simian strain RRV, yet the VP6 gene was closely related to that of human G3P[9] strain AU-1 and AU-1-like RVAs. The epidemiological data indicated that the child infected with M2-102 came from a countryside village, located in Dong Autonomous County of Sanjiang (subtropical hilly wooded area), Liuzhou city in Guangxi Province which might provide natural environment for reassortment events occurring among animal and human RVAs. Therefore, the data suggest that human strain M2-102 might originate from multiple reassortment events among bat, simian and human AU-1-like RVAs, yet it is not clear whether the genomic backbone based on bat MYAS33 (5 genes) and simian RRV (5 genes) like rotaviruses had been obtained through reassortment before being transmitted to the human. This is the first report on whole genome analysis of human G3P[3] RVA from China.
Child, Preschool ; China ; Genome, Viral ; Genomics ; Humans ; Male ; Molecular Sequence Data ; Phylogeny ; Reassortant Viruses ; classification ; genetics ; isolation & purification ; Rotavirus ; classification ; genetics ; isolation & purification ; Rotavirus Infections ; virology ; Viral Proteins ; genetics

AIM: To explore the long-term efficacy of Q-factor guided laser epithelial keratomleusis ( LASEK ) for myopia and astigmatism with positive Q-factor.
METHODS: There were 158 eyes which were myopia and astigmatism with positive Q- factor taken in two groups randomly: 86 eyes accepted Q - factor guided LASEK as observation group and 72 eyes accepted routine LASEK as control group. The difference between the two groups about all data was similar. The uncorrected visual acuity ( UCVA ) and the best corrected visual acuity ( BCVA ) as well as diopter, ocular tension, corneal topography, Keratometry value K, aspherical factor Q, Higher-order aberrations ( HOA ) , corneal thickness by ultrasound and, contrast sensitivity ( CS ) , Haze were examined and compared before and after surgery. All the cased were followed up for 14d, 1, 3, 6, 12mo. And there were no statistical difference among the data before surgery.
RESULTS: After 12mo there were no statistical difference between the two groups about UCVA and BCVA. But the safety index of observation group was 1.10, that of control group was 1. 07. The validity index of observation group was 1. 06, that of control group was 0.99. The HOA of observation group was 0. 45±0. 17μm, and that of control group was 0. 72±0.25μm, there was statistically significant difference (t=-8. 193,P=0. 000). Q factor of observation group was 0. 41±0. 17, that of control group was 0. 77±0. 22, there was significant difference (t=11. 377,P = 0. 028). The contrast sensitivity of 3mo post surgery of patients in the observation group returned to the level of before surgery. But in the control group the contrast sensitivity of the patients did not returned until 6mo.
CONCLUSION:Q-factor guided LASEK for myopia and astigmatism with positive Q-factor is stable, safe and effective. The operation allow for reducing the high order aberrations, maintaining the most asphericity of cornea, saving more in corneal tissue, which cause faster recovery of contrast sensitivity, less haze and better visual quality.

Heterokaryon incompatibility is a widespread phenomenon among fungi,controlled by specific loci termed het (for heterokaryon incompatibility).This review focuses on recent developments in our understanding of the molecular mechanisms of nonself recognition and the relationship between the death progresses of heterokaryon incompatibility and associated proteins in fungi.The deep research of heterokaryon incompatibility mechanism will hopefully reveal underlying principles of the evolution of nonself recognition systems and will find some effective method for settling the instability of protoplast fusant of fungi.

Objective To evaluate the utility of fluorescent dye SYTO 13 for high -resolution melting ( HRM) detection in single nucleotide polymorphism ( SNP) genotyping and its clinical application . Methods This is a performance verification study .36 genotype defined samples were divided into three groups:SNP rs3125734 C>T (class Ⅰ SNP) ,rs255758 A>C (class ⅡSNP) and rs688C>T.These samples were used to evaluate SYTO 13′s SNP genotyping capability of class ⅠSNP, classⅡSNP, and two PCR products of different lengths (52 and 107 bp) covering the same SNP of rs688C>T.The commercial HRM dye of LCGreen Plus was used as the control .The genotyping capability is indicated by the Tm difference(ΔTm) between wild type and homozygous mutant genotypes .The Tm differences between wild genotype and homozygous mutant genotype were compared using the Independent Samples t test.Paired t test was used to evaluate genotyping capability of the two dyes .The clinical applicability is evaluated by synchronously performing PCR amplification and HRM analysis on thirty -five randomly selected DNA samples with known genotypes of the three SNPs .Results The SNPs of class Ⅰ and class Ⅱ can be genotyped directly and clearly with SYTO13 (ΔTmclas Ⅰ =0.36 ±0.05,tclas Ⅰ =14.827,Pclas Ⅰ =0.000;ΔTm clas Ⅱ =0.42 ±0.110,tclasⅡ =9.539,Pclas Ⅱ =0.000).The classⅠSNP genotyping results was better using SYTO13 (ΔTmSYTO13 =0.39 ±0.027), while the SNP genotyping for small amplicon did not discriminated clearly in this study .Long amplicons of class ⅠandⅡSNPs can be identified directly except for several samples which can be genotyped accurately after having performed reexamination .Conclusion SYTO13 can apply for HRM analysis of genotyping classⅠand ⅡSNPs with long amplicon and for clinical routine detection.