Construction of yeast one-hybrid library and screening of transcription factors regulating FPS expression in Ganoderma lucidum / 中草药

Objective: To screen the upstream regulatory transcription factors of farnesyl diphosphate synthase (FPS) in the triterpenoid synthesis pathway in Ganoderma lucidum. Methods: In this study, the FPS promoter was cloned and connected to the pAbAi plasmid to construct bait vector pAbAi-FPS, which was transformed into Y1H yeast competent cells to construct bait yeast. The yeast one-hybrid cDNA library was constructed by using SMART technology, then the purified ds-cDNA and pGADT7-Rec were co-transformed into bait yeast strain to screen the upstream transcriptional regulatory factors of PFS. Results: The bait vector containing pAbAi-FPS was constructed and the bait strain was screened, the cDNA library was constructed and transformed to the bait strain. A total of 37 positive clones were screened and sequenced. The sequences of conserved domain were predicted and performed blast search against the whole-genome database to identify their function. As a result, a total of 18 upstream regulatory factors were screened out including three transcription factors, five ribosomal proteins, and 10 other transcription regulators. Conclusion: The results indicated that transcription factors GlSNF2, GlMHR, and GlZn2Cys6 were candidate genes for regulating the expression of FPS, and this study offered data for further study on the regulation mechanism of FPS expression.

Construction of yeast one-hybrid library and screening of transcription factors regulating LS expression in Ganoderma lucidum / 中国中药杂志

Lanosterol synthase( LS) is a key enzyme involving in the mevalonate pathway( MVA pathway) to produce lanosterol,which is a precursor of ganoderma triterpenoid. And the transcriptional regulation of LS gene directly affects the content of triterpenes in Ganoderma lucidum. In order to study the transcriptional regulation mechanism of LS gene,yeast one-hybrid technique was used to screen the transcription regulators which interact withthe promoter of LS. The bait vector was constructed by LS promoter,then the vector was transformed yeast cells to construct bait yeast strain. One-hybrid c DNA library was constructed via SMART technology. Then the c DNA and p GADT7-Rec vector were co-transformed into the bait yeast strain to screen the upstream regulatory factors of the promoter region of LS by homologous recombination. Total of 23 positive clones were screened. After sequencing,blast was performed against the whole-genome sequence of G. lucidum. As a result,8 regulatory factors were screened out including the transcription initiation TFIIB,the alpha/beta hydrolase super family,ALDH-SF superfamily,60 S ribosomal protein L21,ATP synthase β-subunit,microtubule associated protein Cript,prote asome subunit β-1,and transaldolase. Until now,the regulation effect of these 8 regulatory factors in G.lucidum has not been reported. This study provides candidate proteins for in-depth study on the expression regulation of LS.

Comparative study on genotype distribution of HPV infection in the tissues of cervical squamous cell carcinoma and cervical adenocarcinoma / 医学研究生学报

Objective There are few large sample data reports of comparative study on genotype distribution of human papillomavirus (HPV) infection in the tissues of cervical squamous cell carcinoma and cervical adenocarcinoma in China. This study aimed to investigate the clinical value of genotype distribution of HPV infection in the tissues of cervical squamous cell carcinoma and cervical adenocarcinoma in regional (mainly in Jiangsu Province) patients.Methods We collected 1044 paraffin tissue specimens of female cervical squamous cell carcinoma (826 cases) and cervical adenocarcinoma (218 cases) in 29 hospitals from November 1978 to December 2017. HPV DNA was extracted from the tissues and through the combination of gene-chip and polymerase chain reaction technology, 23 genotypes of HPV were detected in all the tissues of cervical squamous cell carcinoma and cervical adenocarcinoma, and comparative study was conducted on the genotype distribution of HPV infection.Results Among 1044 cases of cervical squamous cell carcinoma and cervical adenocarcinoma, 901 were found with HPV and the detection rate was 86.30%. The detection rate of cervical squamous cell carcinoma was 91.53% (756/826), among which 16，18，58，33，52，31 types were the most common and the detection rate of 16 type was significantly higher than that of 18 types (56.84% vs 9.93%, P0.05).Conclusion The HPV detection rates are different in the regional female cervical squamous cell carcinoma and cervical adenocarcinoma tissues. 16,18,31,33,52 and 58 types are the most common genotypes in cervical squamous cell carcinoma and cervical adenocarcinoma. The detection rate of 16 type is overly higher than that of 18 type in cervical squamous cell carcinoma, while the detection rates of 16 type and 18 type in cervical adenocarcinoma are very close.

Clinical observation of 15 cases of corneal foreign body injuries caused by chestnut thorns / 国际眼科杂志(Guoji Yanke Zazhi)

Abstract?AIM:To investigate the effective treatment methods of corneal injury caused by chestnut thorns and the factors affecting the disease progression.?METHODS: From Jul.2014 to Oct.2015, the clinical data of 15 patients(15 eyes) with corneal injury caused by chestnut thorns in Ophthalmology Inpatient Department of Wuhan Tongji Hospital was retrospective analyzed. The patients without fungal keratitis were treated with the surgery of removing chestnut thorn from cornea and antifungal drugs. For the patients complicated with fungal keratitis, besides surgery of removing chestnut thorn and antifungal drugs, anterior chamber irrigation and corneal stroma injection with fluconazole solution were given to treat the disease.If necessary, amniotic membrane transplantation or keratoplasty was also given to the patients complicated with fungal keratitis. After that, the effectiveness of those methods and the factors affecting progression were analyzed.?RESULTS:For 11 patients without fungal keratitis, the average time between corneal injury and receiving treatment at Tongji Hospital was 1-7 (2.42±2.15) d and for 4 patients complicated with fungal keratitis, the average time was 3-30 (18.25±4.35)d.Among 15 cases, statistics suggested that the average number of chestnut thorn in patients complicated with fungal keratitis was 4.5, and all the chestnut thorn penetrated the cornea into the anterior chamber.The average number of chestnut thorn in patients without fungal keratitis was 3.5, and the proportion of chestnut thorn penetrated the cornea into the anterior chamber was 28.5%.After treatment, all patients had no new fungal keratitis or other complications.Those results indicated that the different treatments for the patients with or without fungal keratitis were all effective.?CONCLUSION:The factors affecting the progression of cornea foreign body injury caused by chestnut thorn are the number of chestnut thorn, whether chestnut thorn penetrate the cornea into the anterior chamber, time since injury, active anti -fungal therapy. If patients complicated with fungal keratitis could be treated with antifungal agents and anterior chamber irrigation or corneal stroma injection using fluconazole solution without delay, the progress of fungal keratitis could be effectively controlled, and favorable conditions for further therapy such as amniotic membrane transplantation or keratoplasty could be provided.

Comparison of anterior segment parameters between fellow eyes of unilateral acute angle closure glaucoma and primary angle closure suspects / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To explore the differences of anterior segment parameters in the patients with fellow eyes of unilateral acute angle-closure glaucoma ( AACG ) , primary angle-closure suspects ( PACS) and normal group. METHODS: Twenty-six eyes of 26 patients with fellow eyes of AACG, 28 eyes of 28 age- and gender-matched PACS and 34 normal eyes were imaged using optical coherence tomography ( OCT) and pentacam scheimpflug system ( Pentacam ) . Anatomical parameters including central corneal thickness ( CCT ) , corneal volume ( CV ) , pupillary diameter ( PD ) , central anterior chamber depth ( CACD ) , peripheral anterior chamber depth ( PACD ) , anterior chamber volume ( ACV ) and anterior chamber angle ( ACA) were obtained from Pentacam. Iris thickness (IT750,IT2000), cross-sectional area (IS), volume (IV) and angle opening distance 500 (AOD500) were estimated using OCT combined with a computer image processing. Statistic analysis was performed with SPSS. RESULTS: There were no significant differences in corneal parameters (CCT, CV), PD and iris values (IT750, IT2000, IS, IV) among the three groups (P> 0. 05). Compared with the fellow eyes of AACG and PACS, normal eyes had larger ACV, wider AOD500 and ACA, deeper CACD and PACD ( P 0. 05). Using the fellow eyes of AACG as the standard to predict high risk of angle closure, areas under the receiver operating characteristic curve of the above parameters were all less than 0. 7. CONCLUSION:All the anterior segment parameters are no different significantly between the fellow eyes of AACG and PACS. They may be notaccurate criteria for determining high risk group of PACS.

Diagnostic performance of macular ganglion cell - inner plexiform layer thickness measured by Cirrus HD-OCT in POAG patients / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To evaluate the diagnostic accuracy of macular ganglion cell - inner plexiform layer ( GCIPL ) measurements using high- definition optical coherence tomography ( Cirrus HD - OCT ) ganglion cell analysis algorithm for detecting early and moderate to severe glaucoma. METHODS:Twenty normal control persons, 26 patients with early glaucoma and 29 patients with moderate to severe glaucoma were enrolled in this study. Macular GCIPL, optic nerve head ( ONH ) parameters and peripapillary retinal nerve fiber layer ( RNFL ) thickness were measured in each subject. Then all measured results of each parameter were calculated using SPSS17. 0. Areas under the receiver operating characteristic curves ( AUC) of each parameter were calculated to compare the diagnostic accuracy for detecting early and moderate to severe glaucoma. RESULTS: For detecting early glaucoma, AUC of average RNFL and seven clock value of RNFL were the biggest ( 0. 871 and 0. 896 respectively ), the AUC of parameters in GCIPL were also significant, among them, the average GCIPL showed bigger AUC(0. 847) than the minimum GCIPL (0. 812). For diagnosing moderate to severe glaucoma, the AUC of rim area was 0. 992, which was bigger than that of average RNFL ( 0. 991 ). The minimum GCIPL showed bigger AUC ( 0. 983 ) than the average GCIPL (0. 967). For early glaucoma diagnosis, the sensitivity of average RNFL was the highest (76. 9%), while the average GCIPL has the highest specificity (93. 5%). CONCLUSION: AS a new diagnostic parameter for detecting glaucoma, GCIPL shows similar diagnostic potential compared with RNFL. For early glaucoma diagnosis, average RNFL is the most important parameter, while screening early glaucoma, average GCIPL should be paid more attention.

Analysis of cell morphology and immunophenotypic characteristics in 47 cases of multiple myeloma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>This study was to investigate the cell morphology and cell immune phenotypic characteristics in patients with multiple myeloma (MM).</p><p><b>METHODS</b>The flow cytometry with multiparametric direct immunofluorescence technique, and CD45/SSC and CD38(+)(+)/CD138(+) gating were used to measure cell markers CD138, CD38, CD56, CD117, CD3, CD13, CD33, CD19, CD7, CD20, CD22, CD34, CD28 in 47 MM patients. At the same time the morphology examination of bone marrow cells was performed.</p><p><b>RESULTS</b>The suspicious myeloma cell ratio in MM patients was 9.42%-74.25% detected by flow cytometry, moreover, the myeloma cell ratio detected by morphology examination was 11.0%-80.6%, there was a good correlation between the two detection methods (r(2) = 0.54, P < 0.001). The ratio of antigen positive expression was as follows: 74.46% for CD138, 100% for CD38, 57.44% for CD56, 40.42% for CD117, 6.38% for CD13, 19.15% for CD33, 8.51% for CD20, 27.66% for CD28, 2.12% for CD22, 4.25% for CD34, 0% for CD3, 0% for CD19, 0% for CD7.</p><p><b>CONCLUSIONS</b>CD45/SSC and CD38(+)/CD138(+) gating technique can accurately gate multiple myeloma cell sets which need analysis, the majority of myeloma cells expreses CD138, CD38, CD56 antigens. The immunophenotypic analysis combined with the cell morphology examination more contribute to the diagnosis and differential diagnosis of multiple myeloma.</p>

Identification of Placenta hominis and its adulterants using COI barcode / 中国中药杂志

In order to provide a new method for the identification of Placenta hominis, the COI barcode has been employed to identify the P. hominis medicinal materials and its adulterants. Genomic DNA was extracted from the experimental samples. The COI sequences were amplified and sequenced bi-directionally. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner. NJ tree was constructed by MEGA6.0 software. COI sequences can be successfully obtained from all experimental samples. The intra-specific variation and inter-specific divergence were calculated. The average intra-specific K2P distance of P. hominis was 0.001 and the maximum intra-specific distance was 0.008. The cluster dendrogram constructed can be seen that the same genus is together, and distinguished from its adulterants. It is concluded that P. hominis and its adulterants can be correctly identified by DNA barcoding method.

Expression of CIP2A in non-small cell lung cancer and its clinical significance / 肿瘤

Objective: To investigate the expression of cancerous inhibitor of protein phosphatase 2A (CIP2A) in non-small cell lung cancer (NSCLC) tissues and its clinical significance. Methods: The expressions of CIP2A protein in NSCLC tissues and their corresponding para-cancerous tissues from 97 cases of NSCLC were detected by immunohistochemistry. The relationship between the expression of CIP2A protein and the clinicopathological features in patients with NSCLC was analyzed. Results: The positive expression rate of CIP2A protein in NSCLC tissues [76.3% (74/97)] was significantly higher than that in the corresponding para-cancerous tissues [5.2% (5/97)] (P 0.05). The overall survival time of patients with positive expression of CIP2A protein was shorter than that of patients with negative expression of CIP2A protein (P =0.001). The expression of CIP2A protein, TNM stage and the differentiation of tumor were independent prognostic factors of NSCLC (P <0.05). Conclusion: The expression of CIP2A protein is significantly increased in NSCLC tissues, and the positive expression of CIP2A protein is associated with the prognosis of patients with NSCLC. Copyright© 2011 by TUMOR.

Are leukemic patient bone marrow mesenchymal stem cells malignant? / 中国实验血液学杂志

The study was aimed to explore whether there are leukemic characteristics in the bone marrow mesenchymal stem cells (BMMSC) from leukemic patients as compared with normal controls. The mesenchymal stem cells from bone marrow of normal volunteers and patients with APL and CML were isolated, then cultured and proliferated in vitro. The morphology, growth curve and cell surface markers of two different sources mesenchymal stem cells were investigated for detecting whether the bone marrow mesenchymal stem cells derived from leukemia patients have the specific abnormal fusion gene of leukemia cells through fluorescent in situ hybridization. The results indicated that there was no significant difference between the mesenchymal stem cells derived from different subjects, the bone marrow mesenchymal stem cells derived from leukemia patients did not have the clonal malignant fusion gene as seen in the leukemia cells. Taken altogether, mesenchymal stem cells derived from leukemia patients had no biological differences as compared with those from normal volunteers, and no malignant clonal abnormality was found. It is concluded that mesenchymal stem cells derived from leukemia patients as an alternative vehicle may be used for assistant of autologous hematopoietic stem cell transplantation or cell therapy and gene therapy.

Comparison of the botanic morphology and blooming characteristics of four cultivars of rose / 中国中药杂志

<p><b>OBJECTIVE</b>To establish identifying method for further development and utilization by studying botanic morphology and blooming characteristics of four varieties of roses in Jiangsu province.</p><p><b>METHOD</b>Flower-bud and flower-form were observed by dissection and plant modality and blooming process were investigated.</p><p><b>RESULT AND CONCLUSION</b>The flower form and plant modality was obviously different among the 4 varieties of roses. The process of differentiation of flower-bud could be divided into five stages: the transformation of nutritive growth cone, the occurrence and development of sepal, formation of petal primordium, formation of pistil and stamen. The blooming process was made up of flower-bud period, display-petal period, initiating blooming period, blooming period, withering period and corresponding biological marks.</p>