1.Recent advance about cytokines of proliferative vitreoretinopathy
Chinese Journal of Experimental Ophthalmology 2016;34(8):765-768
Proliferative vitreoretinopathy (PVR) is a leading cause of blindness and caused by abnormal repairs for the damaged vitreous and retina.The formation of PVR involves many cytokines,including platelet derived growth factor (PDGF),vascular endothelial growth factor,hepatocyte growth factor,transforming growth factor-β,epidermal growth factor,tumor necrosis factor-α,connective tissue growth factor,etc.The Jagged-1/Notch signaling pathway and RhoA/ROCK signaling pathway are involved in the epithelial-mesenchymal transition of retinal pigment epithelium (RPE) cells.The migration and proliferation of RPE cells are associated with the PKC/ERK and Akt/mTORCl signaling pathway.Non-PDGF cytokines indirectly active PDGF receptor α,which seems to be the key pathway in the development of PVR.The methods of cocktail neutralizing reagents targeted to multiple cytokines and the antioxidant N-acetylcysteine are effective in preventing PVR.Here,we reviewed the mechanisms and signaling pathways of multiple relevant cytokines involved in development and treatment of PVR.
3.Mutation analysis of Pax6 in Chinese patients with congenital aniridia
Peng, HAO ; Ming, YING ; Ruifang, HAN ; Liming, WANG ; Ningdong, LI
Chinese Journal of Experimental Ophthalmology 2016;34(10):900-904
Background Congenital aniridia is a rare congenital autosomal dominant disease,which is shown as aniridia of double eyes,and the paired box gene 6 (Pax6) gene mutation is now known to be associated with congenital aniridia.Objective This study was to screen the Pax6 gene mutation in patients with congenital aniridia.Methods Eleven patients with congenital aniridia were enrolled in Tianjin Eye Hospital from August 2012 to October 2015,including 6 patients from 3 congenital aniridia family and 5 sporadic patients.All patients received routine ophthalmic examination.Peripheral venous blood of 3 ml was collected from the patients for DNA extraction according to the standard process of DNA isolation instructions,and all the exons of Pax6 gene,Elp4 gene,exon 5 ' and 3',intron splice sequence and SIMO sequence were amplified by PCR.Pax6 genes of the patients were sequenced using Sanger direct sequencing and multiplex ligation dependent probe amplification (MLPA) and compared with those of 500 ocular trauma patients.This study complied with Helsinki declaration,and written informed consent was obtained from each patient prior to any medical examination.Results Iris absence was found in all the patients,and the visions acuity was hand motion to 0.2.Lens dislocation was seen in 1 patient.Direct sequencing results found that three patients in AN-O1 family were c.688g>t (p.E230X) mutation of Pax6 gene,and 3 of 5 sporadic patients carried c.468g>a (p.W156X),c.613c>t (p.Q205X) and c.141 +2t>c mutant of Pax6 gene,and the c.688g>t (pE230X) mutation was a novel-discovered mutation.No any mutation in Pax6,Elp4 gene and SIMO fragment was detected in 1 patient from AN-02 family,2 patients from AN-03 family and 2 sporadic patients by both direct sequencing and MLPA validation.No above-mentioned mutation was found in 500 normal individuals.Conclusions The mutation of Pax6 gene is a pathogenic mutation in congenital aniridia patients,and c.688g>t (p.E230X) is a novel Pax6 mutant,which expanded the mutation spectrum of Pax6 gene.
4.Screening of TYR gene mutations and clinical classification in oculocutaneous albinism patients
Liming, WANG ; Ruifang, HAN ; Ming, YING ; Peng, HAO ; Ningdong, LI
Chinese Journal of Experimental Ophthalmology 2016;34(10):905-909
Background Oculocutaneous albinism (OCA) is a hereditary disease of pigment absence in eyes,skin and hair due to the lack of congenital melanocyte.OCA is classified into 7 types based on different genetic mutations,and the mutation of tyrosinase (TYR) gene causes OCA type 1 (OCA1).OCA has obvious genetic heterogeneity and phenotypic heterogeneity.The molecular diagnosis of the mutant gene is helpful for the classification and molecular pathogenesis study of OCA.Objective This study was to screen the TYR mutation in OCA patients,and to analyze the association between the gene mutation type and clinical phenotype.Methods Ten patients with OCA were enrolled in Tianjin Ophthalmological Hospital from January 2011 to December 2014.The clinical and ocular manifestations of the patients were examined.Peripheral venous blood 3 ml was collected in the patients and their lineal relatives for the extraction of genomic DNA.Extracted DNA was amplified by PCR and the TYR gene sequence was analyzed,including all 5 exon coding sequence and exon 5 ' and 3' end and the non-coding region sequence of intron splicing in TYR gene.This study complied with Helsinki Declaration and the protocol was approved by Ethic Committee of Tianjin Eye Hospital.Informed consent was obtained from each subject.Results All the patients showed white or reddish hair and snow-white skin,and different degrees of pigment lack was seen in iris.The best corrected visual acuity of the patients was 0.05-0.2,and 3 patients complicated with nystagmus.Fundus findings showed a sunset-like change and dysplasia of macula.The TYR gene sequencing revealed that patient 1 was OCA1A subtype,with the compound heterozygous mutant of c.832C>T (p.R278X) and c.1217C>T (p.P406L),and his/her parents occurred the heterozygous mutation of exons P406L and R278X.The phenotype of the patient 1 was white hair and white iris.The patient 3 was OCA1B subtype,with the compound heterozygous mutations of c.1265G>A (p.R422Q) and c.1217C>T (p.P406L),showing an appearance of reddish brown hair and sallow iris.TYR gene mutant was not detected in other 8 patients.Conclusions The mutation of TYR gene is the main cause of OCA1 type.The phenotype of OCA1A subtype is no pigment in eyes and hair,and one of OCA1B subtype was obviously lessening of pigment.The difference of mutant genes of OCA is the cause of genetic and phenotypic heterogeneity.
5.Expression of VEGF -C and p63 in early esophageal carcinoma and intraepithelial neoplasia
Songyan HAN ; Li LI ; Ruifang SUN ; Suhong LI ; Ziyan HAN ; Ming ZHAO ; Quanhong WANG
Cancer Research and Clinic 2009;21(11):762-764,771
Objective To explore expression and clinical value of VEGF-C and p63 in early esophageal carcinoma and intraepithelial neoplasia. Methods 146 cases were randomized into normal esophageal mucosa, low level intraepithelial tumor, high level intraepithelial tumor and early esophageal carcinoma. The expression of VEGF-C and p63 were detected by using the immunohistochemistry dyeing.Results The expression of VEGF-C immunohistochemistry dyeing had statistical differences among different levels(X~2= 47.455, P <0.001). Normal esophageal mucosa v.s. high level intraepithelial tumor (X~2=36.721, P <0.001), Normal esophageal mucosa v.s. early esophageal carcinoma (X~2=26.483, P <0.001), low level intraepithelial tumor v.s. high level intraepithelial tumor(X~2= 10.025, P<0.0083), low level intraepithelial tumor v.s. early esophageal carcinoma(X~2=16.734, P<0.001). There was a significant correlation between pathological classification and the expression amount of VEGF-C (r = 0.462, P <0.001). The expression of p63 had statistical differences among different levels(X~2=28.962, P <0.05). There was a significant difference on normal esophageal mucosa comparing with low level, high level intraepithelial tumor or early esophageal carcinoma (X~2=12.735, P =0.005, X~2=20.421, P<0.001, X~2=20.854, P<0.001). There was a significant correlation between pathological classification and the expression of p63 (r= 0.272, P<0.05). Conclusion There is a significant correlation in the express of either VEGF-C or p63 comparing with either intraepithelial tumor or early esophageal carcinoma. It may be an early warning indicator.
6.Application of three-dimensional power Doppler in diagnosing and treating gestational trophoblastic disease
Wei WANG ; Zhen HAN ; Xueye TIAN ; Ting ZHANG ; Yanyan WANG ; Ruifang AN
Journal of Xi'an Jiaotong University(Medical Sciences) 2015;(6):819-823
Objective To clarify the reliability of three-dimensional power Doppler as a quantitative method in detecting gestational trophoblastic disease (GTD ) and assessing its therapeutic effect. Methods We prospectively collected a database of 52 patients with GTD diagnosed at the First Affiliated Hospital of Xi’an Jiaotong University from December 201 1 to October 2013 and 30 healthy women as controls.Sonography was performed using a Voluson E8. Resistance index (RI ), vascularization index (VI ), flow index (FI ) and vascularization-flow index (VFI)of the region of interest were collected for further analysis.Results Of the 52 GTD datasets,variation from the mean value was RI 0.47±0.1 7;VI (81.46 ±20.54)%;FI 67.28 ±20.21;and VFI 58.12±25.53.Three-dimensional power Doppler examination indicated that there were significant differences in RI,VI,FI and VFI values between healthy individuals and patients in each subgroup (P <0.01).Further,after combining invasive hydatidiform mole and choriocarcinoma groups,it also showed a significant difference between hydatidiform mole group and the combined malignant group (P <0.01).And the abnormal sonographic and power Doppler findings in GTD were resolved when chemotherapy was given successfully.Conclusion RI,VI,FI and VFI values were more related to the information of tumor blood flow,and more intuitive to manifest the status of the disease.They could be new methods in diagnosing and assessing treatment of GTD for their real-time nature.
7.The investigation of high risk type human papillomavirus prevalence and incidence rates of cervical intraepithelial neoplasia in Shenzhen women with different screen
Ruifang WU ; Qingzhi ZHOU ; Zhihua LIU ; Ruizhen LI ; Chunyan HAN ; Jufang SHI ; Junhua LI ; Lijie ZHANG ; Ni LI ; Youlin QIAO
China Oncology 2006;0(09):-
Background and purpose:HPV infection is known as the primary cause of cervical cancer worldwide To investigate high risk type human papillomavirus (HPV) prevalence and incidence rates of cervical intraepithelial neoplasia (CIN) and their screen risk factors in women with different methods of screening.Methods:1137 residents, workers and service women aged 15-59 from Shenzhen city were investigated for cervical cancer in an epidemiology screening study.The high risk types of human papillomavirus of liquid-based cytology samples were tested by hybrid capture 2 (HC-Ⅱ) and liquid-based cytology test (LCT) was also performed at the same time. Women for HPV-positive with LCT ≥ atypical squamous cells of undetermined sign (ASCUS) or HPV-negative with LCT ≥ low grade squamous intraepithelial lesion (LSIL) were biopsied in colposcopy and then were examined by pathology. All data was managed by Foxbase. ?2 test and unconditional Logistic regression model were used for data analysis by SPSS 10.0.Results:1137 women were eligible in our research, the overall rates of HPV infection was 14.0%. HPV detection rates in residents, workers and service women were 14.1%,9.2%,18.9% respectively. HPV detection rates in workers group was significantly lower than that of service women and residents (P
8.Establishment of real-time fluorescent quantitative PCR for detection of torque teno virus types 7, 8 and 10
Zhiqiang XIA ; Juan SONG ; Dong XIA ; Qinqin SONG ; Wenjun WANG ; Ruifang WANG ; Bingtian SHI ; Mi LIU ; Geng HU ; Yanhai WANG ; Jun HAN
Chinese Journal of Microbiology and Immunology 2021;41(3):190-194
Objective:To establish a real-time fluorescent quantitative PCR for the detection of torque teno virus types 7 (TTV7), 8 (TTV8) and 10 (TTV10) and analyze its performance in clinical sample detection.Methods:Specific primers were designed based on the gene sequences of TTV7, TTV8 and TTV10 in GenBank. Recombinant plasmids of pMD19-T-TTV7, pMD19-T-TTV8 and pMD19-T-TTV10 were constructed and used as positive standard control to establish a real-time fluorescent quantitative PCR based on FAM-Eclipse probe method. The specificity and sensitivity of the established method were evaluated. Moreover, it was validated in terms of clinical sample detection.Results:The standard curve equations of the real-time fluorescent quantitative PCR for detecting TTV7, TTV8 and TTV10 were y=-0.340 2 x+ 114.780 0 ( R2=0.998 8), y=-0.351 1 x+ 114.940 0 ( R2=0.995 3) and y=-0.348 9 x+ 115.020 0 ( R2=0.991 7), respectively, and there was no cross-reaction with other viruses. The detection sensitivity of the established method for TTV7, TTV8 and TTV10 were 108 copies/μl, 84 copies/μl and 98 copies/μl, and the positive detection rates in clinical pediatric serum samples were 10.9%, 2.1% and 4.3%, respectively. Conclusions:The established real-time fluorescent quantitative PCR for detection of TTV7, TTV8 and TTV10 was featured by strong specificity and high sensitivity, which could be used for rapid TTV detection in clinical serum samples.
9.Modified strategy of anesthesia for thoracoscopic radical resection of lung cancer: serratus anterior plane block combined with general anesthesia
Liang LIU ; Jianxu ER ; Ruifang GAO ; Ying ZHANG ; Bingsha ZHAO ; Jiange HAN
Chinese Journal of Anesthesiology 2021;41(7):831-834
Objective:To evaluate the modified efficacy of serratus anterior plane block (SAPB) combined with general anesthesia for thoracoscopic radical resection of lung cancer.Methods:Eighty-two patients of both sexes, aged 40-64 yr, with body mass index of 18-24 kg/m 2, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, scheduled for elective thoracoscopic radical resection of lung cancer, were divided into 2 groups ( n=41 each) using a random number table method: general anesthesia group (group G) and SAPB combined with general anesthesia group (group SG). Ultrasound-guided SAPB was performed before induction of general anesthesia in group SG.General anesthesia was induced with midazolam, etomidate, sufentanil and cis atracurium, and anesthesia was maintained with sevoflurane and remifentanil.Sufentanil was used for patient-controlled intravenous anesthesia (PCIA) after the end of operation.When visual analog scale score≥4, sufentanil 2.5 μg was injected intravenously for rescue analgesia.The intraoperative consumption of sevoflurane and remifentanil, extubation time, requirement for rescue analgesia within 48 h after operation, consumption of sufentanil, requirement for nicardipine and esmolol and occurrence of adverse events were recorded. Results:Compared with group G, the intraoperative consumption of remifentanil and sevoflurane, postoperative consumption of sufentanil, postoperative requirement for rescue analgesia, postoperative requirement for nicardipine and esmolol, postoperative incidence of nausea and vomiting, skin pruritus and urinary retention were significantly decreased, the extubation time was shortened, and the time of the first postoperative requirement for rescue analgesia was prolonged in group SG ( P<0.05). Conclusion:Compared with general anesthesia alone, SAPB combined with general anesthesia can not only significantly reduce intraoperative general anesthetics and opioid consumption, but also improve postoperative stress management, which is helpful for early postoperative outcome when used for thoracoscopic radical resection of lung cancer.
10.Content variation of four flavonoid glycosides in Hippophae rhamnoides L .leav-es before and after fermentation assayed by RP-HPLC
Ruifang DONG ; Zheng WANG ; Ling CHEN ; Zhiheng SHU ; Ting HAN ; Luping QIN
Journal of Pharmaceutical Practice 2017;35(6):526-529
Objective To compare the changes of four flavonoid glycosides in the leaves of Hippophae rhamnoides L . before and after fermentation .Methods The water extract of Hippophae rhamnoides L .leaves and its fermented tea were con-centrated and desiccated .The dry extracts were dissolved in 70% ethanol .The chromatographic separation was performed with RP-HPLC method on an Extend-C18 column (4 .6 mm × 250 mm ,5 μm) .Acetonitrile-0 .1% formic acid was selected as mobile phase at the flow rate of 1 .0 ml/min .The detection wavelength was 356 nm and the column temperature was 30 ℃ .Results The rutin content was high in the leaves of Hippophae rhamnoides L .After fermentation ,isoquercitrin content was increased , while the contents of rutin and narcissoside were reduced and isorhamnetin-3-O-glucoside stayed unchanged .There was a good linear relationship between the concentration and peak areas of the four compounds (r>0 .9997) .The average recoveries were between 96%-103% .Conclusion This established method is rapid and reliable ,which can be used for the quality control of Hippophae rhamnoides L .leaves and its fermented tea .