AIM: To observe the efficacy of intravitreal injection with ranibizumab combined with laser photocoagulation for macular edema ( ME ) secondary to macular branch retinal vein occlusion( MBRVO) .
METHODS:A retrospective analysis included 33 patients (33 eyes) with ME secondary to MBRVO were taken. All patients received intravitreal injection of 0. 5mg ranibizumab ( 0. 05ml ) at first visit. The continue PRN treatment and laser photocoagulation were based on the visual acuity changes and optical coherence tomography findings. The changes of best corrected visual acuity ( BCVA) , central macular thickness( CMT) , and amplitude density and latency of P1 wave in mfERG were observed before treatment and 6mo after treatment.
RESULTS: Before the treatment, logMAR was 0. 68±0.35, 6mo after treatment was 0. 34±0. 23, BCVA was improved obviously ( P < 0. 01 ), BCVA in 21 patients ( 63.64%) were improved in two rows among all the patients. CMT before treatment was(487. 30±63. 58) μm, after treatment was(238. 84±52. 66) μm(P<0. 01). The amplitude densities of P1 wave in ring 1, ring 2 and ring 3 after treatment were significantly increased(all P<0. 01), and the latencies were decrease ( all P < 0. 05 ). The conjunctival hemorrhage was observed in 2 eyes after treatment.
CONCLUSION: Intravitreal injection with ranibizumab combined with laser photocoagulation for ME secondary to MBRVO can reduce the CMT and improve visual function.

Background Retinal ischemia reperfusion injury (RIRI) is a common pathological process of many retinal vascular diseases with comprehensive pathogenesis mechanism.Researches showed that apoptosis of retinal cells and nerve fiber loss is the finally common pathway of RIRI,and Janus kinase signal transducer and activator of transcription (JAK-STAT) pathway is a newly discovered signal transcript channel in recent years,which is involved in varieties of pathological processes.However,whether JAK-STAT pathway is associated with RIRI is still unelucidated.Objective This study was to investigate the time course of activation of JAK-STAT signal pathway and its significance during RIRI.Methods Forty clear adult male Sprague-Dawley rats were randomized into RIRI 6-hour,12-hour,24-hour and 48-hour groups.RIRI models were induced in lateral eyes of the rats by perfusing normal saline solution into the anterior chamber to elevate intraocular pressure (IOP) to 110 mmHg for 60 minutes and then allowing reperfusion,and the fellow eyes of the rats served as normal control group.The rats were sacrificed and the eyeballs were enucleated at 6,12,24 and 48 hours after reperfusion.The expressions of JAK2 and STAT3 protein (absorbance) in the retinas were located and detected by immunohistochemistry,and the relative expression levels of JAK2 and STAT3 mRNA in the retinas were detected by real-time fluorescence quantitative PCR.The use and care of the rats followed the ARVO Statement.Results Immunohistochemistry showed that JAK2 and STAT3 were faintly expressed in inner nuclear layer and retinal ganglion cells (RGCs) in the normal control group and strongly expressed in various RIRI groups.Significant differences were found in the expression intensities of JAK2 and STAT3 protein among the five groups (F =88.735,96.625,both at P ＜ 0.01).Compared with the normal control group,the expression intensities of JAK2 and STAT3 were enhanced in RIRI groups,with the peak values in RIRI 12-hour group (JAK2:t=4.308,5.559,5.315,4.726;all at P＜0.01.STAT3:t=5.047,7.843,6.281,4.887;all at P＜0.01).The thickening of inner retinal layer,loosening of retinal tissue,vacuolus degeneration of cells and decrease of RGCs were seen in the RIRI eyes.The relative expressing levels of the JAK2 mRNA and STAT3 mRNA in the retinas were significantly different among the groups (F =111.239,129.539;both at P＜0.01),and the relative expressing levels of JAK2 mRNA and STAT3 mRNA in the retinas were significantly increased in RIRI 6-hour,12-hour,24-hour and 48-hour groups in comparison with the normal control group (JAK2 mRNA:t=3.504,5.102,4.679,4.213;all at P＜0.01.STAT3 mRNA:t =6.541,8.787,5.693,5.898;all at P＜0.01).Conclusions The retinal morphology appears to be abnormal and RGCs are evidently decreased in rat eyes with RIRI,and the expressions of JAK2 and STAT3 in the retinas are simultaneously up-regulated,indicating that JAK-STAT signal pathway is involved during the RIRI process.

Objective To discuss the effects and influence of preoperative and postoperative adjunctive intravitreal conbercept in 23G vitrectomy for proliferative diabetic retinopathy (PDR).Methods A retrospective research was performed on 42 PDR eyes from January 2015 to February 2016 in the Second Affiliated Hospital of Nanchang University,who received either intravitreal 0.05 mL conbercept injection 7 days before 23G vitrectomy (group A,n =22) or intravitreal 0.05 mL conbercept injection at the end of 23 G vitrectomy (group B,n =20).The operative time,postoperative vitreous hemorrhage (VH),intraoperative and postoperative other differences of clinical indicators and postoperative best-corrected visual acuity (BCVA) between the two groups were compared.Results The average operation time,intraoperative electric coagulation hemostasis rate,iatrogenic hiatal incidence and intraoperative hemorrhage rate of group A were lower than those of group B (all P ＜ 0.05).BCVA at 6 months after surgery did not differ significantly between two groups (P ＞ 0.05),but the difference was statistically significant between pre-operation and post-operation (P ＜ 0.05).The incidences of early (≤ 1 month) postoperative VH were 18.2％ (4 eyes) and 15.0％ (3 eyes) in group A and B,respectively (P ＞ 0.05).The incidences of later (＞ 1 month) postoperative VH were 27.3％ (6 eyes) and 0 in group A and B,respectively,the difference was statistically significant (P ＜0.05).The percentages of reoperation were 13.6％ (3eyes with postoperative VH) and 10.0％ (2 eyes with traction retinal detachment) respectively in group A and B.The average times of supplementary laser photocoagulation were (2.3 ± 1.0) times and (1.4 ±0.6) times in group A and B,respectively in follow-up period (P ＜ 0.05).Conclusion The adjunctive use of intraoperative intravitreal injection of conbercept can prevent effectively postoperative VH and decrease conveniently the time of supplementary laser photocoagulation in 23G vitrectomy for PDR,as well as the preoperative adjunctive use can decrease the operation time,intraoperative complications and incidences of early postoperative VH.

To identify whether a highly repeated GT sequence from DCA1 promoter from Dunaliella salina,which have been proved to be a salt-inducible promoter in our previous study,would be a salt-inducible regulation element,different primers were designed to amplify 6 different-length fragments of DCA1 promoter from D.salina by PCR.After these fragments were respectively inserted into the HindⅢ-BamH I sites of the vector pU?GUS,serial expression vectors containing the gus gene were generated.D.salina cells transformed with these recombinant plasmids by electroporation were grown in liquid media containing different concentrations of sodium chloride respectively.GUS enzyme activity was measured histochemically and fluorometrically.The results revealed that 3 fragments containing GT repeated sequence drove the external gus gene expression and the expression pattern of the gus gene was regulated by the concentrations of sodium chloride.Additionally,the 2 fragments without tandem GT sequence drove the gus gene expression,but the expression pattern of the gus gene wasn't regulated by the concentration of sodium chloride;Also,the upstream fragment of the tandem GT sequence wasn't able to drive the gus gene expression.In conclusion,the highly repeated GT sequence from the DCA1 promoter plays an important role in the salt-inducible regulation of DCA1 promoter from D.salina and might be a novel salt-inducible element.

To discuss the influence of dexamethasion on lL-1β and TNF - α expression in suture - induced rabbit corneal neovascularization ( CNV ) and analyze the potential mechanism.
●METHODS: For 43 healthy rabbits, 40 were randomly selected for establishing CNV model in corneal stroma. The right eyes (group A) were received no medicine and the left eyes ( group B) were injected dexamethasone after successfully establishing the model. The no modeling 3 rabbits were normal control group. The morphologic change of corneal was observed with slit lamp microscope and the areas of CNV was calculated every day, then 8 rabbits were randomly chosen for sacrificing at 1, 4, 7, 14, 21d respectively. The pathological characteristics of CNV were observed after HE staining, and lL - 1β and TNF - α expression was detected by immunohistochemistry.
●RESULTS: CNV was grown at the 4d after suture, and the 7-14d was vigorous growth period. inflammatory cell infiltration appeared after HE staining, and CNV was located at the superficial stroma of cornea. lmmunohistochemistry results showed that lL - 1β and TNF - α expression was gradually increased with prolonged suture time. Compared with corneal stitch group, the rabbits cured by dexamethasone were found with less inflammatory cells infiltrating and neovescularization, moreover, the expression of lL - 1βand TNF-α decreased. There were statistical significance between the two groups (P<0. 05).
● CONCLUSlON: Dexamethasone can inhibit the CNV growth by controlling the inflammation of corneal and restraining lL-1β and TNF-α expression.

Objective: To research the best processing method for ginger pinellia by orthogonal tests.Methods: The orthogonal tests included the soaking time, boiling water and cooking time as the influencing factors, an HPLC method was used for the determination of 4 nucleosides (uridine, guanosine, adenosine, inosine), and the alum limit and extract content were also studied.The results were evaluated by multi index comprehensive weighted score to optimize the processing technology of ginger pinellia.Results: The best processing technology of ginger pinellia was as follows: soaked for 60 hours, the proportion of boiling water and pinellia tuber was 15:1, and boiled for about 5 h.Conclusion: The optimum processing technology of ginger pinellia is reasonable, reliable and reproducible, which can be used as the reference for the processing standardization of Chinese crude drugs.

Objective To prepare the standard molecular weight fragment mixtures. Methods Primers were designed to prepare clones which contained different sizes of standard molecular weight fragments. The template used for amplification of insert fragments was the pMD18-T vector. Bacteria culture and plasmid extraction were used to obtain abundant target fragment. Unlabeled DNA fragments were prepared by double digestion of the recombinant plasmids, and the fluorescent adaptor was prepared by annealing with two partial reverse complimentary DNA fragments. The unlabeled fragments and fluorescent adaptor were connected by DNA ligation reaction assisted with T4 DNA ligase. In this way, different sizes of standard molecular weight fragments were prepared. Standard molecular weight fragment mixture was finally prepared by mixing all the fragments together before purification. Results Ten standard molecular weight fragments of different sizes were prepared. The sizes of each fragment are 80bp, 124bp, 194bp, 224bp, 254bp, 304bp, 349bp, 399bp, 424bp and 454bp. The internal standard could accurately determine the size of PCR products amplified with the DNATyper15 kit. Conclusion Using this method, the standard molecular weight fragment mixture which meet the requirements of research and laboratory use was prepared, perfectly providing a new method for preparation of the DNA molecular weight standards. The peaks and the size of the prepared DNA internal lane standard are correct, which can be used to calculate the DNA fragments size in capillary electrophoresis.

Objective To explore the clinical efficacy of laparoscopic inguinal hernia repair (LIHR) in elderly patients.Methods The retrospective cohort study was adopted.The clinical data of 3 203 patients with inguinal hernias (3 847 sides) who were adnitted to the Ruijin Hospital of Shanghai Jiaotong University School of Medicine between January 2001 and December 2013 were collected.Of 3 203 patients,979 (1 107 sides) with age ＜ 60 years and 2 224 (2 740 sides) with age ≥ 60 years were respectively allocated into the under 60 years group and 60 years or older group.The surgical procedures including transabdominal preperitoneal (TAPP) approach,total extraperitoneal (TEP) approach and intraperitoneal onlay mesh (IPOM) approach were selected and performed by doctors in the same team.There were light-weight and heavy-weight patches.Observation indicators included (1) overall operation situations,(2) surgical comparison between the 2 groups,(3)comparison of postoperative indicators between the 2 groups,(4) follow-up.Follow-up using telephone interview and outpatient examination was performed to detect the recovery time of non-restricted activity,recurrence of hernia and complications.Measurement data with normal distribution were represented as ~ ± s and comparison between groups was done by the t test.Comparisons of count data were analyzed using the chi-square test or Fisher exact probability.Ranked data were compared by the nonparametric rank sum test.Results (1) Overall operation situations:3 203 patients with inguinal hernias (3 847 sides) underwent LIHR,including 1 475 (1 677 sides) using TAPP approach,1 718 (2 154 sides) using TEP approach and 10 (16 sides) using IPOM approach (6 using TAPP and IOPM approaches in each side).The light-weight patch was used in 2 206 sides and heavy-weight patch was used in 1 641 sides.Operation time was (31 ± 12) minutes in all 3 203 patients,(27 ±9)minutes in 2 559 patients with unilateral hernia and (44 ± 12)minutes in 644 patients with bilateral hernia,respectively.Duration of postoperative hospital stay was (1.5 ± 1.2) days.(2) Surgical comparison between the 2 groups:TAPP approach,TEP approach,IPOM approach,light-weight patch and heavy-weight patch were performed to 567,538,2,751,356 sides in the under 60 years group and 1 110,1 616,14,1 455,1 285 sides in the 60 years or older group,respectively,with statistically significant differences in above indicators between the 2 groups (X2 =37.976,70.022,P ＜ 0.05).Operation time in unilateral hernia and bilateral hernia and total operation time were (27 ± 9)minutes,(42 ± 10)minutes,(29 ± 10)minutes in the under 60 years group and (27 ± 10)minutes,(44 ± 12)minutes,(3 1 ± 13)minutes in the 60 years or older group,respectively,with no statistically significant difference between the 2 groups (t =-0.106,-1.768,-4.445,P ＞ 0.05).(3) Comparison of postoperative indicators between the 2 groups:the pain score at postoperative day 1 and duration of postoperative hospital stay were 2.4 ± 1.1,(1.5 ± 1.1) days in the under 60 years group and 2.3 ± 1.0,(1.5 ± 1.3) days in the 60 years or older group,respectively,with no statistically significant difference between the 2 groups (t =1.419,-0.126,P ＞0.05).(4) Follow-up:all the patients were followed up for 23-60 months,with a median time of 43 months.Cases with non-restricted activity recovery at postoperative week 2 and 4 were 973,978 in the under 60 years group and 2 208,2 222 in the 60 years or older group,respectively,showing no statistically significant difference between the 2 groups (X2=0.113,P ＞0.05).The recurrence of hernia,severe complications,serum tumescence,paresthesia and enteroparalysis were detected in 1,0,49,5,1 sides in the under 60 years group and 11,3,132,16,2 sides in the 60 years or older group,respectively,with no statistically significant difference between the 2 groups (x2=1.556,0.269,0.254,P ＞ 0.05).The urinary retention in the under 60 years group and 60 years or older group was respectively detected in 6 and 44 sides,showing a statistically significant difference between 2 groups (x2=6.956,P ＜ 0.05).Conclusion LIHR is safe and effective in elderly patients,and it can achieve good clinical efficacy under selecting reasonable operation procedures and patches.

Aim: Clone and characterize of the 5′- flanking region of the nitrate reductase (NR) gene derived from Dunaliella salina(D. salina). Methods : The genomic DNA from D. salina was respectively digested with BamHI, EcoRI, HindIII, Pst I, Sal I and Xba I. A genomic walking cassette was ligated to the ends of the digested DNA fragments, and then genomic walking libraries comprising BL, EL, HL, PL, SL and XL were constsucted. The 5′- flanking region of the NR gene from genomic walking libraries of D. salina was amplified by LA-PCR. The DNA sequences were analyzed with the software - Promoter Predictions. Isolated 5′-flanking regions fused to the GUS gene were tested for transient expression in the alga. Results: A single specific PCR product of about 1200bp in length from the HL library was generated. Also, several conserved motifs, such as CAAT-box, GAGA-box were found, which are related to regulation of transcription, and the putative binding sites of transcriptional factors such as EBP, EFII, NF-E1 and LV. BLAST showed that the DNA sequences shared high homology with 5′-upstream region of the NR gene from Dunaliella viridis. The isolated 5′-flanking regions were able to strongly drive GUS reporter gene expression, suggesting that it contains the promoter elements necessary for the transcription of the NR gene. The expression pattern of the GUS gene and the NR gene were similar, both ware induced by nitrate and repressed by ammonium. Conclusion: The cloned 5′- flanking sequences of NR gene derived from D. salina might be a specific promoter with the ability to“switch on or off” an expression of the heterologous gene in transgenic D. salina.

The purpose was to compare the difference between transgene expressions driven by homologous duplicated carbonic anhydrase (DCA) promoter and foreign CaMV35S promoter in the unicellular green alga, Dunaliella salina(D.salina).The CaMV35S promoter-bar construct and DCA promoter-bar construct into D.salina by a Backon 2000 electroporation system were introduced. After the repeated selections with the phosphinothricin (PPT) of 3mg/L, 3 PPT-resistant phenotype transformants were isolated from the CaMV-bar and DCA-bar pools of transformants of D. salina, respectively. The results of PCR and sequencing showed that bar genes were stably integrated into the genome of D.salina, and Southern bolts showed the number of transgene copy had no significant difference between both promoters. Semi-quantitive RT-PCR indicated that the mRNA levels of bar gene were higher in DCA-bar transformants than the CaMV-bar transformants, and could be increased under the induction of high salt in DCA-bar transformants but not in the CaMV-bar transformants. Analysis of growth rate of transformants showed DCA-bar transformants achieved the log stage faster than the CaMV-bar transformants. It is concluded that the homologous promoters have more advantages than the foreign promoters in the transgenic D.salina.