Objective:To observe the 5-year outcomes and complications after cataract extraction combined with intraocular lens implantation in children with bilateral congenital cataract.Methods:An observational case series study was performed.A total of 72 children (144 eyes), aged 2 to 7 years, who received intraocular lens (IOL) implantation combined with posterior continuous curvilinear capsulorhexies and anterior vitrectomy after cataract extraction for bilateral congenital cataract in The First Affiliated Hospital of Zhengzhou University from January 2007 to January 2012 were enrolled.All the patients underwent refractive correction and amblyopia training two weeks after the second operative eye underwent surgery.The patients were divided into 2-3 years old group (64 eyes), 4-5 years old group (48 eyes) and 6-7 years old group (32 eyes) according to their age at surgery.The preoperative and postoperative best corrected visual acuity (BCVA), refraction, axial length, corneal curvature, binocular visual function and postoperative complications were observed during the 5-year follow-up period.The influence of age at surgery on postoperative BCVA in one eye at the final follow-up visit was analyzed.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of The Affiliated First Hospital of Zhengzhou University (No.2020-KY-219), and written informed consent was obtained from guardians of the subjects before treatment.Results:The mean myopic shift was -2.10(-2.90, -1.90), -1.73(-2.50, -2.10) and-0.52(-2.00, -0.28)D in the 2-3 years old group, 4-5 years old group and 6-7 years old group, respectively, and the axial elongation in the three groups was (1.41±0.32), (0.96±0.51), and (0.52±0.26)mm, respectively, and the differences among the three groups were statistically significant ( H=19.85, P<0.01; F=13.24, P<0.05).There was no statistically significant difference in preoperative and postoperative corneal curvature among the three groups (all at P>0.05).The BCVA was significantly improved in all the eyes after operation, and a certain binocular visual function was obtained, and the visual function in the 2-3 years old group was better than that of 4-5 years old group and 6-7 years old group (all at P<0.05).The BCVA at the end of following-up was 0.3 or better in 116 eyes (80.56%), and was better than 0.8 in 28 eyes (19.44%).The regression analysis results showed that children younger at surgery had better visual outcome (postoperative BCVA=0.959-0.104 operative age of patient; R2=0.539, P<0.01). Conclusions:The recovery of visual function in children with binocular congenital cataract is related to the age at operation.The younger the age at operation is, the better the recovery of visual function will be.Timely, systematic and standardized refractive correction, amblyopia treatment and binocular visual function training after operation are helpful to the development and reconstruction of visual function.

Background Research comfirmed that second mitochondrial activator of caspase (Smac) is a promoting tumor cell apoptosis protein.Our previous study showed that the expression level of Smac in LECs is obviously higher in cataract than that in normal eyes.We assumed that silencing Smac gene in LECs can inhibit the apoptosis of LECs.The way to transfect Smac siRNA into LECs is a key step.Objective This study was to construct siRNA lentiviral vector of Smac and identify its silencing efficiency in human lens epithelial B3 cell line (HLE-B3) and establish low-expressed Smac HLE-B3 line.Methods Based on the genebank and our previous study,siRNA sequence of Smac was designed and composed.The synthetic double-stranded DNA was linked to the lentiviral vector GVll8 by T4 DNA ligase and then transformed DH5α competent cells.The plasmids were transformed into the DH5α competent cells.Recombinant colonies were screened by PCR and sequenced.Recombinant plasmids and two other auxiliary plasmids were used to infect 293T cells.Cell culture supernatant was collected for the measurement of viral titer.Recombinant lentiviral vector was used to infect HLE-B3 cells to calculate the viral multiplicity of infection (MOI) under the fluorescence microscope.Transfection efficiency was examined by calculating the GFP-positive cells.HLE-B3 cells were divided into negative control group,siRNA plasmid tranfected group and GV118-Smac-siRNA1 tranfected group.The relative expression levels of Smac mRNA in the cells were detected and compared among the three groups by real-time fluorescent quantitative PCR.Results GV118-Smac-siRNA was successfully constructed with the positive colonies 340 bp and blank vector colonies 299 bp,and viral titer was 3.0× 108 TU/ml.At a MOI of 100,the infecting efficiency of the vector on HLE-B3 cells was about 82％ and the cytotoxicity was low.The relative expression levels of Smac mRNA were (101.290±8.349)％,(92.330±6.320)％ and (32.540±4.221)％ in the negative control group,siRNA plasmid tranfected group and GV118-Smac-siRNA1 tranfected group,respectively,showing a significant difference among the three groups(F =32.871,P＜0.01),and the relative expression level of Smac mRNA was significantly lower in the GV118-Smac-siRNA1 tranfected group than that in the negative control group (P =0.000).However,no significant difference was found in the Smac mRNA expression between the blank plasmid group and the negative control group (P=0.535).Conclusions GV118-Smac-siRNA lentiviral vector is successfully constructed.Smac-siRNA can effectively inhibit the expression of Smac mRNA in human LECs.

To explore the role of connexin43 (Cx43) in gap junctional intercellular communication (GJIC) and propagated sensation along meridians, the expression of Cx43 in the rat epithelial cells and fibroblasts was studied both in vitro and in vivo. With the in vitro study, the rat epithelial cells and fibroblasts were cultured together, and the localization of Cx43 was detected by immunohistochemistry and indirect immunofluorescent cytochemistry and under confocal microscopy. And the expression of Cx43 on the surface of the cells was examined by flow cytometry. With the in vivo examination, 20 SD rats were randomized into control group (n = 10) and electrical acupuncture group (EA group, n = 10). EA ( 0.5-1. 5 V, 4-16 Hz , 30 min) was applied to "Zusanli" acupoint for 30 min at rat's hind paw, the localization of Cx43 was immunohistochemically detected. The immunohistochemical staining and indirect immunfluorescent cytochemistry showed that Cx43 was localized on the surface of the cells and in the cytoplasm. The relative expression level of Cx43 on the cellular membrane surfaces of the rat epithelial cells and fibroblasts, as determined by FACS, were 13.91% and 29.53% respectively. Our studied suggested that Cx43 might be involved in GJIC and propagated sensation along meridians.
Cells, Cultured ; Connexin 43/*biosynthesis ; Connexin 43/genetics ; Epithelial Cells/cytology ; Epithelial Cells/*metabolism ; Fibroblasts/cytology ; Fibroblasts/*metabolism ; Flow Cytometry ; Gap Junctions ; Meridians ; Microscopy, Confocal ; Random Allocation ; Rats, Sprague-Dawley

Objective To investigate the influence of acupuncture on promoting rat blastocyst nidation with Cx43 expression at the acupoints silenced, and to relate Cx43 with acupuncture's effects. Methods Ma-ted female rats were randomly divided into five groups: normal control, model, acupuncture, interference-control and interference. Mifepristone was used to induce a model of nidation disorder, and RNA interference technology was used to silence the expression of Cx43 at acupoints. P-Cx43-shRNA (1) was injected locally into the acu-points of rats in the interference group, and P-con-shRNA was injected in the interference-control group. Acu-puncture was administered to the acupuncture, interference-control and interference groups for 7 days. On the 8th day of pregnancy, the quality of blastocysts, pregnancy rates, and the average amount of blastocysts in each group were observed. Results ①The mRNA and protein level of Cx43 at the zusanli (ST36) acupoints in the interference group were significantly lower than in the acupuncture group ( P<0.05, P<0.01). There was no significant difference between the interference-control group and the acupuncture group. ② Compared with the normal control group, the quality of blastocysts in the model and interference groups was bad, and the amount was less. Their size was comparatively small, they were inhomogeneous, poorly distributed and had poor blood supply. The results improved in the acupuncture group and the interference-control group compared with the model group. ③The pregnancy rates in the normal control, model, acupuncture, interference-control and inter-ference groups were 100% , 50% , 75% , 75% , and 50% respectively, but these differences were not statisti-cally significant. ④The average amount of blastocysts in the model and interference groups was significantly less than in the normal group (P≤0.01). Compared with the acupuncture group, the average amount of blastocysts in the interference group was lower (P≤0.05). Conclusion Acupuncture can improve the quality and quan-tity of blastocysts in a rat model of nidation disorder. Local injection of Cx43 shRNA expression vector can si-lence the expression of Cx43 in acupoints and markedly influence acupuncture's effect. This demonstrated that Cx43 was involved in acupuncture's effects.

Objective To explore the effect of different controlled hypotension method on the blood flow of the vertebral vein by measuring of blood flow of vertebral vein in rabbits. Methods Eighteen New Zealand rabbits,weighing 2-3 kg,were rando mly divided into three groups (n =6 each).Group S maintained MAP (90±5)mm Hg,group GTN reduced MAP to the base value of (70±10)% by using nitroglycerin 0.5 μg·kg-1 ·min-1 and esmolol 2.5 μg·kg-1 ·min-1 (ni-troglycerin∶esmolol= 1∶5 ),group SNP reduced MAP to the base value of (70 ± 10 )% by using sodium nitroprusside 0.5 μg·kg-1 ·min-1 and esmolol 2.5 μg·kg-1 ·min-1 (sodium nitroprusside∶es-molol=1∶5).The controlled hypotension model was established after intravenous general anesthesia. The blood flow of vertebral vein in rabbits were measured by ultrasonic measurement system (Terason 2 000 system ) before controlled hypotension and 1 hour after controlled hypotension. Results The MAP before controlled hypotension in group S (89.0 ± 5.2 )mm Hg,group GTN (91.5± 9.6 )mm Hg,group SNP (92.0 ± 5.7 )mm Hg had no significant difference.The blood pressure before and after the experiment had no significantly difference in group S.The blood pressure after controlled hypotension had no significant difference in group GTN and group SNP,but lower than that before controlled hypotension (P <0.05).Compared with group S,the blood flow of verte-bral vein in group GTN and group SNP were significantly reduced (P < 0.05 ).The blood flow in group GTN was significantly reduced compared with that in group SNP (P <0.05 ).Conclusion In the experiment,the combination of nitroglycerin and esmolol can better reduce blood flow of vertebral vein than that of nitroprusside and esmolol,that it is suitable for the control of hypotension in spinal surgery.

Background Posterior capsular opacification (PCO) is a primary complication after extracapsular cataract extraction.The mechanism of PCO is associated with proliferation,migration and epithelialmesenchymal transition (EMT) of human lens epithelial cells (LECs).To explore the target treatment of PCO is very important.Objective This study was to investigate the biological effects of second mitochondria-derived activator of caspases (Smac) on the proliferation and apoptosis of LECs.Methods Human LECs line (HLE-B3) and Smac-overexpressed LECs line were cultured,and the cells were transfected using small interfering RNA (siRNA)-Smac3 plasmid with green fluorescent protein (GFP) for 24 hours.Different concentration of transforming growth factor-β2 (TGF-β2) (5,10,20 and 50 μg/ml) or 200 μmol/L H2O2 were added respectively into the culture medium to establish PCO model and oxidative stress model.Cell counting kit-8 (CCK-8) assay was used to compare the cell proliferative activity among PBS group,TGF-β2 group and Smac-hyperexpression +TGF-β2 group.Flow cytometry was used to evaluate the apoptotic rate of the PBS group,H2 O2 group and siRNA-Smac+H2 O2 group.The expressions of Smac,caspase-3 and proliferating cell nuclear antigen (PCNA) mRNA and their proteins in the cells were detected by real-time quantitative PCR (RT-PCR) and Western blot.Results The GFP+ cells were≥ 80％ 12 hours after siRNA-Smac3 transfection,with the optimal plasmid of siRNA-Smac3.GFP+ cell rate was (72.32 ± 2.31)％ in the siRNA-Smac3 transfection group,which was significantly higher than that in the blank plasmid group ([4.91 ±0.24] ％) (t=116.342,P＜0.001).The relevant expression levels of Smac was 35.21 ±4.11 in the Smachyperexpression group,and that in the blank plasmid group was 15.24±2.48,with a significant difference between them (t =215.47,P＜0.05).The cell viability of 20 ng/ml TGF-β2 affected PBS group,TGF-β2 group and Smachyperepression+TGF-β2 group was (98.4 ± 1.7) ％,(98.9 ± 0.1) ％ and (64.2 ± 3.1) ％,and the cell viability of Smac-hyperepression+TGF-β2 group was significantly lower in the Smac-hyperepression+TGF-β2 group than that in the TGF-β2 group (P＜0.05).The apoptotic rate in the PBS group,H2 O2 group and siRNA-Smac+H2 O2 group were (2.9 ± 1.2) ％,(45.1 ±4.5) ％ and (27.5 ± 1.8) ％,and the apoptotic rate was evidently lower in the siRNA-Smac +H2O2 group than that in the H2O2 group (P＜0.05).RT-PCR results showed that the expression levels of caspase-3 mRNA in PBS group,H2 O2 group and siRNA-Smac + H2 O2 group were 0.321 ± 0.103,0.715 ± 0.112 and 0.479 ±0.209,respectively.Compared with the H2 O2 group,the relative expression level of caspase-3 mRNA in siRNA-Smac+ H2O2 group was significantly decreased,the difference was statistically significant (P＜ 0.05).The PCNA mRNA expression levels in PBS group,TGF-β2 group and Smac-hyperepression+TGF-β2 group were 0.299±0.013,0.645± 0.102 and 0.490±0.209,respectively.Western blot results showed that the relative expression of caspase-3 protein in siRNA-Smac+H2O2 group and H2O2 group was 0.712±0.012 and 0.973±0.051,with significant difference between the two groups (t =132.52,P＜0.05).The relative expression of PCNA protein in Smac-hyperepression+TGF-β2 group was 0.782±0.212,which was lower than 1.126±0.251 in the TGF-β2 group (P＜0.05).Conclusions Smac may prevent and treat PCO by inhibiting the proliferation and promoting apoptosis of human LECs.

Objective To discuss the characteristics of extended-spectrum beta-lactamases(ESBLs)-producing Shigella and the relation between them and drug-resistance plasmid. Methods The suspicious ESBLs-producing isolates were screened by K-B disc diffusion method, and the ESBLs-producing strains were confirmed by confirmatory test recommended by the National Committee for Clinical Laboratory Standards. Furthermore, the partial blageneof these isolates were detected by PCR using universal primers for TEM, SHV, CTX-M-1 group, CTX-M-2 group and CTX-M-9 group, respectively. The entire blaCTX-M-9 and blaTEM were amplified by PCR using the primers outside the open reading frame (ORF) of these β-1actamases and products were directly sequenced. The conjugation experiment was performed to determine whether the resistance was transferable. Minimal inhibitory concentration (MIC) was detected with double agar dilution method. Results Of the 275 isolates, 12 strains were identified as ESBLs producers. Among them, 8 strains were CTX-M-14 carriers and 4 strains were CTX-M-3 carriers. All ESBLs-producing isolates are positive for plasmid conjugative transfer test. The transconjugants are only resistance to betalactams. Conclusions High resistance to beta-laetams in Shigella is caused by production of ESBLs in the local area. The ESBLs-produeing isolates can transfer the drug resistance through lateral transfer of plasmid.

Objective To compare high-frequency ultrasonography and MRI in diagnosis of medial patellofemoral ligament (MPFL) injury after acute lateral traumatic patellar dislocation(LTPD).Methods According to surgical data of 35 patients with acute LTPD,the incidence and sites of MPFL injury were analyzed.Further,based on high frequency ultrasonography and MRI data,the sensitivity,specificity and accuracy of high-frequency ultrasonography and MRI in different MPFL injuries were analyzed and compared.Results Among 35 patients,there were 21 cases with complete MPFL tear and 14 cases with partial MPFL tear.Tear of MPFL at the femoral attachment in 19 cases,at the patellar attachment in 15 cases,and at the mid-substance in 1 case.The sensitivity,specificity,accuracy of high-frequency ultrasonography diagnosis were 78.6％,95.2％,88.6％ respectively on partial MPFL tear and 95.2％,78.6 ％,88.6 ％ respectively on complete MPFL tear.The sensitivity,specificity,accuracy of MRI diagnosis were 85.7％,90.5％,88.6％ respectively on partial MPFL tear and 90.5％,85.7％,88.6％ respectively on complete MPFL tear.There were no significant difference between high-frequency ultrasonography and MRI in diagnosis of partial and complete MPFL injury (x2 =0.243,0.267; P =0.622,0.599).Conclusions MPFL formoral attachment is most vulnerable to damage,followed by patellar attachment.High-frequency ultrasonography,as well as MRI,can be used clearly to diagnose MPFL injury and accurately classify the damage,and it is also an easy,reliable,rapid method,which can be applied routinely to diagnose MPFL injury after actue LTPD.

Objective To compare and analyze MRI characteristics of medical patellofemoral ligament(MPFL) injury patterns in adolescents and adults after acute lateral patellar dislocation.Methods MR images of 42 adolescents and 45 adults after acute lateral patellar dislocation were retrospectively reviewed.Routine MR scanning was performed in axial,sagittal and coronal planes,including T1 WI,T2 WI and fat saturation sequence.The x2-tests were performed to analyze the MPFL injury patterns between adolescent group and adult group.Results The prevalence rate of MPFL injury was 97.6％ (41/42) in adolescent group after acute lateral patellar dislocation,including 57.1％ (24/42) of complete tear and 40.5％ (17/42) of partial tear.The prevalence rate of MPFL injury was 100.0％ (45/45) in adult group,including 64.4％ (29/45) of complete tear and 35.6％ (16/45) of partial tear.There were no statistical difference between adolescents and adults in prevalence rate of MPFL injury,complete MPFL tear,partial MPFL tear (x2 =1.084,0.486,0.223 ; P ＞ 0.05).The prevalence rate of MPFL injury at femoral attachment,patellar attachment and midsubstance were 31.0％ (13/42),78.6％ (33/42) and 26.2％ (11/42) respectively in adolescent group,whereas they were 64.4％ (29/45),40.0％ (18/45) and 15.6％ (7/45) respectively in adult group.It occurred at multiple sites in 33.3％ (14/42) of adolescents,and in 15.6％ (7/45) of adults.There were significant differences between adolescents and adults in the prevalence rate of MPFL tear at femoral attachments and patellar attachments (x2 =9.759,13.324 ; P ＜ 0.05),but there was no statistical difference at its midsubstance (x2 =1.497,P ＞ 0.05).Although not statistically significant (x2 =3.749,P ＞ 0.05),MPFL tear at multiple locations were seen more in adolescents than in adults (33.3％ vs.15.6％).Conclusion The injury degree and distribution of MPFL are similar in adolescents and adults after acute lateral patellar dislocation,but there is significant difference in the site of MPFL injury distribution between adolescents and adults.MPFL is most easily injured at femoral attachment,secondly at patellar attachment in adults,while MPFL is most easily injured at patellar attachment,secondly at femoral attachment in adolescents.Adolescents are more likely to have multiple-site MPFL injury than adults.

Objective To investigate the role of large-diameter magnetic resonance imaging (MRI) simulation in target volume delineation in radiotherapy for nasopharyngeal carcinoma (NPC).Methods Eighteen patients with NPC underwent computed tomography (CT) simulation and MRI simulation scans and localization in the same body position,with SOMATOM Sensation Open 40-row 82-cm large-diameter CT simulator and Siemens 3T MRI MAGNETOM Skyra 70-cm large-diameter simulator,respectively.The gross tumor volume (GTV) and parotid glands were delineated on all images according to the ICRU Report 50/62,and MRI was applied to observe the changes in GTV and parotid volume during radiotherapy.Paired t-test was applied to analyze the differences between GTVCT and GTVMRI and between GTVnx-CT and GTVnx-MRI.Results GTVMRI decreased significantly compared with GTVCT,and the average volume decreased from (213.64±84.59) cm3 to (199.68±84.69) cm3(p=0.006).As for the volume of primary lesions in the nasopharynx,GTVnx-MRI was significantly smaller than GTVnx-CT,and the volume decreased from (95.75± 24.76) cm3 to (88.12±26.25) cm3 (P =0.001);as for the volume of cervical lymph nodes,GTVnd-MRI was significantly smaller than GTVnd-CT,and the volume decreased from (117.89± 72.69) cm3 to (111.56± 70.69) cm3 (P=0.018).The targets delineated by CT and MRI did not overlap completely,with major differences in skull base bone and cervical soft tissue.The volume of both parotid glands delineated on MRI image was higher than that delineated on CT image,with a major difference in the deep lobe.MRI showed that GTV was reduced by 82.64± 16.87％ during radiotherapy,and the volumes of the left and right parotid glands were reduced by (32.7± 23.95) ％ and (34.7± 21.72) ％,respectively.Conclusions The delineation of target volume based on MRI simulation is more accurate than that based on CT simulation and can achieve a smaller volume range,which helps to guide target volume delineation in radiotherapy for NPC accurately.