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Objective@#To study the difference expression and diagnostic value of ribosomal protein L5 (RPL5) in papillary thyroid carcinoma (PTC) of children and adults.@*Methods@#Realtime-PCR was performed to detect the expression of RPL5 in 22 PTC tissues and 13 pericarcinous tissues. Receiver operating characteristic (ROC) curve and Youden's index were used to evaluate the diagnostic value of RPL5 in PTC of children and adults.@*Results@#The expression of RPL5 in PTC tissues was higher than in pericarcinous tissues. The area under curve (AUC) was 0.820 (P=0.001), and Youden′s index was 0.568. The expression of RPL5 in PTC of adults was higher than children (P<0.05). The AUC and Youden's index were respectively 0.721 (P=0.069) and 0.414 in children, whereas being respectively 0.896 (P=0.0005) and 0.709 in adults. RPL5 in diagnosis of PTC of adults was better than CK19, Galectin-3 and TPO, which are commonly used for the pathologic diagnosis of PTC.@*Conclusion@#The expression of RPL5 in PTC is higher than pericarcinous tissues, and its expression in PTC of adults is higher than children. Furthermore, PTC is a potential indicator for diagnosis of PTC.
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Objective To evaluate the sensitivity, repeatability and accuracy of microarray digital PCR system in detecting JAK2 V617F mutation, which was closely related to myeloproliferative neoplasms (MPN).Methods All of the 31 MPN patients with JAK2 V617F mutation, including 18 cases of polycythemia vera(PVs),11 primary thrombocythemias (ETs) and 2 primary myelofibrosis (PMFs), were collected from Huashan Hospital, Fudan University during 2014 -2015, while 10 normal controls and 6 cases with abnormal increased hemoglobin were involved.Human erythroleukemia cell line ( HEL ) and colorectal cancer cell SW480 were used as the mutant and the wild type control, respectively.The sensitivity of microarray digital PCR were verified by detecting the gradient diluted mutation standard harboring 30%, 10%, 1%, 0.1%and 0.01%mutant allele burden, respectively .Repeatability was evaluated by detecting 1%and 10% mutated samples for 5 times, respectively.MGB probe real time PCR was selected as the reference method to verify the accuracy of the digital PCR.Results With digital PCR, the accurate quantitation of JAK2 V617F mutation was achieved down to 0.1%, which is approximate to 0.16 copies per microliter.The results obtained from the two kinds of technique showed a high correlation by linear regression analysis (R2 =0.998 3).The results of repeated samples showed CVs as 17.18% for 1%mutant allele burden and 7.50%for 10%.Among all cases, the 31 patients known mutated were detected as positive and 10 controls as negative by both digital PCR and Real time PCR.In another 6 cases, 2 were found JAK2 V617F mutation of low allele burdens of 0.37% and 0.18% by digital PCR but detected as negative by real time PCR.Conclusions Microarray digital PCR offers a higher sensitivity and better repeatability than real time PCR which could help detect rare JAK2 V617F mutations in MPNs accurately.
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Objective To discuss the relationship of drip velocity of nitrate on blood pressure while treating coronary disease,in order to provide appropriate drip velocity for clinical treatment.Methods 155 patients with coronary disease using nitrate to lower blood pressure were selected.They were divided into the nitro glycerin group(85 cases) and the isosorbide mononitrate group( 70 cases) according to difference of medication.The velocity of drugs was adjusted on basis of blood pressure changes.The blood pressure changes at different drip velocities were observed and compared.Results The systolic pressure and the diastolic pressure between two groups showed no difference at 20 drops/min,but the results were the opposite at 30 drops/min.The systolic pressure and the diastolic pressure in the nitro glycerin group showed evident changes at different drip velocities,but in the isosorbide mononitrate group,these changes were not so significant.9 patients in the nitro glycerin group had headache during treatment,no headache occurred in the isosorbide mononitrate group.Conclusions Intravenous use of nitrate at a velocity of 20 drops/min is relative secure.The risk of hypotension will increase if the medication speed increases.lsosorbide mononitrate has little influence on blood pressure.
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Objective To observe the effects of nerve impulses on the expression of carbonic anhydrase Ⅲ ( CAⅢ ) and its phosphatase activity, and to explore whether or not the cause of CAⅢ expressive decreased in skeletal muscles of myasthenia gravis( MG) is resulted from the obstruction of nerve impulse.Methods The motor nerves of extensor digitorum longus (EDL, mainly composed by fast fibers) and soleus (Sol, mainly composed by slow fibers) were cut off by operation of denervation.Levels and phosphatase activities of CAⅢ were analyzed at 7, 14, 28, and 56 d after denervation by Western blot and specific enzyme staining on the membrane following SDS-polyacrylamide gel electrophoresis, respectively.Results (1) Levels of CAⅢ in Sol of normal side (eg denervated contralateral) were much higher than that in EDL of normal side, and the levels in both Sol and EDL had an enhanced tendency with time (age) increase, especially for Sol.After denervation, the levels of CAⅢ in EDL were gradual increased, however, the level in Sol was 14 d after denervation as the boundary of ascension and then decline.( 2) The phosphatase activities of CAⅢ in Sol of normal sides were much higher than that in EDL of normal sides, and there were an enhanced tendency with time (age) increase in Sol, but no significant changes were found in EDL The enzyme activities in denervated Sol were lower(in the 14, 28, and 56 days after denervation: 14.39 ±1.93, 11.48 ±1.46, 9.04 ±1.46) much than their contralaterals(22.75 ± 1.80, 25.26 ±3.15, 25.82 ± 2.97; t = 0.002, 0.005, 0.002, all P ＜ 0.05), the enzyme activities in denervated EDL were also lower than their contralaterals, however, no significant differences were found.(3)It was consistent for CAⅢ levels and phosphatase activities in both Sol and EDL of normal sides.After denervation, however, the deviation of the CAⅢ levels and phosphatase activities happened, the levels of CAⅢ were increased, but the phosphatase activities were decreased.Conclusions The effect of nerve impulse transferring obstructed by denervation on CAⅢ expression of skeletal muscles is different from that by MG auto-antibody.The decrease of CAⅢ protein in the MG muscles may be not resulted from the nerve impulse transferring obstructed by MG auto-antibody.
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Objective To evaluate Western blot analysis in diagnosing limb-girdle muscular dystrophy type 2A (LGMD2A). Methods The clinical records including their pathological and biochemical results of 4 patients with LGMD type 2 were reviewed. Histochemical and immunohistochemical staining were performed on muscle biopsy specimens from the four patients. The expressions of dysferlin and calpain-3 in muscles were analyzed by Western biol. Results All 4 LGMD patients shared some common clinical features, such as dorsal muscular atrophy of lower limbs and remarkably elevated CK. The immunohistochemical results showed partial or complete deficiency of dysferlin staining in all 4 LGMD patients. However, Western blot revealed that the calpain-3 protein in the muscle of patient 1 was completely absent, who was later diagnosed with LGMD2A. The other 3 patients had complete dysferlin deficiency with reduced calpain-3 expression and they were confirmed to be LGMD2B. Conclusions Western blot analysis of calpain-3 and dysfcrlin can be used to differentiate LGMD2A which shows absence of calpain-3 from other LGMD types which show dysferlin deficiency. Western blot is an invaluable method in clinical diagnosis of LGMD2A.
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Objective To observe the effects of lentivirus-mediated RNA interference silencing IL-1 type Ⅰ receptor on the expression of mitogen-activated protein kinase (MAPK) in a rat model of osteoarthritis (OA), and to explore the role of IL-1 and MAPK in OA. Methods Forty SD rats were randomly divided into four groups(A-D, each group: n = 10). The rats (A-C group) were underwent unilateral anterior cruciate ligament transaction and partial excision of the medial meniscus. At day 7 and 9 after operation, the rats of group A were respectively received 40 μL of lentivirus-mediated RNAi silencing IL-1 type Ⅰ receptor by intra-articular injection, while the rats of group B were received lentivirus-mediated RNAi non-silencing IL-1 type Ⅰ receptor, and the rats of group C were received saline. The rats of group D were taken for normal control. All rats were sacrificed four weeks after the surgery. The knees were harvested to observe macropathologic changes of the joint cartilage. ERK1/2, JNK1/2 and p38 proteins in the cartilage were detected by Western blot. Results Cartilage degradation in group A was milder than that in group B and C (P<0.05),but was worse compared to group D (P<0.05). Level of p38 expression in cartilage in group A was lower than that in group B and C (P<0. 05), but had no significant difference compared to group D (P>0.05). Levels of ERK1/2 and JNK1/2 expressions in cartilage in group A were lower than that in group B and C(P<0.05),but was significantly increased compared to group D(P<0.05). Conclusions Lentivirus-mediated RNAi silencing IL-1 type Ⅰ receptor can inhibit the expression of MAPK, in particular,p38 protein was strongly inhibited.
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Objective To detect the effect of the antiphospholipid antibodies on the Schwann cells in vitro and to find if the Extract of Ginkgo biloba 761(EGb761)has any protective effects.Methods The immunoglobulins(Ig)were extracted from the serum of the patient with autoimmune polyneuropathies and those with controls.Ig only or Ig plus calf serum(served as complement)were added into the culture solutions of the Schwann cells.The density of the cell in one high power field,the cell perimeter and area were detected and compared to those blank and those controls.Results The shape of the Schwann cells in both the Ig and Ig plus calf serum solution were greatly changed.The densities of the cells in them were both lower than the blank ones,with more severe in the cells with calf serum.There was no significant different between the solutions with or without EGb761.Conclusion Ig from the patients with autoimmune polyneuropathies could attack the Schwann cells in vitro,with the existence of complements,the destruction was even more severe.No protection of EGb761 could be found in these situations.
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BACKGROUND: Since the discovery of the fact that activin can promote the survival of retinal neurocyte in chicken,the effects of activin in nervous system receives recognition. As discovered recently,hippocampal activin βA mRNA expression up-regulates in multiple brain injury animal models including ischemia and hypoxia; however,the change of activin βA mRNA expression after epilepsy is waiting for investigation.OBJECTIVE: To observe hippocampal activin βA mRNA expression at different time point after pilocarpine (PC) -induced epilepsy in mouse to explore its mechanism.DESIGN: A randomized controlled experimental study based on the experimental animals.SETTING: Department of neurology in a university affiliated hospital and the institute of neurology in a university.MATERIALS: The study was conducted in the Institute of Neurology of Huashan Hospital Affiliated to Fudan University and the Department of Pathology of Shanghai Medical College between November 2001 and July 2002. Totally 168 eight to ten-week old healthy male C57BL/6 mice with a body mass between 20 g and 25 g were obtained from Shanghai Experimental Animal Center,Chinese Academy of Science.INTERVENTIONS: 350 mg/kg(10 g/L) of PC was injected into the abdominal cavity in the mice of study group,in which 1 mg/kg of scopolamine (SC) was injected at 30 minutes before the injection of PC to antagonize its peripheral cholinergic reaction. Status epilepticus(SE) model mouse was the mouse with continuous mgoelonus or generalized seizure of rigid clonus that lasted for 1 hour after the injection of PC. Valium(4 mg/kg) was immediately injected after the modeling to terminate seizure. Same dose of Valium was injected into non-SE(NSE) mice after 1.5 hours of PC injection. Saline was used to replace PC to inject into mice of control group,and the rest disposals of control group were as same as that of study group. SE mice,NSE mice and control mice were randomly divided into six subgroups including 0hour,1 hour,3 hours,6 hours,24 hours and 48 hours subgroups according to the time point after modeling with 6 mice of each subgroup(mice of NSE group and subgroups of 0 hour time point were not included into analysis of hybridization in situ).MAIN OUTCOME MEASURES: Hippocampal activin βA mRNA expression of different time point in SE mice and NSE mice were observed by RT-PCR; the distribution of hippocampal activin βA mRNA expression at different time points in mice were observed by hybridization in situ.RESULTS: There was no significant change of hippocampal activin βA mRNA expression at different time point in mice of NSE group and control group. In SE group,activin βA mRNA(0.49 ± 0. 11) had a transient significant decrease at the beginning(0 hour),which rapidly returned to control level(0. 74 ±0. 13) at 1 hour(0.73 ±0. 12) . Activin βA mRNA continuously increased and reached (0.97 ±0. 24) at 3 hours,(1.34 ±0. 19) at 6 hours,maintained (0.98 ±0. 17) until 24 hours,and decreased to (0. 83 ± 0.09) at 48 hours afterwards,which was slightly higher than control level. Compared with control group,the increases at 3 hours,6 hours and 24hours were significant( t = 2. 668,6. 289,2. 916,P ＜ 0. 001 - 0. 05). The significant up-regulation of activin βA mRNA expression was occurred earliest in hippocampal CA2 and DG regions at 3 hours after SE,and the significant expressions also could be seen in CA3 region after 6 hours. There were expressions in only CA2 and CA3 regions after 24 hours,while there were very few positive cells in CA2 region after 48 hours.CONCLUSION: PC-induced SE could significantly up-regulate hippocampal activin βA mRNA expression,while NSE has no such up-regulative effect. The up-regulation of hippocampal activin βA mRNA expression might be an endogenous protective effect of neuron on antagonizing excitatory injury.
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Oblective To evaluate the corneal healing of non-epuality diopter response histopathologically,immunohistochemically and ultrastructurally after excimer laser photorefractive keratectomy(PRK) with SVS APEX PLUS(Summit Technology Inc. USA) excimer laser, and the effects of corticosteroid on the healing. Methods PRK on 6 white rabbits(12 eyes) was performed on right eye of the rabbit for an attampted correction of -4.00 diopter and on left eye for an attempted correction of -8.00 diopter. The rabbits were divided into 2 groups randomly and each group included 6 eyes: Group FLM (3 rabbits, 6 eyes) and group CM (3 rabbits, 6 eyes). Fluoromethalone was given to group FLM,and chloromycetin to group CM. On 10d, 30d and 100d ,the eyes of one rabbit in each group were enucleated randomly. Half of each cornea was prepared for electron microscope observation (SEM and TEM)and the rest embedded in OCT compound for immunohistochemical study to examine Ⅲ -C and FN. Results All eyes were reepithelialized within 3d after PRK. Subepithelial corneal haze was observed on 15d,which was dominant on 30 or 60d. On 100d postoperatively,corneal hazes of 11 eyes were grades 0 or 0. 5,only 1 eye(the left eye of group CM) was denser haze (grade 1). On 3d postoperatively, one or two layers of corneal epithelial cell covered the ablation zone. On 30d after PRK,the epithelial cells showed hyperplastic changes. The cells were larger and increased from normal 5 or 6 layers to 7 or 8 layers of cells on 100d after PRK,epithelium was clear with more bright epithelium. Microplicae and microvilli were less than before. The expression of Ⅲ -C and FN in group CM was significantly more evident than that in group FLM. Conclusion The study show that despite recovery of a continuous and smooth epithelial layer and nearly normal corneal tissues on 100d after PRK,abnormalities of both epithelium and superficial stroma can be detected in the area of ablation. The ablation depth of stroma can influence the appearance of corneal haze after PRK. The microplicae and microvilli of rabbit cornea epithelium become less after PRK,which can be one of cause leading to ocular dry sensation in some patients.
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Objective Expressing the human matured brain-derived neurotrophic factor (mBDNF) gene in E.Coli and determining its bioactivity. Methods The resulting gene of mBDNF was subcloned into the EcoRI-BamHI site of the expression vector plasmid pBV220. The ligation products were used to transform the competent E. Coli DH5α. The proteins of mBDNF were experessed by temperature inducing. The expression products were dealed with solubilizing inclusion bodies and refolding protein. It was introduced into the embryonic chicken DRG to test whether the expressed mBDNF is a biologically active protein. Results The recombinant plasmid pBV/mBDNF was successfully constructed. By temperature inducing,under the control of the bacteriophage λ PL promoter, the experessed mBDNF protein was a 14Kd non-fusion protein,which existed in E. Coli as inclusion bodies. The size of expressed mBDNF is identical to the prediction. Bioactivity of the products was proved that it could support the cell survival and neurite growth in the primary cultures of embryonic 8-day-old chicken DRG neurons as compared to control.Conclusion The mBDNF gene can be expressed bioactively in E. Coli.
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Objective To investigate integration and expression of adeno-associated virus (AAV) vectors in neuronal PC12 cells,the result of which can be applied in further gene therapy of diseases of the central nervous system. Methods Human neurotrophin-3(hNT3)genes were inserted into AAV vectors. Then the recombinat AAV plasmids were encapsidated as recombinant virions. PC12 cells were transfected with the virions and the positive cells were selected by G418. The transfection positive (hNT3 modified)PC12 cells were cultured for several generations and the cellular genomic DNA and total RNA were extracted. We investigated the integration locus of AAV vectors by Southern blot and transcript situation of foreign genes by dot blot. Results The hybridization tests showed that AAV introduced foreign genes were stably integrated, but at random locus, and robustly transcribed in hNT3 modified PC12cells. Conclusion AAV vectors can serve as high efficiency vectors of target genes in neuronal PC12 cells.
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Objective To study the effect of ginsenoside Rb1(Rb1) and total saponin of dipsacus asper(tSDA) on intracellular free calcium concentration([Ca2+]i) mediated by β-amyloid protein(A β).So as to lay a foundation for developing effective Chinese traditional medicine to treat Alzheimer's disease(AD).Methods The technique of laser scanning confocal microscopy combining primary cultured neurons was adopted to quantitatively analyze the change of [Ca2+]i.Results The [Ca2+]i of primary cultured hippocampal neurons was (185.76±56.22)nmol*L-1 on basal levels.Control group showed obvious change of calcium vibration,[Ca2+]i was elevated to (1383.78±62.83)nmol*L-1.The peak of [Ca2+]i of Rb1 group reached (311.95±32.67)nmol*L-1 and was lower than that of control group (P＜0.01).The tSDA group displayed distinct change of calcium vibration too,and [Ca2+]i reached (358.01±35.42)nmol*L-1.There was a significant difference in [Ca2+]i between control and tSDA group (P＜0.01).Conclusion The research indicated that one of mechanisms by which Rb1 and tSDA protected the neurons was to maintain the balance of [Ca2+]i.
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Objective Cloning and sequencing of the human neurotrophin-4(hNT-4) gene.Methods With the chromosomal DNA of human blood lymphocytes as template,hNT-4 coding genes were amplified by polymerase chain reaction(PCR) and recombinated into phage vector pGEM-T Easy,which were sequenced by using Sanger's single stranded DNA terminal termination method.Results The sequence of the cloned gene is completely the same as that reported in the literature(GenBank data base,M86528).Conclusion This study successfully cloning and sequenced the gene of mhNT-4,and it would be convenient for us to study the expression of mhNT-4 in eukaryote,and to continue the research on the gene therapy of Alzheimer's disease intensively.This study indicate that the hNT-4 is conservative in different races and individuals.
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Objective To explore whether the 25 kD protein component was a specific decrease in skeletal muscles of the patients with myasthenia gravis (MG). Methods The muscular proteins were extracted from 21 cases of normal objects,18 cases of patients with myasthenia gravis and 20 cases of patients with other neuro-muscular disorders with Guba-Straub solution. The components of protein were analysed by SDS-PAGE in double blind. Components of glycoprotein were detected by using the ConA/HRP.Results SDS-PAGE patterns showed that the concentration of protein bands with mass of about 25 kD in the MG muscles was much lower than that of muscles in both normal and other neuro-musclular disorders. The value of density for 25 kD protein bands was 1.6?0.7 in the MG muscles,but was 3.4?1.5 and 3.7?1.5 in the muscles of normal and other neuro-musclular disorders,respectively. In addition,25 kD protein was found as a glycoprotein,but it was different from AChR in the molecular weight.Conclusion (1) It was suggested that the pathogeny or developing of MG could be associated with 25 kD protein of the skeletal muscle because of its specific decrease. (2) 25 kD protein was a glycoprotein which was unassociated with the AChR molecule.
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Objective To explore whether the dopamine D1 and D2 receptors were involved in pathogenic mechanism of Parkinsonian mice induced by paraquat. Methods The models of Parkinson's mice were induced by oral paraquat. The levels of dopamine D1 and D2 receptor proteins and the expression of receptor mRNAs in striatum were examined by immunohistochemistry and in situ hybridization, respectively. Results Mice treated by oral paraquat (10mg?day -1 ?kg -1 ) for four months displayed marked hypoactive behavior. The levels of dopamine D1 and D2 receptor proteins in the striatum were significantly decreased by 28% and 29%, respectively (P
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Purpose To investigate the influence of paraquat on substantial nigra and doparine levels ofstriatum in C57BL mice. Methods 39 neonatal C57BL mice were randomly divided into 5 groups andwere given paraquat or 1-methyl-4-phenyl-1,2,3,6-tetra-hydropyridine(MPTP) orally in 10 th and 11 thdays odl; ( 1 ) MPTP 0.3 mg/kg, n = 8; (2) MPTP 20 mg/kg, n = 8; (3) paraquat 0.07 mg/kg, n = 8; ( 4 )paraquat 0.36 mg/kg, n = 8; (5) normal saline, n = 7. Adult spontaneous motor activity was observed atages of 120 days, then the mice were decapitated and the contents of dopamine(DA), serotonin(5-HT), andtheir metabolites in striatum were analyzed. Meanwhile, the dopamine neuons at the mesencephalon vereobserved by the method of ABC immunohistochemistry. Results Mice given Paraquat 0.36 mg/kg andMPTP 20 rng/kg showed a marked bypoactive behavior and reduced the striatal contents of DA andmetabolites without affecting 5-HT. Immunohistochemical staining showed that the amount of dopamineneurons at the midbrain decreased. Conclusions C57BL mice exposed to great amount of paraquat duringthe neonatal period could yield the alterations of behavior and some pathological and biochemical changessimilar to parkinson disease.
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Objective To define the range of the normal values of MRI signals of hippocampal formation(HPF) for the diagnosis of early stage Alzheimer's disease and hippocampal sclerosis.Methods MRI signals of 254 normal adults were measured on the transverse section.Results HPF signal intensities were:T1 relaxation times,629±73 ms;T2 relaxation times,83±5.5 ms;and proton density value,5978±651.The T2 value range(χ±2S)was 72～94 ms,and mean 83 ms.T2 value greater than 104 ms will be associated with evidence of hippocampal pathologic changes.The mixed signal intensity of T2WI(30,60,120)were:3907±407,2657±347,1288±174.The signal intensity ratios of T2WI were:1.02,1.07,1.13.Conclusion As for the histological features of temporal lobe cortex and HPF,T1relaxation times and proton density values are matched but T2 relaxation times of HPF are relatively longer.
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AIM: To evaluate the effect of carbon monoxide(CO) on the permeability of brain blood barrier(BBB) in cerebral ischemic rats. METHODS: SD rats were divided into three groups. Saline, hemin or ZnPP were injected intraperitoneally 12 h before middle cerebral artery occlusion (MCAO), respectively. The concentration of blood CO and the permeability of BBB at 24 h after MCAO were measured. RESULTS: The CO concentration in blood in hemin group was higher than that in saline group( P 0.05). CONCLUSION: CO reduced the permeability of BBB as a messenger gas molecular when its intrinsic concentration was elevated.
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Objective To evaluate the efficacy and accuracy of photorefractive keratectomy(PRK) for myopia and myopic astigmatism.Methods SVS APEX PLUS excimer laser with a wave length of 193nm(Summit Technology Inc.Waltham,Mass,USA) was applied.Three hundred and sixteen myopic eyes of 168 patients were treated with PRK from Suptember 1996 to October 1997,and 260 eyes(84%) of 150 patients were followed-up for more than three month,including male 68(116 eyes,40.1%) and female 82(144 eyes,59.9%).The preoperative spherical equivalent refractive errors ranged from -1.25D to -10.00D(mean -4.67D± 1.63D),and astigmatism ranged from 0 to -2.00D(mean -0.33D±0.45D). The patients were divided into two groups according to the refraction:group A(from -1.25D to -5.90D) and group B(from -6.00D to -10.00D).The number of eyes in the two groups were 220 and 40,respectively.Results In group A,After 10 days,68.9% had the uncorrected visual acuity(UCVA) equal to or one line better or lower than the preoperative best corrected visual acuity(BCVA).After 1 ,3,6 and 12 months,90%,96%,95% and 94% had the UCVA equal to or one line better or lower than the preoperative BCVA,respectively.In group B,after 10 days,1,3,6 and 12 months,UCVA equal to or one line better or lower than the preoperative BCVA occurred in 35.9%,83.0%,87.0%,86.0% and 84.0% of the patients respectively. Most of the haze showed 0.5～1 grades except 3 eyes with the haze of 2 grade at 3 or 6 months and it changed to l and 0.5 grade respectively at one year.after 10 days and 1,3,6,12 months postoperatively,the corneal haze was noted in 32.9%,84.8%,62.8%,9.0%and 2.8% of the treated eyes respectively.Conclusion We found that the 193nm excimer laser PRK is a predictable,safe,stable,and effective refractive surgery for correcting myopia up to-10.00D in Chinese patients,and the effect is better in myopia lower than -6.00D.
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In the present study, we have cloned the gene of human neurotrophin-3 (hNT-3) from the genomic DNA of white blood cells (WBC) by polymerase chain reaction (PCR). The amplification products were cloned into pUC19 and sequenced. Genomic sequence comparison of the cloned fragment and the reported hNT-3 (GenBank M61180) reveals 7 base differences: 1 in the signal peptide, 3 in the prepro peptide, and 3 in the mature hNT-3. Except the 2 varied bases (16th, T to G; 285th, A to C) in the signal peptide and pro-sequence resulted in the change of their encoded amino-acids (Tyr→Asp; Gln→His), the other varied bases have no influence on their respective encoded amino-acids, and all the changes have no influence on the open reading frame (ORF) of the hNT-3.