BackgroundHypoxia-inducible factor-1 α (HIF-1α) specific double-stranded RNA ( dsRNA ) mediated by liposome inhibit reinal neovascularization in mice at dose-dependent manner. ObjectiveThe present study was to investigate the inhibitory effect of dsRNA targeting HIF-1α on retinal neovascularization in mice.MethodsModels of oxygen-induced retinal neovascularization were set up in C57BL/6J mouse through exposure of postnatal day 7 ( P7 ) to (75±3) ％ oxygen for 5 days.Fluorescein conjugated Dextran angiography of retinal vascular was performed to identify the retinal neovascularization.The 8 mice of the normal group were raised in the room air.Fifty-one P7 mice exposed to(75±3)％ oxygen for 5 days and then returned to the room air and assigned to control group ( 3 mice),empty vector group( 3 mice) and gene therapy group (45 mice),and the latter were medially divided to 9 groups randomly according to dose-ratio ( liposomes ∶ plasmid).The pSilencer 2.1-U6 hygro was injected in the model mice of empty vector group,and different dose-ratios of pSilencer2.1-U6 hygro-HIF-1α dsRNA were injected respectively in the model mice of various gene therapy groups.Fluorescein conjugated Dextran angiography of retinal vascular was performed to observe the morphology of new blood vessels,and retinal slides were prepared to score the numbers of nuclei extending beyond the inner limiting membrane( ILM ),and expression of vascular endothelial growth factor(VEGF) was detected in the retina by immunohistochemistry.Results The retinal blood vessels of the normal group formed a fined radial branching pattern.The retinal vascular patterns in the control group and the empty vector group were characterized by decreased central perfusion in both the superficial and the deep layers.The abundant vessels were distorted and irregular in the control group and empty vector group,and the obstructed capillary and lots of neovascular tufts were seen.The retinal neovascularization and non-perfusion distraction in the every gene therapy group were reduced markedly with the most severe appearance in 1 ∶ 1 ( liposomes ∶ plasmid) dose-ratio group.Few vascular endothelial cell nucleus extending beyond the ILM were found in the normal group;while a large number of vascular endothelial cell nucleus were showed in the control group and empty vector group with the occurring rate 100％.Statistically,no significant difference was seen in the number of nuclei extending beyond the ILM between the control group and the empty vector group(11.57±5.85 vs 11.53±6.15),however,that in 1∶1 (liposomes∶plasmid) group was reduced markedly ( 2.17 ± 4.23 ) ( P ＜ 0.01 ).Immunohistochemistry revealed that VEGF was faintly expressed in the normal group but strongly expressed in the control group and the gene therapy group.VEGF expressions of various gene therapy groups were weaker than ones of the control group and the empty vector group.ConclusionsRetinal neovascularization can be efficiently inhibited by intravitreal injection of the pSilencer2.1-U6 hygro-HIF-1α dsRNA mediated by liposome.Proportion of 1 ∶ 1 (liposomes ∶ plasmid)has a maximized efficiency.

AlM:To evaluate the expression of transcriptional factor lslet-1 in retina in experimental retinal neovascularization induced by oxygen.
METHODS: The murine retinal neovascularization were induced by hyperoxia exposure. The morphological observation of retinal neovascularization was performed using angiography by fluorescein dextran injection under the fluorescence microscope, and the new blood vessels were quantified after 5d in room air (17-day-old) by counting the vascular epithelial cell nuclei protruding into viteous cavity using HE stain. Realtime PCR and Western blot were used to examine retinal lslet-1 level in postnatal 7,12, 14,17 and 26d respectively.
RESULTS: A lots of new blood vessels were demonstrated in the mouse retina in hyperoxic group by fluorescein angiography and histological method. Moreover, no significant difference was found in retinal lslet-1 level in postnatal 7d between hyperoxic group and control group, but was significantly higher in postnatal 12, 14 and 17d mice compared with control mice. However, mice at postnatal 26d, expression of lslet-1 in retina decreased to normal level.
CONCLUSlON: ln processing mouse model of retinal neovascularization, sustained hypoxia retinal tissue induce retinal neovascularization by increas the expression of transcription factor lslet-1.

Objective To investigate the expression of Mta-1 in bladder transitional cell carcinoma (BTCC)and to analyze its correlation with the clinical staging,pathologic grading,metastasis and recur- rence,and to explore the possible molecular mechanisms.Methods Samples of 42 cases of BTCC and 12 normal bladder mueosa tissues were examined with immunohistochemical analysis for the expression of Mta- 1,ER,u-PA and PAI-1.Endothelial cells were stained by anti-CD34,and microvascular density(MVD)of carcinoma tissue was calculated.The correlation of Mta-1 expression with the invasion,metastasis,angiogene- sis and recurrence of BTCCs was analyzed;and the correlation of Mta-1 expression with ER,u-PA,PAI-1,and MVD was also analyzed.Results The positive rate of Mta-1 expression in BTCCs was 73.8%(31/42) , while it was 0.0% in normal bladder mucosa tissues(P＜0.01).The expression level of Mta-l increased with the higher clinical stages and pathologic grades of BTCCs;it was higher in recurrence group(100.0% , 15/15)than in non-recurrence group(59.3%,16/27),and high in metastasis group(100.0%,14/14) than in non-metastasis group(60.7%,17/28)(P＜0.05).The expression level of ER increased with the lower clinical stages and pathologic grades of BTCCs;the positive rate of ER expression was 0.0% in 14 ca- ses with metastasis and was 53.6% in 13 of 28 cases without metastasis(P＜0.05);and the rate was 6.7% in 1 of 15 cases with recurrence and 44.4% in 12 of 27 cases without recurrence(P＜0.05).Negative cor- relation was found between Mta-1 and ER expression(r=-0.739,P＜0.01).The positive rate of u-PA ex- pression(59.5%,25/42)was significantly higher in BTCCs than that in normal bladder mucosa tissues (16.7%,2/12)(P＜0.05).Positive correlation was found between u-PA and Mta-1 expression(r= 0.875),while negative correlation was found between u-PA and PAI-1 expression(r=-0.535).The posi- tive rate of PAI-1 expression in normal bladder mucosa tissues(50.0%,6/12)was significantly higher than that in BTCCs(19.0%,8/42)(P＜0.05).In addition,negative correlation was found between PAI-1 and Mta-1 expression(r=-0.706).And positive correlation was found between MVD in BTCCs marked by an- ti-CD34 and Mta-1 expression(r=0.683).Conclusions Mta-1 is highly expressed in BTCCs,and it correlates closely with tumor pathologic grades,clinical stages,recurrence and metastasis.Mta-1 up-regulates the expression of u-PA and down-regulates that of PAI-1,which is associated with invasion and metastasis and acts as an angiogenic mediator in BTCCs.A negative correlation is found between Mta-1 and ER in inva- sion and metastasis of BTCCs.

OBJECTIVETo investigate the features of chronic prostatitis with non-neurogenic detrusor sphincter dyssynergia (NNDSD) and the effects of pelvic floor biofeedback in the treatment of the disease.

METHODSWe included in this study 113 male patients, aged 15 - 48 (mean 36) years and diagnosed as having chronic prostatitis for 1 -2 (mean 1.2) years based on such typical symptoms as frequent micturition, urgent micturition, voiding pain, difficult void, etc, that lasted over 3 months, and the score > or = 1 on the first and second parts of NIH-CPSI. Urethritis, interstitial cystitis, urethral stricture and neurogenic bladder were excluded. All the patients underwent urodynamic examinations for the uroflow curve, Q(max), Pdet. max and MUCP. Biofeedback was carried out for those with non-neurogenic detrusor sphincter dyssynergia, and the effects were evaluated at 10 weeks.

RESULTSTwenty-one (18.6%) of the 113 cases were found to be NNDSD. Biofeedback treatment achieved obvious decreases in Q(max) (8.2 +/- 4.1), Pdet. max (125.1 +/- 75.3), MUP (124.3 +/- 23.3) and MUCP (101.5 +/- 43.6), as compared with 15.1 +/- 7.3, 86.3 +/- 54.2, 65.4 +/- 23.0 and 43.5 +/- 16.7 before the treatment (P < 0.05). Statistically significant differences were observed between pre- and post-treatment scores on voiding pain (4.0 +/- 2.0 vs 2.2 +/- 1.7), urination (7.9 +/- 2.1 vs 2.2 +/- 1.9), life impact (9.6 +/- 2.7 vs 2.9 +/- 2.6) and total scores (21.7 +/- 4.8 vs 8.4 +/- 4.6) (P < 0.05).

CONCLUSIONChronic prostatitis patients with LUTS may have NNDSD, which is urodynamically characterized by low Q(max), high intra-bladder pressure and increased urethral pressure in some patients. Urodynamic examinations may contribute to definite diagnosis and appropriate choice of treatment. Pelvic floor biofeedback has satisfactory short-term effects in the treatment of these patients.

Adolescent ; Adult ; Ataxia ; complications ; diagnosis ; therapy ; Biofeedback, Psychology ; Chronic Disease ; Humans ; Male ; Middle Aged ; Prostatitis ; complications ; diagnosis ; therapy ; Urinary Bladder Diseases ; complications ; diagnosis ; therapy ; Urodynamics ; Young Adult

Rabies virus ; physiology

OBJECTIVETo investigate the clinical characteristics, diagnostic methods and minimally invasive treatment of prostatic utricle cyst.

METHODSWe retrospectively analyzed the clinical data of 9 cases of prostatic utricle cyst, of whom 5 presented with frequent or urgent micturition, 3 with difficult urination or thinning urinary stream, and the other 1 with hemospermia. All the cases underwent ultrasonography and MRI. Transurethral cyst deroofing was performed for 3 of the patients with smaller cysts close to the prostatic urethra, and laparoscopic excision of the prostatic utricle was conducted for the other 6 with bigger cysts behind the prostatic urethra.

RESULTSThe duration of transurethral cyst deroofing ranged from 30 to 50 min and intraoperative bleeding was 20 -70 ml; the mean time of laparoscopic excision of the prostatic utricle was 100 - 150 min and intraoperative bleeding was 30 -50 ml. All the patients were followed up for 3 - 12 months, which revealed normal penile erection and ejaculation, and no urinary tract irritation or difficult urination.

CONCLUSIONUltrasonography and MRI are excellent imaging modalities for accurate depiction of prostatic utricle cyst. Transurethral cyst deroofing is valuable for prostatic utricle cyst close to the prostatic urethra. Laparoscopic excision of the prostatic utricle, owing to its safety, effectiveness, minimal invasiveness, fewer complications and rapid recovery, can be used as the first option for the treatment of prostatic utricle cyst.

Adult ; Cysts ; diagnosis ; surgery ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; Prostatic Diseases ; diagnosis ; surgery ; Retrospective Studies ; Ultrasonography

OBJECTIVETo study the inhibition effect of HIF-1α specific siRNA expression vector pSUPERH1-siHIF-1α on retinal neovascularization in a mouse model of retinopathy of prematurity (ROP).

METHODSThe mouse model of ROP was prepared by the method Smith described. Forty-eight ROP mice were randomly divided into two groups: an experimental group that was intravitreously injected with pSUPERH1-siHIF-1α and a control group that was injected with pSUPER retro vector. The levels of HIF-1α and vascular endothelia growth factor (VEGF) in the retina were examined by Western blot. The retinal neovascularization was evaluated by angiography using FITC Dextran and quantitated histologically.

RESULTSThe levels of HIF-1α and VEGF in the retina in the experimental group were reduced 90% and 65% respectively compared with those in the control group. Meanwhile, the number of retinal neovascular endothelial nucleus outbreaking the inner limit membrane in the experimental group was significantly reduced compared with that in the control group.

CONCLUSIONSThe development of retinal neovascularization of ROP can be markedly inhibited by RNA interference targeting HIF-1α.

Animals ; Disease Models, Animal ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; analysis ; antagonists & inhibitors ; genetics ; Infant, Newborn ; Mice ; Mice, Inbred C57BL ; RNA, Small Interfering ; genetics ; Retinal Neovascularization ; prevention & control ; Retinopathy of Prematurity ; therapy ; Vascular Endothelial Growth Factor A ; analysis

The paradigm of a real world study has become the frontiers of clinical researches, especially in the field of Chinese medicine, all over the world in recent years. In this paper, ethical issues which probably exist in real-world studies are raised and reviewed. Moreover, some preliminary solutions to these issues such as protecting subjects during the process of real-world studies and performing ethical review are raised based on recent years' practices to enhance the scientificity and ethical level of real-world studies.
Biomedical Research ; ethics ; methods ; Humans

Objective To explore the influence of drug dosage, solvent and other main influencing factors on the successful establishment of alloxan-induced hyperglycemia mouse model and the effect on the stability of this model. Methods 160 6-8-week-old Kunming mice ofSPF grade, (male:female=1:1) were used in this study.The influences of different dosages of alloxan and solvent combinations on the successful establishment rate of the model, survival rate, body weight, fasting blood glucose, blood glucose area under curve, serum insulin level and their stabilities were dynamically observed for six weeks.Results By single intraperitoneal injection of 160 mg/kg bw alloxan ( pH 4.5 citrate sodium as solvent) , we were able to obtain a stable experimental hyperglycemic mouse model with higher levels of successful establishment rate (70%), survival rate (75%), fasting blood glucose (15-20 mmol/L), glucose area under the curve (55-65 mmol/L) and a lower but not loss of serum insulin levels (21 mIU/L).Conclusions In the present study we have carefully considered the influence of main factors such as drug dosages, solvent, etc., on the alloxan-induced experimental hyperglycemic mouse model, and successfully established this model after 6-week period observation of its stability.This model may provide a useful tool in the research of experimental diabetes and hypoglycemic functional studies.

OBJECTIVETo analyze the genetic characteristics of two rabies virus isolates from Henan province and to compare their relations hip with known rabies virus isolates and vaccine strains.

METHODSRabies viral antigens were detected in 100 canine brains by immunofluorescence assay method. Rabies virus was isolated through inoculating the suspensions of positive brains into suckling mice. N gene and G were amplified by RT-PCR and sequenced. Phylogenetic trees were constructed for the analysis on genetic characteristics of rabies virus.

RESULTSTwo rabies virus strains were isolated (Henan Hb1 and Henan Sq1). Data from sequential comparison revealed that the nucleotide and amino acid identities of N and G gene between the two isolates were 99.3% and 98.9%, and 98.7% and 98.4% respectively. The two isolates were more closely related to CTN, with the homogeny of N gene and G gene as 89.1% and 85.6%-85.7% at the nucleotide level, but 97.6%-98.0% and 92.3% at the amino acid level respectively, than to other vaccine strains. When comparing with other known viruses including Chinese isolates, the two stains shared closer identity with the isolates from Indonesia, and the rates of homogeny of N and G gene were 92.1%-93.2% and 91.9%-92.1% at the nucleotide level, 97.5%-98.6% and 96.0%-96.2% at the amino acid level, respectively. Data from the deduced amino acid sequences revealed that some amino acid residues including the residues in the N and G antigenic sites were substituted in the two isolates. Furthermore, phylogenetic analysis showed that the two isolates were also more closely related to the strains from Indonesia and vaccine strain CTN than to any other known street viruses and vaccine strains.

CONCLUSIONBoth Henan Hbl and Henan Sql belonged to genotype 1. However, the N and G gene diverged from known street viruses and vaccine strains at either nucleotide level or amino acid level.

Amino Acid Sequence ; Animals ; Base Sequence ; Brain ; virology ; China ; DNA, Viral ; analysis ; Dogs ; virology ; Genes, Viral ; Genotype ; Mice ; Phylogeny ; Rabies Vaccines ; Rabies virus ; classification ; genetics ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA