AIM: To investigate hemodynamic alterations of retrobulbar vessels in proliferative diabetic retinopathy ( PDR) patients with anterior segment neovascularization, before and 3mo after vitrectomy combined with panretinal photocoagulation and to explore the clinical significance.
●METHODS: Color Doppler flow imaging ( CDFl ) was used for measurement of blood flow velocities and resistive indexes ( Rl ) of the ophthalmic artery ( OA ) , short posterior ciliary arteries ( sPCA ) and central retinal artery ( CRA ) in 21 eyes of 21 PDR patients with anterior segment neovascularization. CDFl parameters were obtained before and 3mo after vitrectomy combined with panretinal photocoagulation ( PRP) .
● RESULTS: Peak systolic velocity ( PSV ) and end diastolic velocity ( EVD ) of CRA were significantly increased after surgeries, Rl were decreased significantly (P<0. 05). Parameters of sPCA and OA have no change after surgeries (P>0. 05).
●CONCLUSION: Vitrectomy combined with panretinal photocoagulation might increase the velocity of CRA, decrease Rl and improve ocular blood supply postoperatively. lt may delay or prevent the process of neovascular glaucoma.

Hemorrhoids ; Humans ; Male ; Mucous Membrane

OBJECTIVETo investigate the expression and significance of Cyclin D1 in oral squamous cell ma (OSCC).

METHODSA immunohistochemistry method, Envosion, was employed to test the manifesting Cyclin D1 in pathological slices of 50 OSCC cases and 10 normal cases, and the results was treated with statistical lysis.

RESULTSIn 50 OSCC cases, Cyclin D1 mainly manifested in karyon, and a little in cytoplasm. manifesting rates of Cyclin D1 in the samples was 80.0%, which was significantly higher than the manifesting of 20.0% in normal oral mucous membrane (P < 0.01). The manifestation of Cyclin D1 was correlated with rent pathological grades, clinical phases and lymph node metastasis (P < 0.05).

CONCLUSIONThe abnormal tation of Cyclin D1 is closely related with the occurrence and development of OSCC. Therefore, it can subsidiary index for OSCC treatment and prognosis.

Carcinoma, Squamous Cell ; Cyclin D1 ; Female ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Mouth Mucosa ; Mouth Neoplasms ; Prognosis

OBJECTIVETo explore the methylation status in promoter region of norepinephrine transporter gene (NET, SLC6A2) in heart failure ( HF) patients and its correlation with qi deficiency/blood stasis syndrome (QDS/BSS).

METHODSThirty-six patients with heart failure (NYHA classification III to IV) were recruited in the study (as the heart failure group) and their scores of QDS/BSS were evaluated. Besides, a healthy elderly group (30 cases) and a healthy youth group (30 cases) were also set up. They were recruited from Physical Examination Center of Fujian Provincial Hospital. Pyrosequencing was applied to detect the methylation in promoter region of SLC6A2 gene, and the total methylation index (MTI) of CpG island was calculated. The correlation between the methylation status in promoter region of SLC6A2 and scores of QDS/BSS was assessed using Pearson and Partial analyses. Risk factors were screened and adjusted using Logistic regression.

RESULTSBy one-factor analysis of variance, the total MTI in the HF group (219.72% ± 54.03%) was obviously higher than that in the healthy elderly group (194.47% ± 34.92%) and the healthy youth group (161.60% ± 41.11%) (all P < 0.05). Meanwhile, the total MTI was higher in the healthy elderly group than in the healthy youth group (P < 0.01). By covariance analysis , after controlling age and BMI, the total MTI was higher in the HF group than in the healthy elderly group (P = 0.041), while it was higher in the healthy elderly group than in the healthy youth group (P = 0.016). Age was found to play an essential role in affecting MTI of SLC6A2 gene promoter region among the 3 groups (F = 16.447, P = 0.01). The total MTI was quite lower in the healthy youth group. Results of Partial correlation analysis showed MTI was positively correlated with scores of qi deficiency and blood stasis respectively (r = 0.494 and 0.419 respectively, both P < 0.05). Logistic regression analysis showed after adjusting confounding factors, the relative risk (OR value) of total MTI of SLC6A2 gene in promoter region was 1.038 (95% CI, 1.006 to 1.071, P = 0.020).

CONCLUSIONSAbnormally elevated methylation of the promoter region of SLC6A2 gene is one of risk factors for HF. In addition, the degree of methylation of the promoter region of SLC6A2 gene was positively correlated with the severity of QDS/BSS.

Adolescent ; Aged ; DNA Methylation ; Heart Failure ; genetics ; physiopathology ; Humans ; Logistic Models ; Medicine, Chinese Traditional ; Norepinephrine Plasma Membrane Transport Proteins ; genetics ; Promoter Regions, Genetic ; Qi

AIMTo investigate the level of immune response and the immune mechanism of the single-dose hepatitis B surface antigen (HBsAg)-poly (d, l)-lactide-co-glicolide acid (PLGA) microspheres in BALB/c mice.

METHODSThree kind of HBsAg-PLGA microspheres, HBsAg-PLGA50/50-COOH microspheres, HBsAg-PLGA75/25 microspheres and HBsAg-PLGA50/50 microspheres, were prepared by double emulsion microencapsulation technique used three kinds of PLGA with different L/G ratio. The single-dose of HBsAg-PLGA microspheres was subcutaneously injected into BALB/c mice at the dose of 7.5 microg HBsAg per mouse. The conventional aluminum-adjuvant vaccine was subcutaneously injected at 0, 1 and 2 month as positive control. In certain time interval, the induced immune level of total antibody was detected by enzyme linked immunosorbent assay (ELISA). For subclass of IgG antibody and cytokines studies, the dose of HBsAg was 2.5 microg per mouse.

RESULTSThe HBsAg-PLGA microspheres could successfully induce a humoral immune response in BALB/c mice. Compared with the conventional aluminum-adjuvant vaccine, the antibody response of the HBsAg-PLGA50/50-COOH microspheres was significantly lower than the group received three injections of aluminum-adjuvant vaccine (P < 0.01) except for a higher priming response during the early 6 weeks. The results were ascribed to the relatively rapid degradation charactics of PLGA50/50-COOH polymer. The immune response for the HBsAg-PLGA50/50 microspheres and HBsAg-PLGA75/25 microspheres were comparable to the group administered with aluminum-adjuvant vaccine (P > 0.05) which was due to the sustained degradation of PLGA50/50 and PLGA75/25 polymer.

CONCLUSIONThe HBsAg-PLGA microsphere is a promising candidate for the controlled delivery of a vaccine which does not require multiple injections.

Animals ; Delayed-Action Preparations ; Dose-Response Relationship, Immunologic ; Drug Carriers ; Female ; Hepatitis B Antibodies ; blood ; Hepatitis B Surface Antigens ; administration & dosage ; chemistry ; immunology ; Hepatitis B Vaccines ; administration & dosage ; immunology ; Immunization ; Immunoglobulin G ; blood ; Interferon-gamma ; metabolism ; Interleukin-2 ; metabolism ; Interleukin-5 ; metabolism ; Lactic Acid ; Mice ; Mice, Inbred BALB C ; Microspheres ; Polyglycolic Acid ; Polymers ; Random Allocation ; Rats ; Rats, Wistar

OBJECTIVETo explore the effects of connective tissue growth factor (CTGF) on the pathogenesis of human keloid.

METHODSCTGF antisense oligonucleotides (ASODN) conjugated with isothiocyanate fluorescence was encapsulated by liposome, and then added into the human keloid fibroblast (HKF) culturing media. The intracellular distribution of CTGF ASODN was observed with fluorescence microscopy in the fixed HKF. The proliferation of HKF was measured by MIT test. The apoptosis of HKF was measured with a flow cytometer. The collagen synthesis of HKF was measured by using H3-proline incorporation method.

RESULTSThe CTGF ASODN inhibited the proliferation and collagen synthesis of the HKF, compared with the control, but it increased the apoptosis after the transfection (P < 0.01).

CONCLUSIONCTGF ASODN may has anti-fibrotic effects on human keloid in vitro, and the CTGF may play an important role in promoting the fibrosis of human keloid.

Apoptosis ; Cell Differentiation ; Cells, Cultured ; Connective Tissue Growth Factor ; genetics ; Fibroblasts ; cytology ; Humans ; Keloid ; metabolism ; pathology ; Oligonucleotides, Antisense ; genetics ; Transfection

OBJECTIVETo explore the effect of connective tissue growth factor on the pathogenesis of human hypertrophic scar.

METHODSNormal skin and hypertrophic scar fibroblasts were cultured in vitro. The collagen synthesis of fibroblasts were measured by H3-proline incorporation method. The expression of connective tissue growth factor protein and mRNA of fibroblasts were detected with immunocytochemistry staining and reverse transcription polymerase chain reaction methods.

RESULTSCompared with normal skin fibroblast, the collagen synthesis and the expression of connective tissue growth factor protein and mRNA in the hypertrophic scar fibroblast was higher (P < 0.01).

CONCLUSIONConnective tissue growth factor may play an important role in promoting the fibrotic process of hypertrophic scar.

Cells, Cultured ; Cicatrix, Hypertrophic ; genetics ; metabolism ; pathology ; Collagen ; biosynthesis ; Connective Tissue Growth Factor ; Fibroblasts ; metabolism ; pathology ; Gene Expression ; Humans ; Immediate-Early Proteins ; genetics ; Immunohistochemistry ; Intercellular Signaling Peptides and Proteins ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

BACKGROUNDFanconi anemia is a severe congenital disorder associated with mutations in a cluster of genes responsible for DNA repair. Arriving at an accurate and timely diagnosis can be difficult in cases of Fanconi anemia with atypical clinical features. It is very important to increase the rate of accurate diagnosis for such cases in a clinical setting. The purpose of this study is to explore the clinical diagnosis of Fanconi anemia in children with atypical clinical features.

METHODSSix cases of Fanconi anemia with atypical clinical features were enrolled in the study, and their clinical features were recorded, their FANCA gene transcription was assessed by RT-PCR, and FANCA mutations and the ubiquitination of FANCD2 protein were analyzed using DNA sequencing and western blotting respectively.

RESULTSAll six cases showed atypical clinical features including no apparent deformities, lack of response to immune therapy, and progressively increasing bone marrow failure. They also have significantly increased fetal hemoglobin, negative mitomycin-induced fracture test results, and carry a FANCA gene missense mutation. Single protein ubiquitination of FANCD2 was not observed in those patients.

CONCLUSIONThe combination of clinical features, FANCA pathogenic gene mutation genotype and the absence of FANCD2 protein ubiquitination are helpful in the accurate and timely diagnosis of Fanconi anemia in children.

Child ; Child, Preschool ; Fanconi Anemia ; diagnosis ; genetics ; metabolism ; Fanconi Anemia Complementation Group D2 Protein ; genetics ; metabolism ; Female ; Humans ; Male ; Mutation ; Ubiquitination

OBJECTIVETo study the role of connective tissue growth factor (CTGF) in the pathogenesis of human keloid.

METHODSHuman keloid fibroblasts (HKF) were isolated from human keloid and cultured in vitro. The cells were then divided into 3 groups according to different processing, i.e. ASODN treatment (AT), in which phosphorothioate CTGF antisense oligonucleotides (ASODN) labeled by fluorescent isothiocyananate were transfected into the HKFs by liposome; liposome control (LC, with liposome only); control groups (without liposome or ASODN). The distribution of CTGF ASODN in all groups of cells was observed under fluorescent microscope. The CTGF mRNA index (RI) of HKF was assessed by reverse transcription polymerase chain reaction method (RT-PCR). The collagen synthesis of HKF was assessed by (3)H-proline incorporation method.

RESULTSA large amount of fluorescence could be observed in the cytoplasm of HKFs in AT 12 hours after transfection, but not in LC and C groups. The CTGF mRNA index of HKF in AT group 48 hours after transfection was significantly lower than that in LC and C groups (0.12 +/- 0.62 vs 0.51 +/- 0.18 vs 0.54 +/- 0.35, P < 0.01). The (3)H-proline incorporation rate in AT group (108.96 +/- 79.05) was lower than that in LC and C groups (P < 0.01).

CONCLUSIONThe expression of CTGF gene and collagen synthesis of the cultured HKF could be inhibited by CTGF ASODN, implying that CTGF played a role in the development of excessive fibrosis of human keloid.

Collagen ; biosynthesis ; Connective Tissue Growth Factor ; Fibroblasts ; metabolism ; Humans ; Immediate-Early Proteins ; antagonists & inhibitors ; genetics ; physiology ; Intercellular Signaling Peptides and Proteins ; genetics ; physiology ; Keloid ; etiology ; metabolism ; Oligonucleotides, Antisense ; pharmacology ; RNA, Messenger ; analysis ; Transfection

AIMObserve the effects of er-long-zuo-ci-wan (EIZCW, a compound of Chinese Traditional Medicine) on the spontaneous discharge of external cortex of inferior colliculus (ICx) and secondary auditory cortex (AII) of chronic tinnitus model rats induced by salicylate acid, to explore the neural mechanisms underlying ELZCW preventing tinnitus.

METHODS30 adult SD rats were involved and divided into three groups, normal control group, chronic tinnitus model group and ELZCW prevention group. Extracellular recording techniques and stereotaxic method were used. The spontaneous spikes were recorded and analyzed from ICx and all in different group rats. The average rate of spontaneous discharge and the interspike interval histogram of spontaneous activities were used as indexes.

RESULTS(1) Compared with normal control group, the average rate of spontaneous discharge recorded from the ICx in the chronic tinnitus model group increased significantly (4.57 +/- 0.54 Hz vs. 3.14 +/- 0.40 Hz, P < 0.05). Furthermore analysis showed that the discharge rate of short spike interval from the ICx in the chronic tinnitus model group increased than that of the normal group (0-40 ms: 58% vs. 40%; 0-4 ms: 9% vs. 5%). And there was an increasing tendency of the average rate of spontaneous discharge recorded from the AII in the chronic tinnitus model group compared with that in the normal group. (2) Compared with the chronic tinnitus model group, the average rate of spontaneous discharge recorded from the ICx and AII in the ELZCW prevention group significantly decreased than that in the chronic tinnitus model group (ICx: 2.41 +/- 0.21 Hz vs. 4.57 +/- 0.54 Hz, P < 0.01. AII: 2.24 +/- 0.24 Hz vs. 4.57 +/- 0.54 Hz , P < 0.01). And the discharge rate of short spike interval from the ICx and AII in the chronic tinnitus model group decreased than that in the normal control group (ICx: 0-40 ms 50% vs. 58%, 0-4 ms 4% vs. 9%. All: 0-22 ms: 24% vs. 31%, 0-8 ms 19% vs. 16%).

CONCLUSIONIf the increasing of the spontaneous activities of ICx and AII in chronic tinnitus rats means tinnitus, the use of ELZCW could decrease this kind of changes.

Animals ; Auditory Cortex ; physiopathology ; Auditory Pathways ; drug effects ; physiopathology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Male ; Phytotherapy ; Rats ; Rats, Sprague-Dawley ; Salicylic Acid ; Tinnitus ; chemically induced ; drug therapy ; physiopathology