Sperm ultrastructural abnormalities are often associated with sperm motility, the integrity of genetic material, and the fertilization potential. The investigation of sperm ultrastructural abnormalities is based on the evolution of microscopy techniques. In his paper, we review the improvement of the microscopy techniques and the ultrastructure of several specific morphological defects and he apoptotic spermatogenic cells in order to expound the significance of sperm ultrastructural observation in clinical practice. We deem it necessary to analyze the sperm ultrastructure before exploring the pathology and adopting assisted reproductive technology for some special patients with teratozoospermia.
Humans ; Male ; Microscopy ; Spermatozoa ; abnormalities ; ultrastructure

AIM:To investigate the cause of blindness, except those caused by cataract, in Haimen city.METHODS:According to the WHO`s criteria of blindness, the blindness level was decided through ophthalmic tests by associate chief or chief ophthalmologists who were trained especially for disability evaluation.The analysis of the the leading cause were taken too.RESULTS:Totally 3 266 persons were blindness, in which 2 118 were first level blindness, 1 148 persons were second lever blindness, and 1 308 persons were male, 1958 were female.The leading cause of blindness were retina and uveitis diseases (31.58%), genetic diseases(23.47%), cornea disease(14.49%).CONCLUSION:The leading cause of blindness are retina and uveitis diseases, genetic diseases, cornea diseases in Haimen city of Jiangsu province.Early prevention and treatment should be strengthened to reduce the occurrence of blindness.

Objective: To analyze the long-term prognosis of re-operation in patients with functional tricuspid regurgitation (FTR) after left sided valve replacement (LSVR) and hence evaluate the optimal timing of mentioned re-operation. Methods: A total of 59 FTR patients who had re-operation after their prior LSVR in our hospital from 1999-01 to 2013-01 were analyzed. The clinical information and post-operative follow-up results were recorded in all patients. Results: There were 5/59 (8.5%) patients died in peri-operative period and the overall post-operative mortality was 11.9% (7/59). The follow-up data of 54 survivors were available for the mean time of 51.1 (21-188) months. There were 19/54 (35.2%) patients suffered from MACE and 30 (55.6%) were beneifted by improved cardiac function. Uni-variable analysis indicated that pre-operative NYHA class IV (P=0.008), pre-operative right ventricular (RV) dysfunction (P=0.037), concomitant left-sided redo-operation (P=0.017) and TVR operation (P=0.002) were associated with all cause mortality of tricuspid re-operation. Multi-variable Cox regression analysis showed that pre-operative RV dysfunction was the only independent risk factor of long term MACE-free accumulating survival rate (HR=3.0, 95% CI 1.11-8.2,P=0.031); while TVR operation (HR=12.8, 95% CI 1.53-107.02,P=0.019) and pre-operative NYHA class IV (HR=5.3, 95% CI 1.20-24.51,P=0.032) were the independent risk factors for long-term mortality in patients after tricuspid re-operation. Conclusion: Patients with compensatory RV function showed better long term prognosis after secondary tricuspid operation. Aggressive re-operation before the occurrence of right ventricular dysfunction could be beneficial for relevant patients.

Objective To investigate the effects of different hepatic perfusion procedures for small-for-size liver transplantation in rats. Methods A total of 156 rats were randomly divided into two groups: portal vein perfusion group (groupⅠ, n=78) and abdominal aorta perfusion group (groupⅡ, n=78). After harvesting graft, the left lobe of the liver and the middle lobe were resected and the remaining approximately 30%volumes of the liver were transplanted in groupⅠand groupⅡ. The body weights of donor and acceptor, the weight of graft, the time of operating in donor, the cold ischemia time, anhepatic phase, the blocking time of inferior hepatic vena cava and the time of operating in receptor were recorded in two groups. The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), pathological HE staining and 7-day survival rate in 6 h, 1 d, 3 d and 7 d after operation were compared between two groups. Results The serum levels of ALT and AST were decreased gradually in two groups, but the levels decreased slowly in groupⅠ. The serum levels of ALT and AST were significantly higher in groupⅠthan those of groupⅡ(P<0.05). HE staining showed greater damage of mi-crostructure of liver tissue at early stage in group Ⅰthan that in groupⅡ. The 7-day survival rate was lower in group Ⅰthan that of groupⅡ(χ2=4.050,P=0.044). Conclusion There is a higher survival rate and mild liver damage in small-for-size liver transplantation in rats using perfusion by abdominal aorta.

Objective To investigate the molecular mechanisms of long non-coding RNA ZEB1-antisense RNA1 (LncRNA ZEB1-AS1) in regulating lymphoma cell proliferation, invasion, and migration by targeting microRNA (miR)-224. Methods The lymphoma cell line Raji was cultured in vitro and transfected with si-NC, si-ZEB1-AS1, miR-NC, miR-224 mimics, si-ZEB1-AS1+anti-miR-NC, and si-ZEB1-AS1+anti-miR-224, respectively. Real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) was used to assess the expression levels of LncRNA ZEB1-AS1 and miR-224 in lymphoma cells. Cell viability was detected using the Cell Counting Kit-8 (CCK-8). Cell migration and invasion abilities were evaluated by Transwell experiments. Dual luciferase reporter experiments were conducted to confirm the regulatory effect of LncRNA ZEB1-AS1 on miR-224. Western blot was performed to detect the expression levels of cyclin D1 (CyclinD1), matrix metalloproteinase (MMP)-2, and MMP-9 proteins. Results Compared with the control group, the lymphoma group showed increased LncRNA ZEB1-AS1 levels and decreased miR-224 expression levels (P < 0.05). Compared with the si-NC group, the si-ZEB1-AS1 group exhibited decreased expression levels of LncRNA ZEB1-AS1, optical density (OD) values, the number of migrated and invaded cells, and expression levels of CyclinD1, MMP-2, and MMP-9 protein (P < 0.05). Compared with the miR-NC group, the miR-224 group showed increased miR-224 expression levels but decreased OD values, the number of migrated and invaded cells, and expression levels of CyclinD1, MMP-2, and MMP-9 proteins (P < 0.05). Upregulation of miR-224 levels reduced the luciferase activity of WT-ZEB1-AS1 but had no effect on MUT-ZEB1-AS1 luciferase activity. Compared with the pcDNA-NC group, the pcDNA-ZEB1-AS1 group showed decreased miR-224 expression levels (P < 0.05); compared with the si-NC group, the si-ZEB1-AS1 group showed increased miR-224 expression levels (P < 0.05). Compared with the si-ZEB1-AS1+anti-miR-NC group, the si-ZEB1-AS1+anti-miR-224 group showed increased OD values, the number of migrated and invaded cells, and expression levels of CyclinD1, MMP-2, and MMP-9 proteins(P < 0.05). Conclusion Inhibition of ZEB1-AS1 to targetedly upregulate of miR-224 expression can effectively reduce lymphoma cell proliferation, invasion, and migration abilities.

Background Femtosecond laser in situ keratomileusis (LASIK) inevitably injury keratocytes and corneal nerve fibers.The research report about postoperative morphological changes of corneal nerve regeneration and keratocytes in femtosecond LASIK is still rare.Objective The aim of this study was to observe the kinetic changes of keratocytes and corneal nerve in corneal flap after femtosecond LASIK.Method Femtosecond laser manufacture corneal flap of LASIK surgery was performed on 60 eyes of 30 patients with refractive error using both femtosecond laser system and excimer laser treatment system.The repair of corneal wound was examined by slit lamp microscope,and the morphology of keratocytes and corneal nerve were observed with confocal microscope 1 week,1month,3 months after surgery,respectively.Results No haze or flap folds were found under the slit lamp microscope from 1 week through 3 months after operation.One week after surgery,the corneal stromal cells at the interface of the corneal flap appeared to be a mild activation status in 42 eyes (70％),but the activated cells gradually reduced with lapse of time.Three months after surgery,mild activation state still was found in 7 eyes (12％).One week after surgery,independent,short (＜50 μm),curved subbasal nerve fibers were exhibited in 7 eyes (12％),and curved filamentous nerve fibers were discovered in 48 eyes (80％) one month after surgery.The nerve fiber length of subbasal nerve was ＞200 μm in 27 eyes (45％) and classes beaded structure appeared 3 months after operation but were still different with preoperative subbasal nerve fibers.One week after the operation,filaments or discontinuous nerve fibers could been seen in 46 eyes (77％) at theinterface,and long nerve fibers or filamentous nerves were visible around the terminal or periphery of nerve fibers in 49 eyes (82％) one month after surgery,and long nerve fibers or filaments of nerve fibers were visible in 57 eyes (95％) 3 months after the surgery.Conclusions Femtosecond LASIK cause wound reaction at cellular level.Corneal nerve fibers recover with the extension of time,but there are still some morphological differences 3 months after surgery from preoperation.

OBJECTIVETo observe the curative efficacy of rhubarb, montmorillonite powder combined with blood purification on treatment for patients with severe acute organophosphorus pesticide poisoning (AOPP).

METHODS39 patients with AOPP were divided into treatment group (received the combined treatment of rhubarb, montmorillonite powder and blood purification on the basis of routine therapy, n = 21) and control group (only received the routine treatment because of financial difficulties or the will of family members, n = 18). The differences of clinical manifestations, curative effects and prognosis between two groups were compared.

RESULTSThe time of consciousness recovery, the duration of mechanical ventilation and the length of stay in hospital in treatment group were (6.5 ± 1.3), (7.9 ± 2.0) and (13.1 ± 3.2) days, which were significantly shorter than those [(8.4 ± 2.4), (10.7 ± 2.9) and (16.5 ± 3.7) days] of control group (P < 0.05). In 5, 6 and 7 day after treatment,the cholinesterase (ChE) activities of treatment group were significantly higher than those of control group (P < 0.05). The total amount and using time of atropine and pyraloxime methylchloride in treatment group were significantly smaller and shorter than those in control group (P < 0.05). The death rate of treatment group was [19.0% (4/21)], which were significantly lower than that of control group [19.0% (4/21)] (P < 0.05).

CONCLUSIONThe combined treatment of rhubarb, montmorillonite powder and blood purification of the AOPP patients has a better curative effect.

Adolescent ; Adult ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hemofiltration ; methods ; Humans ; Insecticides ; poisoning ; Male ; Middle Aged ; Organophosphate Poisoning ; therapy ; Pesticides ; poisoning ; Phytotherapy ; Prognosis ; Rheum ; Young Adult

This study is to investigate the cytoprotective role of NaNO2 preconditioning against ethanol induced damage in human hepatoma SMMC-7721 cells. The cells were preconditioned with NaNO2 (0.25 mmol x L(-1)) for 24 hours or 4 weeks, and then exposed to ethanol (200 mmol x L(-1)) for additional 12 h and untreated cells served as control. Both temporal and chronic NaNO2 preconditioning could prevent ethanol elicited cytotoxicity as evidenced by thiazolyl blue (MTT). NaNO2 preconditioning also could inhibit ethanol-induced apoptosis, which was confirmed by FITC-Annexin V/PI flow cytometer and Hoechst 33258 and PI staining. Further, simultaneous NaNO2 preconditioning treatment along with ethanol showed protection against ethanol mediated cellular damage as indicated by significantly decreased levels of malondialdehyde (MDA) and elevated activities of superoxide dismutase (SOD) and catalase (CAT). Western blotting analysis revealed that in ethanol treated cells preconditioned with NaNO2, the HIF-1alpha and Bcl-2 increased obviously, while the expression of pro-apoptotic proteins, including Bax, Caspase-9, Caspase-3 decreased. The results showed that low doses of NaNO2 preconditioning resistant to ethanol-induced human hepatoma SMMC-7721 cells apoptosis, which mechanism may be related to increased expression of HIF-1alpha in the cells.
Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Catalase ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Ethanol ; toxicity ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Liver Neoplasms ; metabolism ; pathology ; Malondialdehyde ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Sodium Nitrite ; administration & dosage ; pharmacology ; Superoxide Dismutase ; metabolism ; bcl-2-Associated X Protein ; metabolism

This study is to observe the effect of N-(3-phenylallylidene)-6-fluoro-1, 8-(2, 1-propoxy)-7-(4-methylpiperazin-1-yl)-quinolin-4(1H)-one-3-carbonyl hyarazine (FQ16) on apoptosis of hepatocarcinoma SMMC-7721 cells in vitro. With different concentrations of FQ16 at different times used to treat SMMC-7721 cells in vitro, the proliferation of the cells and the inhibition effect of FQ16 on the cell proliferation were examined by MTT assay. Cell apoptosis was determined by Hoechst 33258/PI fluorescence staining, TUNEL and agarose gel electrophoresis method. The effect of FQ16 on topoisomerase II activity was measured by agarose gel electrophoresis using Plasmid pBR322 DNA as the substrate. Mitochondrial membrane potential (MMP, delta psi m) was measured by high content screening image system. The reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the expression changes of Bcl-2 mRNA and Bax mRNA. The caspase-9, caspase-8, caspase-3, p53, Bcl-2 and Bax protein expressions were detected by Western blotting analysis. The results showed that the cell proliferation was inhibited by FQ16 at 0.625 - 10 micromol L(-1) in a time-dose dependent manner. Treatment of SMMC-7721 cells with different concentrations of FQ16 for 24 h increased the percentage of the apoptosis cells obviously (P<0.05), the typical ladder DNA in apoptotic cells and a concomitant dissipation of the mitochondrial membrane potential. Compared with control group, FQ16 influenced obviously DNA topoisomerase II activity, stimulated DNA cleavage and inhibited DNA reunion mediated by topoisomerase II. In addition, FQ16 (3 - 7.39 micromol L(-1)) increased mRNA expression of Bax and protein expression of p53, Bax, caspase-9, caspase-3, separately, and induced cytosolic accumulation of activities caspase-9 and caspase-3, whereas the mRNA and protein expression of Bcl-2 decreased with no change of caspase-8. Therefore it can be concluded that the effects of inhibited topoisomerase II and mitochondrial-dependent pathways were involved in FQ16 induction of apoptosis of SMMC-7721 cells.
Antineoplastic Agents ; administration & dosage ; chemical synthesis ; chemistry ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; DNA Topoisomerases, Type II ; metabolism ; Dose-Response Relationship, Drug ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Membrane Potential, Mitochondrial ; drug effects ; Molecular Structure ; Piperazines ; administration & dosage ; chemical synthesis ; chemistry ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; metabolism ; Tumor Suppressor Protein p53 ; metabolism ; bcl-2-Associated X Protein ; metabolism

This study is to find out the induction by sodium nitrite of epithelial-mesenchymal transition (EMT) in human hepatocellular carcinoma cells, SMMC-7721. After treatment of SMMC-7721 with 0.25 - 25 mmol.L-1 sodium nitrite for 48 h, the assays used include enzyme-linked immunosorbent assay (ELISA) for evaluation of TGF-beta1, IL-6 and IL-8 level in the conditioned medium, phase-contrast microscopy for morphology observation, and scratch wound healing as well as transwell migration assays for measurement of migration and metastatic potential. Additionally, the hallmarks of EMT, p-AKT and its downstream signaling molecules were examined by Western blotting. The results showed that TGF-beta1 secreted by SMMC-7721 elevated significantly in a dose-dependent fashion, whereas the increased IL-8 and IL-6 did not show dose-dependent response. The EMT was induced by exposure of SMMC-7721 with 0.25 mmol.L-1 of sodium nitrite, which was characterized by increased level of Vimentin, decreased E-cadherin and elevated activity of migration and metastatic potential. The results suggest that sodium nitrite could induce SMMC-7721 EMT by increased secretion of TGF-beta1 and IL-8.
Cadherins ; metabolism ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cell Movement ; Dose-Response Relationship, Drug ; Epithelial-Mesenchymal Transition ; drug effects ; Humans ; Interleukin-6 ; secretion ; Interleukin-8 ; secretion ; Liver Neoplasms ; metabolism ; pathology ; NF-kappa B ; metabolism ; Neoplasm Invasiveness ; Proto-Oncogene Proteins c-akt ; metabolism ; Sodium Nitrite ; administration & dosage ; pharmacology ; Transforming Growth Factor beta1 ; secretion ; Twist-Related Protein 1 ; metabolism ; Vimentin ; metabolism