Adult ; Compliance ; Humans ; Male ; Occupational Exposure ; prevention & control ; Population Surveillance ; Quality Control ; Young Adult

Objective To establish a fast and accurate technique of identifying the pachytene spermatocytes,round spermatids and elongating and condensing spermatids during STA-PUT velocity sedimentation.Methods Using STA-PUT velocity sedimentation method to isolate the pachytene spermatocytes, round spermatids and elongating and condensing spermatids from mouse testes.To determine the cell populations` distribution,each tube of cell fraction was then partially transfered to the 96 plate well,and each well was added with acridine orange dye.Then each well was analyzed using fluorescence microscopy.Results Three types of spermatogenic cells can be identified quickly and accurately by it`s specific cytoplasm/nucleus character using the acridine orange dye staining under fluorescence detection.Conclusions A rubust method to quickly and accurately determine the pachytene spermatocytes,round spermatids and elongating and condensing spermatids during STA-PUT velocity sedimentation is successfully developed.

Background The treatment timing and method of amblyopia rely on the plasticity of visual system.Synapsin is a family of presynaptic terminal specific protein.Its role in visual developmental plasticity is below understood.Objective To investigate the dynamic expressions of synapsin (T-synapsin),and phosphorylation of synapsin (p-synapsin Ⅰ a/b) in visual cortex of normal mice and further explore the role of synapsin in plasticity of visual system.Methods Forty-two clean neonatal C57BL/6 mice were collected.The mice were sacrificed at postnatal 7,14,21,28,35,42,60 days respectively to obtain the tissue samples of visual cortex.Expression levels of T-synapsin and p-synapsin in the visual cortex following the ageing were quantitatively detected using Western blot assay.Results The expression of synapsin in normal mice showed a dynamic increase with the ageing.The T-synapsin Ⅰ a/b/β-actin value in visual cortex was (21.32 ± 3.27) ％,(56.27 ± 10.18) ％,(77.05 ± 10.05) ％,(83.75±10.52) ％,(94.69±11.46)％,(98.75±5.86) ％ of adults mice (postnatal 60 days,P60) in the mice of postnatal 7,14,21,28,35,42 days,respectively,showing a significant difference among them (F =69.538,P ＜ 0.001).Compared with the adult mice,the T-synapsin Ⅰ a/b/β-actin value in the mice of P7,P14,P21,P28 was significantly lower (all at P＜0.05),but no significant difference was found between P35 and P60,P42 and P60 (P =0.280,0.798).The development trend of different synapsin subtypes,such as T-synapsin Ⅰ a/b,T-synapsin Ⅱ a,T-synapsin Ⅱ b and T-synapsin Ⅲ a,was not quite the same during the ageing.The expression of T-synapsin Ⅱ a and Ⅲ a increasing more slowly with development,and kept increasing until P60.Significant differences were found among various age of mice in T-synapsin Ⅱ a,Ⅱ b,Ⅲa respectively(F =42.492 55.595,39.172,all at P＜0.001).The p-synapsin Ⅰ a/b level in the visual cortex elevated with the ageing of the mice,and that peaked in P21 mice,which was (2.86±0.17) times more than that in adult mice.After that,the expression level of p-synapsin Ⅰ a/b dropped rapidly.A significant difference was found in the p-synapsin Ⅰ a/b expression among different ages of mice (F =22.620,P ＜ 0.001).Conclusions Synapsin level in visual cortex presents a developmental change which correlated with the onset and decline of the critical period.Synapsin is probably involved in the regulation of neural plasticity in visual cortex in critical period.

AIM: To observe the inhibition of c-Met inhibitor on proliferation of lens epithelial cells (LECs).METHODS: Human's LECs were cultured and hepatocyte growth factor (HGF) and K252a were added to second passage of cells supplied with Dulbecco's modified eagle's medium (DMEM). MTT assay was used to examine the proliferation of LECs, and Western-blot was used to detect the expression change of Bcl-2 and Caspase-3.RESULTS: The photodensity (A) of HGF (50nmol/L) + K252a (30nmol/L) was not significantly different from that of DMEM control (P＞0.05). The expression of Bcl-2 and Caspase-3 were not significantly different from that in the control group.CONCLUSION: K252a, the inhibitor of c-Met, can effectively inhibit the proliferation of LECs.

AIM: To study the role of nitric oxide ( NO ) and γ-aminobutyric acid ( GABA) in the formation of amblyopia by establishing 2 different types of amblyopic models.METHODS:A total of 18 aged 3-week kittens were randomly divided into monocular deprivation, strabismus and normal groups. All types of amblyopia were developed in the experimental eyes that were detected by P-VEP 12wk later. The cats were killed and the immunocytochemistry staining method were applied to observe under the light microscope the changes of distribution and positive cells areas of NO and GABA across the amblyopic retinal, compared to that from the normal cats of identical age.
RESULTS: The P-VEP showed that the amplitude of wave P1 was lower (P<0. 05) and the P1 latent time was longer ( P<0. 05 ) in two types of amblyopic cats than those in the normal cats. Compared to the normal cats, the NO and GABA positive cells areas were obviously reduced ( P<0. 05 ) across the retina in the amblyopic cats. But no significant difference was found between two kinds of amblyopic cats.
CONCLUSION:The NO and GABA play an important role in the formation of amblyopia in the level of retinal.

Objective To evaluate the value of contrast-enhanced ultrasound in superficial lymphadenopathy. Methods Ninty-four superficial enlarged lymph nodes were studied by 2-dimensional, color Doppler ultrasound, and contrast-enhanced ultrasound. Then the contrast-enhanced images were analyzed by Philips Q-LAB software. All the results were compared with pathological diagnosis. Results For the 94 superficial lymph nodes examined,44 were benign,33 were metastases and 17 were lymphomas. The sensitivity, specificity,and accuracy of contrast-enhanced images were 84% ,74% and 790//oo respectively. Contrast-enhanced ultrasound examination showed intense homogeneous enhancement in 39 of 44 benign lymph nodes; high or low homogeneous enhancement in 25 of 33 and 7 of 33 in metastases respectively;intense homogeneous, and scarce enhancement in 6 of 17 and 9 of 17 in lymphomas respectively. Time-intensity curves showed that compared with metastasis lymph nodes and lymphomas, benign lymph nodes had higher peak intensity and larger area under the curve (P<0.01). Conclusions The diagnosis accuracy was significant increased when contrast-enhanced ultrasound was test against conventional ultrasound. The character of contrast agent enhancement and Q-LAB time-intensity curves provide valuable diagnosis information for differential diagnosis of benign,metastasis lymph nodes and lymphomas.

Objective To investigate the mechanism of hearing loss after focal cerebral ischemia-reperfusion injury in rats.Methods A total of 60 healthy male adult SD rats were included in this study and randomly divided into 2 groups, ischemia-reperfusion(I/R) group and sham operated control, with 30 rats in each group.The rats in I/R group were operated for suture-occluded method to establish middle cerebral artery occlusion(MCAO)model, with ischemia for 60 mins followed by reperfusion for 24 hrs.The control group was only to be isolated cervical vessels, with no thread embolism inserted.The auditory brainstem response (ABR) was tested before operation and at 24 hrs post-operation respectively.At 24 hrs post-operation, we scored neurological functions, measured the changes of water content in the brain using the dry-wet weight method, and determined the infarct volume through TTC method.We also evaluated the integrity of blood-brain barrier(BBB) by viewing the exudation of Evans blue and observed the apoptosis of neurocyte by TUNEL method to conclude apoptotic index(AI).The expression of MMP-9,Claudin-5,Occludin and CX-43 were detected by Western blot.Results Compared with the sham group,the neurological function scores,the infarct volume and water content of the brain increased, with the elevated thresholds of ABR significantly and AI went up in I/R group.The expression levels of MMP-9 and CX-43 were significantly up-regulated,but the expressions of Claudin-5 and Occludin were obviously down-regulated.All of the differences above had statistical significances.Conclusion The mechanism of hearing loss after focal cerebral ischemia-reperfusion injury in rats is possibly related to MMPs activation,neurocyte apoptosis,BBB breakage and gap junction damage.

This study aimed to investigate the effects of concentration, intestinal section and borneol on the intestinal absorption of salvianolic acids. The experiment not only studied the intestinal absorption properties of three concentrations of rosmarinic acid, salvianolic acid B and salvianolic acid A at duodenum, jejunum and ileum, but also of salvianolic acids compatible with borneol at different concentrations using single-pass intestinal perfusion model in rat with phenol red as the marker. The results showed that salvianolic acids was stable under weak-acid condition and affected by metabolism enzyme; The Peff and Ka significantly different among three concentrations of rosmarinic acid and salvianolic acid B, whose intestinal absorption were saturated in high concentration, suggesting that the transport mechanisms of rosmarinic acid and salvianolic acid B were similar to active transport or facilitated diffusion; However, there was inconspicuousness in the Peff and Ka of salvianolic acid A at different concentrations, whose absorption was not saturated in high concentration, indicating that the transport mechanisms of salvianolic acid A was passive diffusion; The Peff and Ka in the ileum obviously higher than those in the duodenum and jejunum, namely the ileum was the best absorption section; When concentration of borneol increased, the enhancing effect of intestinal absorption of salvianolic acids increased, but significantly decreased when borneol increased to some degree. The enhancing effect of medium borneol concentration was the optimum. This implied that borneol can enhance the intestinal absorption of salvianolic acids, and the capacity of enhancing effect was influenced by the concentration of borneol.
Animals ; Benzofurans ; isolation & purification ; pharmacokinetics ; Bornanes ; administration & dosage ; pharmacokinetics ; pharmacology ; Caffeic Acids ; isolation & purification ; pharmacokinetics ; Cinnamates ; isolation & purification ; pharmacokinetics ; Depsides ; isolation & purification ; pharmacokinetics ; Dose-Response Relationship, Drug ; Duodenum ; metabolism ; Ileum ; metabolism ; Intestinal Absorption ; Jejunum ; metabolism ; Lactates ; isolation & purification ; pharmacokinetics ; Male ; Perfusion ; methods ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry

OBJECTIVETo investigate the correlation between mutation in EGFR tyrosine kinase domain and tumor response as well as prognosis in advanced stage non-small cell lung cancer (NSCLC) treated with iressa.

METHODSFrom May 2002 to Feb. 2005, iressa was orally administered at a dose of 250 mg once daily for 106 advanced stage NSCLC patients until occurrence of disease progression or intolerable toxicity. Cancer tissue was obtained from these patients, and DNA was extracted for analysis of mutation in exon 18 to 24 of EGFR. Exon 18 to 24 of EGFR were amplified by nest PCR, sequenced and analyzed from both sense and antisence directions.

RESULTSPrimary NSCLC tissue specimens consisted of 25 frozen tissue blocks and 81 paraffin-embedded tumor tissue blocks from 106 consecutive NSCLC patients. Mutation was found to be more frequent in the adenocarcinoma than in the squamous cell carcinoma (35.9% vs 14.3%, P =0.033). Mutation was identified in 32 patients (30.2%). Response rate to iressa was 71.9% in the patients with EGFR mutation versus 13.5% in those without mutation (P <0.01). Compared with the patients without EGFR mutation, those with mutation had longer overall survival (median, 13.45 vs. 5.25 months; P<0.01) and median time to progression (median, 8.35 vs. 3.0 months; P <0.01).

CONCLUSIONEGFR mutation may be positively correlated with the response and survival in advanced stage Chinese NSCLC patient treated with iressa.

Adenocarcinoma ; drug therapy ; genetics ; pathology ; Adolescent ; Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; genetics ; pathology ; Carcinoma, Squamous Cell ; drug therapy ; genetics ; pathology ; Exons ; Female ; Follow-Up Studies ; Humans ; Kaplan-Meier Estimate ; Lung Neoplasms ; drug therapy ; genetics ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Point Mutation ; Prognosis ; Quinazolines ; therapeutic use ; Receptor, Epidermal Growth Factor ; genetics ; Sequence Deletion

Cryptococcosis ; etiology ; pathology ; Glycogen Storage Disease Type I ; complications ; pathology ; Humans ; Infant ; Male ; Meningitis, Fungal ; etiology ; pathology