Objective: To establish capillary zone electrophoresis method for determination of sildenafil citrate (Viagra) content in its troche. Methods： Using tetrandrine as internal standard(IS), the electrophoretic separation was achieved with 25 mmol/L borate (pH=7.89) running buffer. And a voltage of 14 kV was applied to the 40 cm×75 μm(i.d) capillary. The analytes were introduced into capillary by siphon (1 s) and determined with on-column monitoring at 214 nm. Results：The determination could be completed within 4 min and the minimum concentration of detection was 5 μg/ml.The analytical results of sildenafil citrate samples demonstrated a good linear relationship within the range of 24-480 μg/ml.The relative standard deviations (RSD) of intra-day was less than 1.58% and that of inter-day was less than 2.46%.The present recoveries were between 95%-105％. Conclusion：The CZE method is accurate, simple, rapid and reliable, so it can be applied to the determination of sildenafil citrate content.

Because the ureter arises from the mesonephric or Wolffian duct (WD), the WD opening should migrate inferiorly along the urogenital sinus or future urethra. However, this process of descent has not been evaluated morphometrically in previous studies and we know little about intermediate morphologies for the descent. In the present work, serial sagittal sections of 15 specimens at gestational age 6–12 weeks and serial horizontal sections of 20 specimens at 6–10 weeks were analyzed. Monitoring of horizontal sections showed that, until 9 weeks, a heart-, lozenge- or oval-shape of the initial urogenital sinus remained in the bladder and urethra. Thus, the future bladder and urethra could not be distinguished by the transverse section or plane. The maximum width of the urogenital sinus or bladder at 6–10 weeks was 0.8 mm, although its supero-inferior length reached 5 mm at 10 weeks. During earlier stages, however, the medial shift of the WD was rather evident. Depending on the extent of upward growth of the bladder smooth muscle, the descent of the vas deferens became evident at 10–12 weeks. Development of the urethral rhabdosphincter likely resulted in the differentiation of urogenital sinus into the urethra and bladder before formation of the bladder neck with 3-layered smooth muscles. Development of the prostate followed these morphological changes, later accelerating the further descent of the WD opening. Because of their close topographical relationships, slight anomalies or accidents of the umbilical cord at 10–12 weeks may have a significant effect on normal anatomy.
Embryonic Structures* ; Fetus* ; Gestational Age ; Human Development ; Humans* ; Muscle, Smooth ; Neck ; Prostate ; Umbilical Cord ; Ureter ; Urethra ; Urinary Bladder ; Vas Deferens* ; Wolffian Ducts*

The raphe of the human penis and scrotum is considered to develop secondarily after disappearance of the initial midline seam by fusion of the bilateral genital folds. However, the fetal development was still obscure. We examined histological sections of 30 fetuses (17 males and 13 females) at 10-15 weeks. In male fetuses, the scrotum was not yet clearly identified because of no descent of testis. The perineal raphe was thin and wavy at 10 weeks, and it was continuous with and took a direction same as the inferior wall of the closed penile urethra after physiological hypospadias. Depending on growth of the bulbospongiosus muscle and corpus spongiosus penis, the midline intermuscular septum obtained a connection to the subcutaneous wavy raphe and made the latter thick and straight at 12-15 weeks. Notably, the perineal raphe extended posteriorly to attach to the external anal sphincter. In female fetuses, an epithelial fusion occurred along a short distance at the posterior end of the vestibule. However, in front of the external anal sphincter, a large midline mesenchymal tissue from the urorectal septum did not contain a raphe-like structure. Moreover, since the bilateral bulbospongiosus muscles were separated widely by the vestibule, they did not provide a midline septum. Fetal development of the perineal raphe was accelerated by reinforcement from the muscular septum. In contrast, without such a muscular support, the female raphe could not maintain its growth even if the seed appeared at the posterior end of the vestibule.
Anal Canal* ; Female ; Fetal Development ; Fetus* ; Humans* ; Hypospadias ; Male ; Muscles ; Penis ; Scrotum ; Testis ; Urethra

OBJECTIVEFungal keratitis (FK) is a vision-threatening infection, whose treatment requires more effective and safer anti-fungal agent exploitation urgently. With this aim, we focused on the effect of an extracellular polysaccharide on fungal adhesion to human corneal epithelial cells.

METHODSWe performed the cytotoxicity assays of the extracellular polysaccharide EPS-II from an antarctic bacterium Pseudoaltermonas and evaluated its inhibitory effect on Candida albicans cells' adherence to human corneal epithelial cells (HCECs).

RESULTSEPS-II, which displayed minor cytotoxicity but also promoted proliferation of HCECs, could inhibit the adherence of yeast cells to HCECs in a dose-dependent manner. EPS-II could also suppress the subsequent PI3K/AKT signaling pathway, and thereby decrease the expression of early inflammatory cytokines.

CONCLUSIONSExtracellular polysaccharide EPS-II was suggested as a new natural agent for attenuating FK.

Blotting, Western ; Candida ; drug effects ; physiology ; Cell Adhesion ; drug effects ; Cornea ; microbiology ; Humans ; Phosphorylation ; Polysaccharides, Bacterial ; pharmacology ; Proto-Oncogene Proteins c-akt ; metabolism ; Pseudoalteromonas ; metabolism

The therapeutic effect of sustained intravitreal injectable voriconazole microspheres (VCZ-MS) on an experimental endophthalmitis of Aspergillus fumigatus was investigated. VCZ-MS was prepared successfully and its physico-chemical property was also evaluated. Right eyes of albino rabbits were infected with an intravitreal injection of 1 000 CFU x mL(-1) of susceptible Aspergillus fumigatus. All fungal endophthalmitis models were randomly divided into five groups 48 hours later: Group A is control group with no treatment; in group B, vitrectomy was performed combined with intravitreal 3 times injections of 100 microg x 0.1 mL(-1) voriconazole every other day. In group C, D and E, vitrectomy was performed combined with intravitreal injection of 0.5 mg, 1.0 mg and 1.5 mg VCZ-MS respectively. The treatment effect was assessed by slit lamp and indirect ophthalmoscope funduscopy examination, using clinical grading system of inflammation in the anterior chamber and the vitreous opacity. The optical microscopy revealed that microspheres obtained from the experiment design were opaque, discrete and spherical particles with smooth surfaces. The drug content and encapsulation efficiency of microspheres were 29.94% and 73.5%, respectively. Endophthalmitis occurred in all eyes of group A, and rapidly developed to panophthalmitis. The inflammation grade of group B, C, D or E was lower than that of group A (P < 0.05). The grade of vitreous opacity in group C, D, E is lower than group B (P < 0.05). Two eyes in group C developed to panophthalmitis. But in group D and E, all eyes whose inflammation was controlled had no recurrence with vitreous clear. Histopathological examination showed normal structures in the cured eyes, while most uncured eyes were atrophic and with eyeball destroyed. So, it can be safely concluded that the curative effect of intravitreal VCZ-MS is significantly better than that of routine intraocular injection of voriconazole. The optimal dose is the one containing 1.0 mg voriconazole.
Animals ; Antifungal Agents ; administration & dosage ; therapeutic use ; Aspergillosis ; drug therapy ; pathology ; Aspergillus fumigatus ; Delayed-Action Preparations ; Endophthalmitis ; drug therapy ; microbiology ; pathology ; Eye ; microbiology ; pathology ; Female ; Intravitreal Injections ; Lactic Acid ; Male ; Microspheres ; Polyglycolic Acid ; Pyrimidines ; administration & dosage ; therapeutic use ; Rabbits ; Random Allocation ; Triazoles ; administration & dosage ; therapeutic use ; Voriconazole

To investigate the inhibitory effect of Pluronic on P-glycoprotein (P-gp) drug efflux pump, Caco-2 cells and animal models were established to study the influence of Pluronic on celiprolol transport across Caco-2 cell monolayer and intestinal mucous membrane with verapamil set as a positive control. Drug concentration was measured by HPLC and the apparent permeability coefficient (P(app)), absorption rate constant (k(a)) and the effective permeability coefficient (P(eff)) were calculated. P(app) of basolateral to apical side and apical to basolateral side was (2.10 +/- 0.13) x 10(-6) and (0.333 +/- 0.018) x 10(-6) cm x s(-1), respectively. Transports of celiprolol across Caco-2 cell monolayer were influenced by both verapamil and Pluronic. The absorption constants (k(a)) of celiprolol at duodenum, jejunum, ileum, and colon were (0.09 +/- 0.03), (0.14 +/- 0.04), (0.11 +/- 0.03) and (0.05 +/- 0.02) h(-1), k(a) of celiprolol in verapamil group were (0.14 +/- 0.03), (0.24 +/- 0.02), (0.25 +/- 0.03) and (0.23 +/- 0.02) h(-1), and k(a) of celiprolol in Pluronic group were (0.13 +/- 0.02), (0.22 +/- 0.02), (0.22 +/- 0.03) and (0.20 +/- 0.03) h(-1), respectively. Pluronic showed significant effect on inhibiting P-gp of Caco-2 cell and intestinal mucosa in rats.
ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Animals ; Biological Transport ; drug effects ; Caco-2 Cells ; Celiprolol ; pharmacokinetics ; Excipients ; Humans ; Intestinal Absorption ; drug effects ; Intestinal Mucosa ; metabolism ; Jejunum ; metabolism ; Male ; Permeability ; Poloxamer ; administration & dosage ; pharmacology ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the role of late course accelerated fractions.ted irradiation(LCAF) combined with concurrent chemotherapy in the management of esophageal carcinoma.Methods From March 1998 to July 2000,111 eligible patients were randomized into LCAF alone group(LCAF,57 patients)or LCAF plus concurrent chemotherapy group(LACF-CT,54 patients).The radiotherapy regimen was identical in the two groups,consisting of conventional fractionation in the first 2/3 course and accelerated fractionation in the second 1/ 3 course to a total dose of 68.4 Gy/41 Fx/44 d.Chemotherapy regimen consisted of four eycles of cisplatin 25 mg/ (m~2?d)plus fluorouracil 600 mg/(m~2?d)on day 1 to 3 every 4 weeks and was delivered on the first day of radiotherapy.Results The median follow-up time was 67.1 months(range 47.6-76.4 months).The 1-,3-,5- year survival rate was 67%,44% ,40% and 77%,39% 28% in LACF-CF and LEAF group,respectively(P =0.310).Grade 3+4 acute side-effact was 42% and 25% in LCAF-CT and LCAF group,respectively(P＜0. 05),with 3 treatment-related deaths in the LCAF-CT group.Conclusions Late course accelerated fractionated irradiation combined with concurrent chemotherapy has a trend towards improving the survival,at the cost of increasing acute side-effect.Its role needs further confirmation by larger sample studied in randomization.

Objective Toinvestigatethecontentofintrinsicformaldehydeinsea-food,soastoprovideascientificbasis forestablishingthecriteriaforformaldehyderesidueinsea-foods.Methods Atotalof266samplesofvarioussea-foods were collected from 3 representative coastal regions in Zhejiang Province.The content of intrinsic formaldehyde was examined at pre-determined gradients based on different storage temperature and time.A total of 825 test results were collected.All samples were carefully protected against contamination of exogenous formaldehyde in the process from collectiontoexamination.Results Theconcentrationofformaldehyderangedbetween10.40and223.80,0.08and3.50, 0.08 and 1 4.20,0.08 and 2.89,0.25 and 1 1.02,0.1 2 and 2.61 ,0.08 and 2.37 mg/kg for Bombay duck,yellow croaker,squid,hairtail,inkfish,octopus,and shrimp respectively.The intrinsic formaldehyde content varied between different sea -foods,and also between different individuals of a same species.Meanwhile,storage time significantly influencedintrinsicformaldehydecontentofsea-foods.Conclusion Sea-foodscontainedcertainamountofintrinsic formaldehyde.The national standard which requires zero content of formaldehyde in marine products needs further discussion.

Umbilical metastases from intraperitoneal malignancies are universally referred to Sister Mary Joseph's nodule (SMJN). The most frequent primary tumor sites include the stomach and ovaries. SMJN caused by colon cancer is uncommon. Likewise, carcinoma of the right side colon metastasizing to inguinal lymph nodes is considered almost impossible. To the best of our knowledge, there is no report of right side colon cancer synchronously involving both the umbilicus and inguinal lymph nodes in the literature. We present a case of right side colon cancer (RSCC) metastasizing to the umbilicus and inguinal lymph nodes, which was confirmed by routine pathological evaluation and immuohistochemistry.
Adenocarcinoma ; pathology ; secondary ; surgery ; Adult ; Carcinoembryonic Antigen ; blood ; metabolism ; Colectomy ; methods ; Colonic Neoplasms ; pathology ; surgery ; Groin ; Humans ; Keratin-20 ; metabolism ; Lymph Node Excision ; Lymph Nodes ; pathology ; surgery ; Lymphatic Metastasis ; Male ; Sister Mary Joseph's Nodule ; pathology ; secondary ; surgery

OBJECTIVETo reproduce a model of heat injured KC in vitro and explore its apoptosis rate of KC due to heat injury at different temperature.

METHODSHuman KCs were cultured in vitro, and they were incubated at 37, 41, 43, 45, 48, and 51 degrees C respectively for 10 mins in water bath. Trypan blue staining and Hoechst 33258 fluorescence staining were used respectively to determine necrosis and apoptosis of KC. Rates of apoptosis and necrosis of KC were analyzed quantitatively by flow cytometer. The proliferation activity of KC after heat injury was detected by MTT test.

RESULTSThe results of trypan blue staining, Hoechst 33258 fluorescence staining, and flow cytometer demonstrated that number of apoptotic and necrotic KC increased gradually along with a rise of water bath temperature. The rates of apoptosis and necrosis of KC were respectively (12.3 +/- 3.2)% and (14.1 +/- 1.6)% at 45 degrees C, (27.7 +/- 5.1)% and (58.0 +/- 4.2)% at 48 degrees C. Rate of KC necrosis reached up to (83.0 +/- 5.3)% at 51 degrees C. Inhibition of KC growth reached a stationary phase when the injurious temperature reached 45 degrees C as observed with MTT test.

CONCLUSIONSHeat injury can induce apoptosis and growth inhibition of KC in vitro. Incubating KC at 45 degrees C for 10 mins is a good condition to reproduce a model of heat injured KC in vitro. This model may be used to study the biological character and apoptosis of KC after burn injury.

Apoptosis ; Burns ; Cell Proliferation ; Cell Survival ; Cells, Cultured ; Flow Cytometry ; Hot Temperature ; Humans ; Keratinocytes ; cytology