Search

Background: To effectively block the invasion of human immunodeficiency virus (HIV)-1 on mucosal surface, vaginal anti-HIV-1 microbicides should avoid inflammatory responses and disruption of mucosa integrity because these will facilitate transepithelial viral penetration and replication. However, existing models fail to predict and evaluate vaginal mucosal toxicity induced by microbicides, and most importantly, they are unable to identify subtle or subclinical inflammatory reactions. This study was designed to develop a cost-effective in vivo model to evaluate microbicide safety in a preclinical study which can recapitulate the mucosal topical reaction., Methods: A murine model was employed with nonoxynol-9 (N-9) as the topical stimulant within the vagina. Different concentrations of N-9 (1%, 3%, and 4%) were topically applied to the vagina for five consecutive days. A panel of inflammatory cytokines including interleukine-2 (IL-2), IL-4, IL-6, IL-17A, interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), and immuno-regulatory IL-10 were assayed in vaginal lavage. Cytokines were quantified by using cytometric bead array (CBA) and reverse transcript (RT) real-time PCR. Histopathological evaluation of vaginal tissues was conducted on hematoxylin-eosin stained slides and scored with a semi-quantitative system according to the severity of epithelial disruption, leucocyte infiltration, edema, and vascular injection. The association between the cytokines and histopathological scores was assessed by linear regression analysis., Results: All three concentrations of N-9 induced inflammatory cytokine production. The 4% N-9 application resulted in a consistent production of cytokines in a time-dependent manner. The cytokines reached peak expression on day three with the exception of IL-4 which reached its peak on day one. Histopathological examination of 4% N-9 treated cervicovaginal tissues on day three showed intensive damage in four mice (sores: 10 - 13) and moderate damage in one mouse (score: 8), which were significantly associated with both inflammatory cytokines IL-17A and IL-6 and anti-inflammatory cytokines IL-4 and IL-10. Interestingly, IL-17A showed significant positive association with inflammatory cytokine TNF-α (r = 0.739; P < 0.05), anti-inflammatory cytokines IL-10 (r = 0.804; P < 0.01) and IL-4 (r = 0.668; P < 0.05)., Conclusions: Our data demonstrate that a panel of cytokines (IL-17A, IL-6, IL-4 and IL-10) could be used as surrogate biomarkers to predict the histopathological damage. Th17 may play a central role in orchestrating inflammatory cytokine responses. This Th17 based mouse model is cost-effective and suitable to assess the toxicity of candidate microbicides in preclinical studies.

The photochemical decomposition of perfluorodecanoic acid (PFDeA) in water in the presence of persulfate ion (S(2)O(8)(2-)) and sulfur ion (S(2-)) was investigated under vacuum ultraviolet (VUV) light irradiation. PFDeA was decomposed under VUV light irradiation. With the addition of S(2)O(8)(2-) or S(2-), the photo-decomposition and defluorination of PFDeA were enhanced significantly. Sulfate radical anion (SO(4)(*-)) generated from photolysis of S(2)O(8)(2-) initiated PFDeA oxidation. While the S(2-) ion, acting as a *OH scavenger, enhanced the role of reduction pathway induced by aqueous electrons (e(aq)(-)). The shorter-chain perfluorocarboxylic acids (PFCAs), formed in a stepwise manner from longer-chain PFCAs, were identified as products by HPLC/MS., (Copyright 2010 Elsevier B.V. All rights reserved.)

The photochemical decomposition of perfluorooctanoic acid (PFOA) in aqueous periodate (IO(4)(-)) was investigated under two types of low-pressure mercury lamps: one emits at 254nm light (UV light) and the other emits both 254 nm and 185 nm light (VUV light). PFOA decomposed efficiently under VUV light irradiation while it decomposed poorly under UV light irradiation. The addition of IO(4)(-) significantly increased the rate of decomposition and defluorination of PFOA irradiated with UV light whereas it decreased both processes under VUV irradiation. Reactive radical (IO(3)) generated by photolysis of IO(4)(-) initiated the oxidation of PFOA in UV process. Aquated electrons (e(aq)(-)), generated from water homolysis, scavenged IO(4)(-) resulting in decrease of reactive radical species production and PFOA decomposition. The shorter-chain perfluorocarboxylic acids (PFCAs) formed in a stepwise manner from long-chain PFCAs., (2010 Elsevier B.V. All rights reserved.)

Infections with influenza A viruses are still a major threat to humans and several animal species. The occurrence of highly pathogenic avian influenza viruses capable of infecting and killing humans highlights the urgency for a new and efficient strategy for the treatment of diseases caused by the virus. Andrographolide and its derivatives have been widely used for treating respiratory infections in China for decades. We have recently synthesized new andrographolide derivatives and found that some of the compounds including 14-alpha-lipoyl andrographolide (AL-1) have significant activity against bacterial infections with an unique mechanism of action. We report here the antiviral activity of AL-1 and other andrographolide drugs. AL-1 showed significant activity against influenza A viruses including the H5N1 avian influenza virus. The administration of AL-1 by oral gavage to mice infected with avian influenza A/Chicken/Guangdong /96 (H9N2), A/Duck/Guangdong/99 (H5N1), and human influenza A/PR/8/34 (H1N1) viruses greatly reduced the death rate, prolonged life, inhibited lung consolidation, and reduced viral titers in the lung. The most effective dosage of AL-1 in these studies ranged from 100 to 200 mg/kg/d, when administered twice daily for 7 d beginning 24 h before viral exposure. The LD(50) of AL-1 was 1243 mg/kg/d. AL-1 was effective against avian influenza A (H9N2 and H5N1) and human influenza A H1N1 viruses in vitro, with the 50% effective concentrations ranging from 7.2 to 15.2 microM and the selective indexes ranging from 51 to 109. Significant inhibition of viral adsorption onto red blood cells with minimum inhibitory concentrations ranging from 5.3 to 16.8 mM suggested that AL-1 was capable of directly interfering with viral hemagglutinin to block binding to cellular receptors. With potent antiviral activity and a potentially new mechanism of action, AL-1 may warrant further evaluation as a possible therapy for influenza.

The distribution characteristics of hexachlorobenzene (HCB) in plant and rhizosphere soil in contamination conduit, a nearby river and a cropland were studied and the impact factors were also discussed. The results are summarized as follows: the range of the HCB concentration in plant and rhizosphere soil in investigation area were respectively from 4.45 microg x kg(-1) to 1,189.89 microg x kg(-1) (dw) and from 27.93 microg x kg(-1) to 3,480.71 microg x kg(-1) (dw). Higher enrichment of HCB in woodplant than herbs due to higher fat concentration in woodplant in the contamination conduit and the rich concentrtion factor of woodplant and herbs were 0.41-2.55 and 0.01-1.34. The range of HCB concentrations in plants in nearby croplands was significantly wide (4.45-333.1 microg x kg(-1)) while HCB concentrations in different parts of plant were various, e.g. HCB concentrations in fruit, root and shoot of taro were 318.77 microg x kg(-1), 281.02 microg x kg(-1) and 10.94 microg x kg(-1). There was a remarkable positive relation between the concentrations of HCB in plant and fat concentration of plant while no relativity between the concentrations of HCB in plant and those in ground soils in the contamination conduit and cropland. The concentration levels of HCB in plant and rhizosphere soil in river were dramatically decreased with increasing distance from contaminated conduit. There was a remarkable positive relation between the concentrations of HCB in plant and those in ground soils but no relation between concentrations of HCB in plant and fat concentration of plant in river. The distribution characteristics of HCB in plants were influenced by contaminated levels, fat concentration and Partition-transfer model.

To study the releasing properties and tissue-targeting characteristics of methotrexate-loaded red blood cells (MTX-RBCs), pharmacokinetics and tissue distributions of intravenous injected MTX-RBCs and free MTX were compared. MTX-RBCs were made from rat erythrocytes using a hypertonic method. After i.v. injection of MTX-RBCs or free MTX to rats, both plasma and tissue homogenate samples at each time-point were collected and analyzed by RP-HPLC. From this data, pharmacokinetic parameters and tissue distributions of MTX were obtained. MTX-RBCs were successfully produced by the hypertonic method. After i.v. injection, MTX-RBCs displayed more than three times longer half-life and MRT than free MTX, and the velocity of MTX clearance from plasma was much slower. The ratio of area under the concentration time curve (AUC)(tissue) to AUC(plasma) in the liver was clearly higher than that for other organs after MTX-RBCs administration; MRT in the liver was also longer. This study has demonstrated that the hypertonic method for making MTX-RBCs has led to a preparation with slow release properties as well as liver-targeting characteristics in rats. This approach offers considerable potential for the treatment of tumors in liver, which would merit further investigation.

We report the cloning and characterization of TOX3, a high mobility group box protein involved in mediating calcium-dependent transcription. TOX3 was identified as a calcium-dependent transactivator using the Transactivator Trap screen. We find that TOX3 interacts with both cAMP response element (CRE)-binding protein (CREB) and CREB-binding protein (CBP), and knockdown of the endogenous TOX3 by RNAi leads to significant reduction of calcium-induced c-fos expression and complete inhibition of calcium activation of the c-fos promoter. The effects of TOX3 on calcium-dependent transcription require the CRE elements. These observations identify TOX3 as an important regulator of calcium-dependent transcription and suggest that TOX3 exerts its effect on CRE-mediated transcription via its association with the CREB-CBP complex.

Purpose: To clarify the prognostic value of molecular diagnosis of SLN metastases in cervical cancer using SCCA., Experimental Design: All SLNs and primary tumors, part of non-SLNs, were harvested from 36 patients with cervical cancer. Expression levels of SCCA, cytokeratin 19 (CK19), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA in 178 samples (29 primary tumors, 5 histologic positive nodes, 60 histologic negative SLNs, 69 non-SLNs, and 15 normal nodes) were assessed by quantitative reverse transcription-PCR assay. The quantitative value of SCCA or CK19 mRNA was described as each value relative to GAPDH mRNA. The cutoff value was set at the upper limit of the quantitative value of nodes from noncancer patients, and those above this value constituted the molecular metastasis group., Results: The SCCA mRNA expression values were more than 1 x 10(3) in 28 primary tumors and all histologic positive nodes, and its expression levels in SLNs were higher than in non-SLNs. SLNs from patients with adverse prognostic features had higher SCCA mRNA expression levels. Four histologic negative SLNs were diagnosed molecular metastases based on SCCA mRNA. Two cases with histologically uninvolved pelvic nodes recurred. Survival analysis indicates that molecular lymphatic metastasis based on elevated SCCA mRNA level is the best predictor of recurrence. However, CK19 is not a suitable marker due to its low specificity and relative higher baseline expression in normal nodes., Conclusions: SCCA mRNA levels for molecular diagnosis of SLN metastases in cervical cancer more accurately identifies patients at risk for recurrence than the routine histology does.

Objective: The human socio-economic development depends on the planet's natural capital. Humans have had a considerable impact on the earth, such as resources depression and environment deterioration. The objective of this study was to assess the impact of socio-economic development on the ecological environment of Wuhan, Hubei Province, China, during the general planning period 2006-2020., Methods: Support vector machine (SVM) model was constructed to simulate the process of eco-economic system of Wuhan. Socio-economic factors of urban total ecological footprint (TEF) were selected by partial least squares (PLS) and leave-one-out cross validation (LOOCV). Historical data of socio-economic factors as inputs, and corresponding historical data of TEF as target outputs, were presented to identify and validate the SVM model. When predicted input data after 2005 were presented to trained model as generalization sets, TEFs of 2005, 2006,..., till 2020 were simulated as output in succession., Results: Up to 2020, the district would have suffered an accumulative TEF of 28.374 million gha, which was over 1.5 times that of 2004 and nearly 3 times that of 1988. The per capita EF would be up to 3.019 gha in 2020., Conclusions: The simulation indicated that although the increase rate of GDP would be restricted in a lower level during the general planning period, urban ecological environment burden could not respond to the socio-economic circumstances promptly. SVM provides tools for dynamic assessment of regional eco-environment. However, there still exist limitations and disadvantages in the model. We believe that the next logical step in deriving better dynamic models of ecosystem is to integrate SVM and other algorithms or technologies.

A randomized single-blind trial was conducted to evaluate the efficacy of short-term abdominal acupuncture as a novel treatment for insomnia in Chinese women. Forty-four patients between the ages of 22 and 56 were randomly assigned to an acupuncture (n = 23) and a medication group (n = 21). The acupuncture group received abdominal acupuncture once a day for the first three days and once every three days for the remaining 11 days. In addition, every subject in acupuncture group also received a placebo pill once daily. Abdominal acupuncture was administered according to a standardized protocol involving four master and four adjunctive acupoints: Zhongwan (CV 12), Xiawan (CV 10), Guanyuan (CV 4), and Qihai (CV 6); bilateral Shangqu (KI 17), Huaroumen (ST 24), Xiafengshidian, and Qipang. Subjects in the medication group were treated with sham acupuncture at the same time as the acupuncture group and received estazolam once a day. The outcome measure was the Leeds Sleep Evaluation Questionnaire (LSEQ), administered before and after the trial. Subjects who received abdominal acupuncture lowered their LSEQ scores by an average of 26.32 points (95% CI: 37.34, 15.30). After controlling for potential confounding factors, the effect of abdominal acupuncture in relieving insomnia was still statistically significant. Results indicate that short-term abdominal acupuncture is more effective than pharmacological treatment for relieving insomnia in adult women and has few adverse effects.

Soy odor is a major concern in the consumption of soymilk by Western consumers. The objectives of this study were to determine selected soy odor compounds as affected by soybean materials and direct steam injection and to determine if the steam-injection method affected overall cooked soy aroma of the soymilk and compared to the soymilk cooked by a traditional indirect method. Five varieties of soybeans with or without lipoxygenases were processed by direct-steam injection for up to 20 min at 100 degrees C. Eight selected odor compounds were analyzed by gas chromatography after extraction using a solid-phase micro-extraction (SPME) method. Hexanal, hexanol, 1-octen-3-ol, 1-octen-3-one, and trans-2-nonenal decreased whereas 2-pentylfuran and trans-2, trans-4-decadienal increased by boiling up to 20 min. The results showed that soybean variety was a significant factor to affect odor compositions. Direct steam injection cooked soymilk resulted in lower odor contents than a traditional indirect cooking method. The advantage of having a low odor composition in the heated soymilk products made from lipoxygenases-null soybean varieties could be attained by using normal soybean materials with direct steam injection.

Epidemiological studies demonstrated that the consumption of dietary antioxidant was associated with the prevention of atherosclerosis. The aim of this study was to investigate the antioxidant activities of the hydrophilic extracts from 9 selected legumes based on copper-induced human LDL oxidation model in vitro. The antioxidant activities were assessed on the basis of the formation of conjugated dienes (lag time of oxidation) and thiobarbituric acid reactive substances (TBARS) as the early and later stage markers of LDL oxidation. The results showed that the extracts of black beans (Phaseolus vulgaris L.), lentils (Lens culinaris), black soybeans (Glycine max), and red kidney beans (Phaseolus vulgaris L.) had significant (P < 0.05) longer LDL oxidation lag times (128.8, 124.2, 107.7, and 111.1 min, respectively) than the LDL control group (94.9 min). No significant lag-time lengthening was observed in other tested legume extracts. On the other hand, black beans, lentils, black soybeans, red kidney beans, and pinto beans exhibited higher antioxidant capacities (Trolox equivalents) than yellow peas, green peas, chickpea, and yellow soybeans in both LDL-conjugated dienes assay and LDL-TBARS assay. Meanwhile, the antioxidant activities of these legumes against LDL-lipid peroxidation in the above assays were found to correlate very significantly (P < 0.01) with their phenolic substances, and DPPH radical scavenging activity and ORAC (oxygen radical absorbance capacity). These results suggest that consuming black beans, lentils, black soybeans, and red kidney beans may have potential in preventing the development of atherosclerosis from the perspective of inhibiting LDL oxidation.

Objective: To evaluate the feasibility of sentinel lymph node (SLN) detection in patients with cervical cancer using the low-cost methylene blue dye and to optimize the application procedure., Patients and Methods: Patients with stage Ib(1)-IIa cervical cancer and subjected to abdominal radical abdominal hysterectomy and pelvic lymphadenectomy were enrolled. Methylene blue, 2-4 ml, was injected into the cervical peritumoral area in 77 cases (4 ml patent blue in the other four cases) 10-360 min before the incision, and surgically removed lymph nodes were examined for the blue lymph nodes that were considered as SLNs., Results: High SLN detection rate was successfully achieved when 4 ml of methylene blue was applied (93.9%, 46/49). Bilaterally SLN detection rate was significantly higher (78.1% vs. 47.1% P=0.027) in cases when the timing of application was more than 60 min before surgery than those with timing no more than 30 min. The blue color of methylene blue-stained SLNs sustained both in vivo and ex vivo, compared with the gradually faded blue color of patent blue that detected in 3 of 4 cases unilaterally. In the total of 112 dissected sides, the most common location of SLNs was the obturator basin (65.2%, 73/112), followed by external iliac area (30.4%, 34/112) and internal iliac area (26.8%, 30/112). Three patients who gave false negative results all had enlarged nodes., Conclusion: Methylene blue is an effective tracer to detect SLNs in patients with early stage cervical cancer. The ideal dose and timing of methylene blue application are 4 ml and 60-90 min prior surgery, respectively.

Objective: To compare the combined FDG PET/CT with PET alone in the detection of lymph node metastasis (LNM) for esophageal carcinoma patient., Methods: From November 2003 to August 2005, 35 patients with esophageal carcinoma underwent FDG PET/CT before esophagectomy and lymph node (LN) dissection. The patients who had history of previous anticancer treatment or diabetes mellitus and inflammatory lung diseases as well as being inoperable for medical reasons were excluded. The results of LNM detection by PET/CT and PET alone were compared with pathological results., Results: Twenty-nine men and 6 women were eligible for this study, with a mean age of 57 years (range: 40 to 72 years). Of these 35 patients, 3 had lesion at the upper third thoracic esophagus, 22 at the middle third and 10 at the lower third. All patients underwent surgical resection successfully. Twenty-five patients and 65 out of 313 excised nodal groups were found to have metastases by pathological examination. The true positive and true negative LNM interpretation reached 61 and 229 LN groups on PET/CT versus 53 and 217 LN groups on PET alone. False negative LNM interpretation was found in 12 LN groups on PET alone, and 8 of them were corrected by PET/CT including one cervical LN, 4 paraesophageal LNs, one left gastric arterial LN, one left gastric cardia LN, one lesser gastric curve LN. False positive interpretations on PET alone were found in 31 LN groups, 12 of them were corrected by PET/CT which included 9 false-positive interpretations due to physical tracer uptake (2 in the cervical region and 7 in gastrointestinal tract ) and 3 lesions with heterogeneous tracer uptake in the primary tumor. The sensitivity, specificity and accuracy of LNM detection by PET alone was 81.54% (53/65), 87.50% (217/248), and 86.26% (270/313), whereas by PET/CT, which was 93.85% (61/65), 91.24% (229/248) and 92.65% (290/313), respectively. There were statistically significant differences in sensitivity and accuracy of LNM detection between PET/CT and PET alone (0.033 and 0.009)., Conclusion: Compared with FDG PET alone, FDG PET combined with CT can improve the sensitivity and accuracy in detection of lymph node metastasis in esophageal carcinoma.

The objective of this study was to characterize the phenolic compounds and antioxidant activities of U.S.-produced cool season legumes. A total of 33 cool season legume samples were selected. Some common beans and soybeans were included for comparisons. Total phenolic content (TPC), total flavonoid content (TFC), and condensed tannin content (CTC) were analyzed. Ferric reducing antioxidant power (FRAP), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, and oxygen radical absorbance capacity (ORAC) were used for analyzing antioxidant properties. Color of the legume flour and the seed coat was also analyzed. TPC, TFC, CTC, FRAP, DPPH, and ORAC values of legumes were significantly different not only between classes but also among samples within each class. Among cool season legume classes, lentils possessed the highest concentrations of the phenolic compounds and antioxidant activities. Colored common beans and black soybeans exhibited higher TPC, TFC, CTC, FRAP, DPPH, and ORAC values than those of yellow peas, green peas, and chickpeas. Antioxidant activities (FRAP, DPPH, and ORAC) were strongly correlated (r= 0.96, 0.94, and 0.89, respectively, P < 0.01) with TPC. TPC and ORAC were moderately correlated (P < 0.01) with either the seed hull surface color or the flour color.

Objective: To study the anti-inflammatory effects of Fu' an oral liquid ( FAOL)., Methods: The three experimental groups were administrated with FAOL of low, medium and high dosages, while the positive groups were given with aspirin and dexamethasone, and the control group was fed with distilled water. The effects of FAOL on the mice and rats were observed., Results: FAOL could evidently inhibit the increase of celiac capillary permeability caused by acetic acid in mice and reduce the swelling of adjuvant arthritis in rats induced by injecting methanal and egg serum. It could also inhibit the proliferation of granulation tissue induced by cotton-pellet in rats., Conclusion: FAOL has certain anti-inflammatory effects on acute and chronic inflammation.

This study investigated the electrokinetic (EK) behavior of multiple chlorobenzenes, including 1,2,3,4-tetrachlorobenzene (TeCB), 1,2,4,5-tetrachlorobenzene (i-TeCB), and 1,2,3-trichlorobenzene (TCB) in contaminated clayed soils. The effect of beta-cyclodextrin (beta-CD) on the EK removal of the chlorobenzenes was studied. The largest removal was obtained when Na2CO3/NaHCO3 buffer was used as anodic purging solution without beta-CD. The removal efficiencies were related to the aqueous solubilities of chlorobenzenes. With the same cumulative electroosmotic flow, greater solubility led to higher removal efficiency. The addition of beta-CD inhibited the EK removal efficiency of all chlorobenzenes. The inhibition increased with the increase of beta-CD concentration. With the same beta-CD concentration, the inhibition increased with the rise of electric potential. It was found that the inclusion compounds between beta-CD and chlorobenzenzes were less soluble than chlorobenzenes. The formation of the less soluble inclusion compounds reduced the aqueous solubility of chlorobenzenes and led to the partial immobilization of the chlorobenzenes that desorbed from soil. It was feasible to use the EK technology to remove chlorobenzenes in contaminated soils using water as the anodic flushing solution. The addition of beta-CD was not recommended for the EK removal of chlorobenzenes.

1. The Aim Of The Present Study Was To Test The Efficacy Of The Novel Endothelin (Et) Receptor Antagonist CPU0507 In Treating Rat Hypoxic Pulmonary Hypertension (Ph) In Vivo And In Vitro And To Explore The Role Of The Et-1 System In The Disease. 2. Male Sprague-dawley Rats (220 +/- 20 G) Were Divided Into Four Groups: (I) Control; (Ii) Untreated Hypoxic (28 Days Hypoxia); (Iii) Hypoxic Rats Treated In The Last 5 Days Of Hypoxia With Nifedipine(5 Mg/kg Per Day, P.o.); And (Iv) Hypoxic Rats Treated In The Last 5 Days Of Hypoxia With CPU0507 (20 Mg/kg Per Day, S.c.). Effects Of Treatments On Haemodynamics And Biochemical Data, As Well As Functional Assessments Of The Isolated Pulmonary Artery, Were Determined In Vivo And In Vitro. 3. It Was Found That CPU0507 Reduced The Elevated Pulmonary Arterial Pressure And Right Heart Weight Index And Restored Abnormalities In Nitric Oxide (No), Malondialdehyde And No Synthase (Nos) In The Serum And Superoxide Dismutase, Hydroxyproline And Nos In Pulmonary Homogenates. In Addition, CPU0507 Restored Altered Pulmonary Vasoconstrictor And Vasodilator Responses. Vascular Constriction And Dilatation Of Untreated Pulmonary Arteries Were Reverted Effectively Towards Normal Following Exposure Of Artery Rings To CPU0507 In Vitro. 4. In Conclusion, The Results Indicate That Hypoxic Ph Is Relieved Significantly By CPU0507 In Vivo And In Vitro And The Effects Are Presumed To Be Mediated By Suppression Of The Et-reactive Oxygen Species Axis.

Background & Objective: The prognosis of platinum-sensitive ovarian cancer patients was better than that of chemoresistant ones. However, platinum-sensitive ovarian cancer patients still have a high recurrence rate, which affects their prognosis. This study was designed to analyze clinical features and recurrence risk factors of platinum-sensitive epithelial ovarian cancer., Methods: Factors that might relate to recurrence of 90 platinum-sensitive epithelial ovarian cancer patients, admitted in Cancer Center of Sun Yat-sen University from 1993 to 1999, with complete remission of more than 6 months, were assessed. Univariate analysis was performed using Chi(2) test; while multivariate analysis was carried out using Cox proportional hazard model., Results: Among the 90 patients, 36(40.0%) relapsed with the median recurrence-free interval of 20 months. Pelvic cavity (18/36, 50.0%) was the most frequently involved. The 3-and 5-year survival rates of all patients were 79.6% and 69.5%; while those of the recurrent ones were 62.3% and 39.6%. Univariate analysis showed that the early FIGO stage group, mucinous type group, and no neoadjuvant chemotherapy group had lower recurrence rates than advanced FIGO stage group, non-mucinous type group, and neoadjuvant chemotherapy group, respectively (P=0.001, P=0.002, and P=0.025). Cox multivariate analysis showed that only FIGO stage was the independent risk factor of recurrence of ovarian cancer (risk ratio=1.771, P=0.003). There was no significant difference in recurrence rate between CBP and other postoperative chemotherapy regimen groups. More cycles of chemotherapy could not reduce the recurrence rate., Conclusion: Since FIGO stage is an independent recurrence risk factor of platinum-sensitive epithelial ovarian cancer patients, early diagnosis is the key point to decrease the recurrence rate.

Background & Objective: Metastasis suppressor gene KAI1/CD82 plays an important role in infiltration and metastasis of several types of human cancers, while the researches on its relation with cervical carcinoma are far from adequate now. Therefore, immunohistochemical techniques were employed in this study to determine the expression of KAI1 gene, and explore its clinical significance in cervical squamous cell carcinoma., Methods: SP immunohistochemistry was used to detect the expression of KAI1 gene in 99 specimens of cervical squamous cell carcinoma, 25 specimens of cervical intraepithelial neoplasm (CIN) II-III, and 18 specimens of normal cervix. Correlations of expression of KAI1 gene to clinicopathologic factors, and prognosis of cervical squamous cell carcinoma were statistically analyzed., Results: The rates of negative, weak, moderate, and strong expression of KAI1 in cervical squamous cell carcinoma were 52.5% (52/99), 16.2% (16/99), 15.2% (15/99), and 16.2% (16/99), respectively, significantly lower than those in normal cervix, and CIN II-III (P=0.000). Expression of KAI1 has no correlation with FIGO stage, age, pelvic lymph node metastasis, tumor histological grade, depth of cervical infiltration, serum squamous cell carcinoma antigen (SCC) level, tumor size, and gross type of cervical lesion (P>0.05). Both univariate and multivariate analyses showed expression of KAI1 has no correlation with prognosis of cervical squamous cell carcinoma (P>0.05)., Conclusion: KAI1 gene may be an early event in development of cervical cancer, and has no correlation with prognosis of cervical squamous cell carcinoma.

Background & Objective: Study of sentinel lymph node(SLN) in cervical cancer has been initiated since recent years, and there are still a lot of unknown factors about SLN identification in cervical cancer. This study was to investigate influential factors of identifying SLN with methylene blue in cervical cancer., Methods: For 41 patients with cervical cancer enrolled from Jun. 2002 to May 2003, 2-4 ml of methylene blue was injected into cervix at 4-6 sites around the tumor about 90-400 min before operation. The blue-dyed lymph node (BDLN) was considered as SLN. HE staining in step sections, and immunohistochemistry were applied to detect SLN. The influential factors of using methylene blue to detect SLN in cervical cancer were assessed based on the identification rate of SLN, and its false negative rate., Results: SLNs were detected in 31 of 41(75.6%) patients with cervical cancer of stage Ib1-IIb. A total of 85 SLNs were identified,and most frequently located in obturator fossa. SLNs were successfully detected in 20 of 23 (87.0%) patients who had no preoperative radiotherapy or chemotherapy, while in only 11 of 18 (61.1%) patients who had preoperative radiotherapy and/or chemotherapy. SLNs were detected in only 17 of 27(63.0%) patients who were injected with 2-3 ml of methylene blue, while in all of 14 patients whose dose was 3.4-4 ml. Eight patients were confirmed of lymph node metastases by pathology., Conclusions: The dose of methylene blue recommended to detect SLN in cervical cancer is 3-4 ml. SLN varies in different sites, but most frequently located in obturator fossa.

Objective: To evaluate the clinical effects of garlic plaster on the recurrent oral ulcer (ROU)., Methods: The garlic powder was made by special method and made into garlic plaster including 0.1% garlicin. The plaster was painted on the surface of ROU to examine its clinical effect. The patients were followed up for 1 to 4 years., Results: Of the 30 patients treated with garlic plaster, the complete effective rate was 83.3%, the partial effective rate was 16.7%, and the total effective rate was 100%. Fourteen patients did not recur 4 years after the treatment (46.7%)., Conclusion: Garlic has a broad clinical prospect due to its effective treatment on ROU.

Background & Objective: Vulvar Paget's disease is a rare disease. It has a relatively high misdiagnosis rate and there are still controversies regarding its treatment. The objective of this study was to investigate its clinical features and summarize the experience of the management of this disease in order to gain a better acknowledge of this rare disease and improve its cure rate., Methods: The clinical records of 8 cases of vulvar Paget's disease admitted in Cancer Center, Sun Yet-sen University from January 1964 to December 2001 were analyzed., Results: The average age of 8 cases was 66.5 years, and the mean time from the onset of the disease to diagnosis was 5 years. Pathologically, intraepithelial Paget's disease was the commonest (5/8), followed by invasive Paget's disease (2/8) and Paget's disease with underlying adenocarcinoma (1/8). Eight cases underwent altogether 10 times of surgery, and radical vulvectomy was the most frequently used procedure (6/10). One case of Paget's disease with underlying adenocarcinoma died 21 months after surgery and 1 case of intraepithelial Paget's disease died of other etiology, 5 cases achieved long-term disease free survival, 1 case developed recurrence., Conclusion: Surgery is the first choice for the patients of vulvar Paget's disease. For those patients whose lesions are extensive,or old aged,or not suitable for long-term follow-up, a radical vulvectomy is more preferable and a lower recurrence rate is expected.

Background & Objective: Recent researches manifested that down-regulation of p21(WAF1) had relationship with carcinogenesis and development in various tumors, but its association with epithelial ovarian cancer (EOC) was not clear. This study was designed to investigate the role of p21(WAF1) in the tumorigenesis and development of EOC and its relationship with p53 and proliferating cell nuclear antigen (PCNA) protein., Methods: Fifty-five EOC tissues, 32 benign ovarian tumor tissues, and 30 normal ovarian tissues were collected. Reverse transcription polymerase chain reaction (RT-PCR) was applied to determine the p21(WAF1)mRNA expression. Immunohistochemistry was applied to examine the protein expression of p21(WAF1), p53, and PCNA. The relationship between the expression of these markers and the clinicopathological characteristics, prognosis of the patients was analyzed., Results: The positive rates of p21(WAF1)mRNA in EOC, benign ovarian tumor, and normal ovary were 40%, 56.25%, and 73.33%, respectively (P=0.012). The positive rates of p21(WAF1) protein were 36.36%, 56.25%, and 80%, respectively (P=0.001). The positive expression rates of p21(WAF1)mRNA and its protein in EOC were lower than those of the other two groups, while the positive expression rates of p53 and PCNA protein in EOC were higher than those of the other two groups (P< 0.05). Expression of p21(WAF1)mRNA had positive relation to its protein, negative relation to PCNA protein, no relation to p53 protein, while expression of p21(WAF1) protein had negative relation to p53 and PCNA protein in EOC. Low-expression of p21(WAF1) protein was associated with advanced FIGO stage (P=0.032), but not with age, histological type, pathological grade, and remnant tumor (P >0.05). There was no relationship between p21(WAF1)mRNA and former parameters (P >0.05). Univariate analysis showed that the patients with low-expression of p21(WAF1)mRNA and p21(WAF1) protein had poor prognosis (P< 0.05)., Conclusion: p21(WAF1) is down-regulated in EOC. p21(WAF1) might be able to be used as a marker to predict the prognosis of patients with EOC.

Apoptosis is necessary for the development and maturation of Leydig cells. However, increased apoptosis results the decline of testosterone production, which may increase germ cell apoptosis and the possibility of infertility. There are several aspects contributing to Leydig cell apoptosis such as ethane dimethanesulphonate (EDS), glucocorticoid, developmental stage and some hormones including FSH, LH/hCG and testosterone. A number of genes are involved in the regulation of Leydig cells apoptosis. It was reported that SCF/c-kit, Bcl-2 and Bcl-xl inhibited the apoptosis while caspase-3, Fas, Bax and clusterine stimulated it.

OBJECTIVE: To investigate the feasibility of multiple loci detection in single cell by primer extension preamplification (PEP) followed by nest PCR. METHODS: Using PEP, the whole genomic DNA in single lymphocyte or single blastomere was amplified. In addition, CD17, nt-28 and linked ATTTT repeat for beta-thalassemia, F508 and linked GATT repeat for cystic fibrosis, DMD exon 17 and 48 for Duchenne muscular dystrophy, short tandem repeats of D18S51, D21S11 and D21S1411, and sex-determination gene SRY of the Y chromosome were all detected using nest-PCR from a small aliquot of the PEP reaction. RESULTS: The rate of successful single lymphocyte amplification was 89.5%(false positive 0.48%false negative 2.5%). The rate of successful single blastomere amplification was 85.56%(false positive 3%). CONCLUSION: The PEP technique followed by nest PCR analysis of single cell is very useful for simultaneous detection of multiple gene loci. It may be applicable for preimplantation genetic diagnosis.

A 5-year-old Caucasian male presented with early loss of multiple deciduous teeth. All the characteristics were consistent with the diagnosis of regional odontodysplasia (ROD). Significant initial findings included premature loss of multiple primary mandibular teeth and some malformed permanent teeth. The affected teeth showed hypoplastic enamel and dentin, short roots, and wide pulp chambers, and were localized in the mandibular dentition. Treatment objectives for this patient were to provide improved esthetics, restored chewing function, and space maintenance by the construction of a temporary prosthetic restoration. However, with limited tooth support and an unusual occlusal pattern, it is difficult to obtain satisfactory retention and esthetics with traditional prosthetic techniques. In this article we introduce an alternative method for fabricating a custom removable denture and discuss the prognosis of the malformed permanent dentition and further treatment plan.

Our previous immunofluorescence studies support the conclusion that the temporal appearance and subcellular distribution of TS28 (a marker of transverse (T) tubules and caveolae in adult skeletal muscle [Jorgensen, A. O., W. Arnold, A. C.-Y. Shen. S. Yuan, M. Gover, and K. P. Campbell, 1990, J. Cell Biol. 110:1173-1185]), correspond very closely to those of T-tubules forming de novo in developing rabbit skeletal muscle (Yuan, S., W. Arnold, and A. O. Jorgensen, 1990, J. Cell Biol. 110:1187-1198). To extend our morphological studies of the biogenesis of T-tubules and triads, the temporal appearance and subcellular distribution of the alpha 1-subunit of the 1,4-dihydropyridine receptor (a marker of the T-tubules and caveolae) was compared to (a) that of TS28; and (b) that of the ryanodine receptor (a marker of the junctional sarcoplasmic reticulum) in rabbit skeletal muscle cells developing in situ (day 19 of gestation to 10 d newborn) by double immunofluorescence labeling. The results presented show that the temporal appearance and relative subcellular distribution of the alpha 1-subunit of the 1,4-dihydropyridine receptor (alpha 1-DHPR) are distinct from those of TS28 at the onset of the biogenesis of T-tubules. Thus, in a particular developing myotube the alpha 1-DHPR appeared before TS28 (secondary myotubes; day 19-24 of gestation). Furthermore, the alpha 1-DHPR was distributed in discrete foci at the outer zone of the cytosol, while TS28 was confined to foci and rod-like structures at the cell periphery. As development proceeded (primary myotubes; day 24 of gestation) approximately 50% of the foci were positively labeled for both TS28 and the alpha 1-DHPR, while approximately 20 and 30% of the foci were uniquely labeled for TS28 and the alpha 1-DHPR, respectively. The foci labeled for both TS28 and the alpha 1-DHPR and the foci uniquely labeled for TS28 were generally confined to the cell periphery, while the foci uniquely labeled for the alpha 1-DHPR were mostly confined to the outer zone of the cytosol. 1-2 d after birth, TS28 was distributed in a chickenwire-like network throughout the cytosol, while the alpha 1-DHPR was confined to cytosolic foci. In contrast, the temporal appearance and subcellular distribution of the alpha 1-DHPR and the ryanodine receptor were very similar, if not identical, throughout all the stages of the de novo biogenesis of T-tubules and triads examined.(ABSTRACT TRUNCATED AT 400 WORDS)

Novel proteins unique to either transverse tubules (TS28) or the sarcolemma (SL50) have been identified and characterized, and their in situ distribution in rabbit skeletal muscle has been determined using monoclonal antibodies. TS28, defined by mAb IXE112, was shown to have an apparent relative molecular mass of 28,000 D. Biochemical studies showed that TS28 is a minor membrane protein in isolated transverse tubular vesicles. Immunofluorescence and immunoelectron microscopical studies showed that TS28 is localized to the transverse tubules and in some subsarcolemmal vesicles possibly corresponding to the subgroup of caveolae connecting the transverse tubules with the sarcolemma. In contrast, TS28 is absent from the lateral portion of the sarcolemma. Immunofluorescence studies also showed that TS28 is more densely distributed in type II (fast) than in type I (slow) myofibers. Although TS28 and the 1,4-dihydropyridine receptor are both localized to transverse tubules and subsarcolemmal vesicles, TS28 is not a wheat germ agglutinin (WGA)-binding glycoprotein and does not appear to copurify with the 1,4-dihydropyridine receptor after detergent solubilization of transverse tubular membranes. SL50, defined by mAb IVD31, was shown to have an apparent relative molecular mass of 50,000 D. Biochemical studies showed that SL50 is not related to the 52,000-D (beta subunit) of the dihydropyridine receptor but does bind to WGA-Sepharose. Immunofluorescence labeling imaged by standard and confocal microscopy showed that SL50 is associated with the sarcolemma but apparently absent from the transverse tubules. Immunofluorescence labeling also showed that the density of SL50 in type II (fast) myofibers is indistinguishable from that of type I (slow) myofibers. The functions of TS28 and SL50 are presently unknown. However, the distinct distribution of TS28 to the transverse tubules and subsarcolemmal vesicles as determined by immunocytochemical labeling suggests that TS28 may be directly involved in excitation-contraction coupling. Our results demonstrate that, although transverse tubules are continuous with the sarcolemma, each of these membranes contain one or more unique proteins, thus supporting the idea that they each have a distinct protein composition.

Activation of protein kinase C has been implicated in the regulation of a variety of cellular reactions. Although we, and others, have found protein kinase C and its substrate proteins to be present in both membrane and cytosolic fractions in the heart, the physiologic role of this kinase in the regulation of cardiac functions remains unknown. In the present study, we found that in isolated perfused rat heart, administration of phorbol esters 4 beta-phorbol 12,13-dibutyrate(PDBu) and 12-0-tetradecanoylphorbol 13-acetate(TPA), which are specific activators of protein kinase C, produced marked dose-dependent negative changes in inotropy and chronotropy. A dose-dependent decrease in coronary flow was also observed. The diacylglycerol analogues, 1,2-oleoylacetyl-glycerol and 1,2-dioctanoylglycerol, produced similar effects as the active phorbol esters on these isolated perfused hearts. An inactive analogue of phorbol ester, 4 alpha-phorbol, failed to produce any effect. Protein kinase C activity in both membrane and cytosolic fractions prepared from rat heart could be activated by TPA and PDBu at the same concentration range as used in the experiments with perfused hearts. Following perfusion of the hearts with PDBu, a rapid translocation of protein kinase C from cytosolic to membrane fractions was also observed. Our findings provide the first direct evidence that protein kinase C may play a potentially important role in the regulation of cardiac functions.