Objective To explore the effects of alcoholic extracts of traditional Chinese medicines on the post-translational processing and trafficking of tyrosinase.Methods Human YUGEN8 amelanotic melanoma cells were grown in vitro;the cells were incubated with one of the seven traditional Chinese medicines,including Rhizoma Chuanxiong and psoralen.Protein analysis with Western blot,enzymolysis with endoglycosidase H (Endo H),and subcellular localization with laser confocal microscopy were per- formed.The expression,maturity and export from endoplasmic reticulum (ER) of tyrosinase in the treated cells were compared with those in the untreated controls.Results Compared with controls,an approximate- ly 80-kDa,Endo H-resistant tyrosinase doublet,which represented mature glycoform of tyrosinase,was in- creased in melanocytes treated with Semen Cuscutae,and in those treated with Semen Persicae.Within those cells,tyrosinase was distributed outside ER resident protein calnexin.Conclusion Both Semen Cus- cutae and Semen Persicae could induce tyrosinase maturation,stability and export from ER to distal site.

BACKGROUNDDifferential diagnosis of isolated calf muscle vein thrombosis (ICMVT) and gastrocnemius hematoma is essential for early identification of deep vein thrombosis (DVT). This study aimed to investigate the diagnostic value of high-frequency color Doppler ultrasound for differential diagnosis of ICMVT and gastrocnemius hematoma.

METHODSA retrospective case series of 35 ICMVT (M:F, 21:14; mean age (64.5 ± 10.6) years) and 23 gastrocnemius hematoma (M:F, 16:7; mean age (75.4 ± 11.8) years) patients with bilateral/unilateral lower limb pain was conducted between January 2006 and September 2012. Characteristics and the morphology of high-frequency color Doppler ultrasonography of the lower limb deep vein, great saphenous vein, calf muscles, skin, and soft tissue were examined.

RESULTSICMVT hypoechoic signals were characterized by long, tube-like masses on longitudinal sections and oval masses on transverse sections, with apparent muscle thrombosis boundaries, distal and proximal venous connections, and, often, lower limb DVT. Gastrocnemius hematoma hypoechoic signals were characterized by large volumes, enhanced posterior hematoma echo, hyperechoic muscle boundaries, no hematoma blood flow, and no DVT, and clear differences in trauma/exercise- and oral anticoagulant-induced hematomas were readily apparent. According to the measurement, the ratio of long diameter/transverse diameter (D/T) in ICMVT patients was about less than 2.0, whereas in gastrocnemius hematoma patients the ratio was more than 2.0. Early stage isoechoic and hypoechoic signals were detected with gradually increasing ovular anechoic areas. Partial muscle fibers in the hematoma due to muscle fractures were apparent.

CONCLUSIONHigh-frequency color Doppler ultrasound was found to be a sensitive and reliable method for differential diagnosis of ICMVT and gastrocnemius hematoma due to trauma and exercise or prolonged oral anticoagulant use.

Aged ; Diagnosis, Differential ; Female ; Humans ; Leg ; diagnostic imaging ; Male ; Middle Aged ; Muscle, Skeletal ; diagnostic imaging ; Retrospective Studies ; Ultrasonography ; Venous Thrombosis ; diagnostic imaging

OBJECTIVETo investigate the epidemiological characteristics of human papillomavirus (HPV) infection in men attending a sexually transmitted diseases (STD) clinic in Hangzhou area.

METHODSMale subjects (n=375) aged 18-70 years, attending the STD clinic were recruited. Urethral swabs were assessed for HPV DNA using polymerase chain reaction (PCR) with the consensus primers MY09/11. HPV genotypes of positive PCR products were determined by restriction fragment length polymorphisms and direct sequence analysis.

RESULTSOf the 375 swabs collected, 305 (81.3%) yielded sufficient DNA for the subsequent HPV analysis. Among the 305 subjects, the prevalence of HPV was 13.8%. Nononcogenic HPV types were found in 8.5% (26/305) of subjects, oncogenic types in 4.3% (13/305), and multiple types in 1.0% (3/305). The prevalence of HPV infection was higher in subjects from urban area than in those from rural area (P < 0.05). The prevalence was also higher in those who received fewer years of education (P < 0.05) and those who had more sex partners (P < 0.05).

CONCLUSIONSHPV infection among men at high risk is not uncommon. The detection rate of HPV DNA is significantly related to some sociodemographic factors, such as residence, educational level and the number of sex partners.

Adolescent ; Adult ; Aged ; Ambulatory Care Facilities ; China ; epidemiology ; Humans ; Male ; Middle Aged ; Papillomaviridae ; classification ; genetics ; isolation & purification ; Papillomavirus Infections ; diagnosis ; epidemiology ; virology ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sexually Transmitted Diseases ; prevention & control

OBJECTIVETo study the prevalence of primary HIV drug resistance in antiretroviral therapy (ART) areas of Henan province.

METHODSA total of 121 drug-naive long-term infected individuals and 154 patients with newly diagnosed from January 2011 to March 2012 were recruited, the questionnaires were surveyed and whole blood were collected to analyze the CD4(+)T cell counts and viral load. In-house method for genotypic resistance test was determined in those with viral load > 1000 copies/ml samples, the differences of demographic characteristics, immunological parameters and primary drug resistance were compared between the two groups.

RESULTSA total of 121 cases of long-term individuals who had infected (12.50 ± 3.21) years were mainly previous paid blood donors, and the age was (46.61 ± 9.32) years old. The infection route of the newly diagnosed were diversity, including blood, sexual transmission and others, the cases were 73, 73, 8, respectively, the confirmatory year was (0.91 ± 0.28) years, and average age was (22.21 ± 3.11) years old. The difference were statistically significant in the route of transmission, age and infection time from demographic analysis of the two groups (P < 0.05). The absolute M(P(25)-P(75)) counts of CD4(+)T lymphocytes of long-term group was 322 (217 - 422) cell/µl, which was lower than the newly diagnosed was 434(308 - 578) cell/µl (P < 0.05), and viral load was 4.0 (2.96 - 4.64) copies/ml, 3.77 (2.94 - 4.53) copies/ml, the difference was not significant (P > 0.05). The prevalence of primary drug resistance in long-term group and newly diagnosed was 5.79% (7/121), 9.09% (14/154), respectively, and the difference was statistically different (P < 0.05), and one PI-resistant strain was found in the newly diagnosed group.

CONCLUSIONThe primary drug resistant strains in untreated patients were found in Henan province of ART areas, and there was difference in degree of resistance between long-term infected individuals and newly diagnosed.

Adult ; Anti-HIV Agents ; pharmacology ; therapeutic use ; China ; epidemiology ; Drug Resistance, Viral ; Female ; HIV Infections ; drug therapy ; epidemiology ; virology ; Humans ; Male ; Middle Aged ; Viral Load

OBJECTIVETo investigate the enhanced predictive activity of preoperative plasma osteopontin (OPN) level in combination with intercellular adhesion molecule-1 (ICAM-1) for recurrence and prognosis of patients after resection of hepatocellular carcinoma (HCC).

METHODSA total of 75 patients received liver resection for HCC from August 2001 to December 2001 in authors' institute were enrolled in this study. The preoperative plasma levels of OPN and ICAM-1 were detected by ELISA, and the association of them combination with the recurrence and prognosis of HCC patients was analyzed.

RESULTSOPN and ICAM-1 could be detected in all of the plasma samples of the tested patients. A significantly higher OPN level and ICAM-1 level were found in plasma of patients who were found to have HCC recurrence during the follow-up time compared with those without recurrence (210.40 vs. 154.86 ng/ml, P = 0.001; 1011.23 vs. 747.49 ng/ml, P = 0.027). A significant difference of OS and DFS were found in different subgroups with higher or lower level of OPN (625 vs. 808 days, P = 0.0006; 433 vs. 674 days, P = 0.0003); and a similar situation was found in patients of high- and low- ICAM-1 levels (651 vs. 794 days, P = 0.0269; 489 vs. 642 days, P = 0.0248). The 2-year recurrence rates of the patients with higher and lower plasma levels of both OPN and ICAM-1 were 87.50% and 28.00% (P < 0.001), respectively; and the 2-year OS rates were 37.50% and 88.00% (P = 0.001), and the 2-year DFS rates were 12.50%, and 76.00 (P = 0.001), respectively.

CONCLUSIONSThe evaluation of preoperative plasma level of OPN or ICAM-1 may be helpful to predict the recurrence and prognosis of HCC patients in advance. The assessment of OPN level in combination with ICAM-1 could stratify patients into groups with different potentials of HCC recurrence and different outcomes more accurately than OPN or ICAM-1 individually.

Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; blood ; Carcinoma, Hepatocellular ; diagnosis ; Female ; Humans ; Intercellular Adhesion Molecule-1 ; blood ; Liver Neoplasms ; diagnosis ; Male ; Middle Aged ; Osteopontin ; blood ; Prognosis

OBJECTIVEIn order to seek the functional evidence that there could be metastatsis suppressor gene for liver cancer on human chromosomes, the objective of this study is to establish a method of microcell mediated chromosome transfer (MMCT).

METHODSHuman chromosome 8 randomly marked with neo gene was introduced into highly metastatic rat liver cancer C5F cell line by treating the single human chromosome donor cells with sequential steps of micronucleation, enucleation and microcell fusion. Double selections of G418 and HAT were applied to screen positive microcell hybrids, which were cloned by single cell isolation. Microcell hybrid clones were confirmed by STS-PCR and WCP-FISH.

RESULTSMicrocell hybrids resistant to HAT and G418 were obtained, from which 15 clones were obtained by single-cell isolation cloning. STS-PCR and WCP-FISH proved that human chromosome 8 had been successfully introduced into rat liver cancer cell line C5F. The human chromosome 8 introduced into C5F was found to have random loss of chromosome fragments by STS-PCR and consistent recombination with rat chromosome by WCP-FISH.

CONCLUSIONThe successfulls introduction of human chromosome into highly metastatic rat liver cancer cell line has established the technical basis for functional localization of metastasis suppressor gene(s) for liver cancer on human chromosomes.

Animals ; Cell Line, Tumor ; Chromosome Mapping ; methods ; Chromosomes, Human, Pair 8 ; genetics ; Genes, Tumor Suppressor ; Genetic Techniques ; Humans ; In Situ Hybridization, Fluorescence ; Liver Neoplasms ; genetics ; pathology ; Polymerase Chain Reaction ; Rats

OBJECTIVESTo detect the loss of heterozygosity (LOH) of circulating DNA in the plasma of patients with hepatocellular carcinoma (HCC), and to assess its potential as a clinical predictive marker.

METHODSThree high-polymorphic microsatellite markers D8S277, D8S298 and D8S1771 located at chromosome 8p were selected to detect LOH in plasma DNA of 62 HCC patients. The associations between LOH and its clinicopathological features, including HBsAg, liver cirrhosis, serum AFP level, tumor size, tumor cell differentiation, and intrahepatic metastasis were also examined.

RESULTSIn plasma DNA of the 62 HCC patients, LOH was found at one or several loci in 36 (58.1%), and heterozygosity at D8S277, D8S298, and D8S1771 loci was 74.2% (46/62), 75.8% (47/62), and 69.4% (43/62), respectively. LOH frequency at D8S277, D8S298 and D8S1771 was 32.6% (15/46), 44.7% (21/47), and 46.5% (20/43), respectively. LOH in plasma DNA was more frequently detected in the patients with intrahepatic cancer metastasis than those without metastasis (62.5 percent vs. 26.1 percent, P < 0.05); however, no statistically significant correlations were observed between LOH at these loci and other clinicopathological features analyzed in this study.

CONCLUSIONSLOH at D8S298 in plasma DNA may be a potential predictive marker of intrahepatic metastatic recurrence after surgical resection of the HCC.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; blood ; genetics ; Chromosomes, Human, Pair 8 ; DNA ; blood ; Female ; Humans ; Liver Neoplasms ; genetics ; Loss of Heterozygosity ; Male ; Middle Aged

OBJECTIVEGenetic factors are thought to be involved in the development of vitiligo. The aim of this study is to explore the possible genetic model of vitiligo by analyzing the genetic characteristics of 815 patients from Zhejiang province.

METHODSData for 815 patients with vitiligo together with their first- and second-degree relatives were obtained using a standardized questionnaire. All these information was requested to confirm the answers about family history in order to reduce the possibility of 'recall' bias. The 815 probands would include 411 (50.43%) males and 404 (49.57%) females with a varied age from 2 months to 71 years old. Since the information on general prevalence of vitiligo in this area was absent, a control group was set up to facilitate the calculations of heritability degree. 468 persons of the control group were from non-vitiligo population with a sex ratio of 241(male): 227(female) with varied age of 4 months to 80 years old. Both gender and age were comparable between the vitiligo and the control population. The inheritance pattern estimation, heritability calculation and complex segregation analysis were performed with Penrose method, Falconer regression method and SAGE-REGTL program.

RESULTSIn 815 vitiligo probands, 128 had and 687 had not family histories, with a heritability rate of 15.7%. The vitiligo prevalence in proband's first degree relatives was 2.580%, higher than the prevalence of 0.618% in second degree relatives, and both of them were higher than general prevalence: 0.192%. By Penrose method, the rates on different catagories were as follows: sibling prevalence rates s = 0.080 18; population prevalence rate q = 0.001 92; s/q = 41.76. The ratio of s/q did not approach 1/2q (260.42) or 1/4q (130.21), but approached 1/square root of q(22.82), suggesting vitiligo was consistent with a mode of polygenic inheritance. Using Falconer's method, heritabilities of vitiligo in first-and second degree relatives of probands were 59.61% (95% confidence interval 65.37-53.84) and 55.20% (95% confidence interval 43.88-66.52), respectively. The weighted average of heritability in all relatives was 58.7% (95% confidence interval 53.56-63.83). The results of complex segregation analysis suggested that major gene model including the Mendelian dominant, recessive and additive hypotheses were not rejected (P > 0.05). Purely environmental model and no transmission model were rejected at a 0. 001 significance level. According to AIC, Mendelian dominant inheritance was the best-fitted hypothesis.

CONCLUSIONGenetic factors played an important role in the occurrence of vitiligo, and the genetic model of vitiligo could serve as the polygenetic or multifactorial inheritance with major gene trait.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Male ; Middle Aged ; Models, Genetic ; Vitiligo ; epidemiology ; genetics ; Young Adult

This study was purposed to investigate the effect of crocin on the proliferation in vitro and immune function of dendritic cells (DC) derived from the bone marrow of children with acute leukemia. The mononuclear cells were isolated from bone marrow of leukemia children by Ficoll-Hypaque. The experiment was divided into six groups: blank control group (A), crocin 1.25 mg/ml group (B), cytokines (rhGM-CSF 75 ng/ml+rhIL-4 75 ng/ml+rhTNF-α 50 ng/ml) group (C), cytokines+crocin 0.3125, 1.25 or 5.0 mg/ml groups (D, E, F). The numbers of DC were counted and the phenotypes of DC were determined by flow cytometry on the ninth day of culture. The DC of different groups were mixed with T cells just separated from peripheral blood of another children with acute lymphoblastic leukemia, and cultured with rhIL-2 200 U/ml for 5 d. The function of DC was detected by mixed lymphocyte reaction (MLR). The results indicated that the test groups and control group all obtained a certain amount of typical DC, but the DC numbers in test groups were all higher than those in control group (P < 0.01). Cultured for 9 days, the rates of CD1a(+), CD83(+), and HLA-DR(+) in group C, D, E, F were higher than group A (P < 0.01). There was no statistically significant difference between A and B groups (P > 0.05). MLR showed that with the increasing of DC, the stimulation index of T cells in group A and B was not rising (P > 0.05); the stimulated index of T cells in group C and E was significantly rising, there was statistically significant difference between them (P < 0.01). When the number of stimulated cells was the same, the stimulation index of T cell in group E was the highest (P < 0.01). It is concluded that the capability of DC proliferation promoted by crocin alone is lower than that of its combination with rhGM-CSF, rhIL-4 and rhTNF-α, but the crocin can synergically promote the maturity of DC cooperating with rhGM-CSF, rhIL-4 and rhTNF-α. The DC induced by crocin can particularly enhance the proliferation of T cells.
Bone Marrow Cells ; cytology ; drug effects ; Carotenoids ; pharmacology ; Cell Proliferation ; drug effects ; Child ; Dendritic Cells ; cytology ; drug effects ; Humans ; Leukemia ; pathology ; Lymphocyte Culture Test, Mixed ; T-Lymphocytes ; cytology ; Tumor Cells, Cultured

OBJECTIVETo study the newborn's Apgar score in 'one minute' and relevant factors.

METHODSOne year inpatient woman from a Maternal and Child Health Hospital of Anhui province were selected by cluster sampling method and newborn asphyxia situation was investigated using Apgar score and self-designed questionnaire.

RESULTSThe Apgar score in 'one minute' which marking 8 to 10, 4 to 7 and 0 to 3 were found in 1875 (73.78%), 426 (16.77%) and 240 infants (9.45%) respectively. The average Apgar score in 'one minute' and five minutes were (7.69 +/- 2.27) and (9.01 +/- 1.89) respectively, The Apgar score in 'one minute' was significantly correlated with that of five minutes (Pearson coefficient correlation r = 0.841, P = 0.00). Ordinal regression analysis revealed that parturient age (OR = 1.04), being farmer (OR = 2.22), parity (OR = 1.26), assistant vaginal delivery (OR = 4.93), caesarean section (OR = 1.95), pregnancy-induced hypertension syndrome (OR = 1.42), albuminuria in gestational period (OR = 1.44), newborn being male (OR = 1.23), low birth weight (OR = 2.94), inborn abnormality (OR = 12.12), premature birth (OR = 1.22) and complications of delivery (OR = 5.04) were risk factors while the number of years under study (OR = 0.91), prenatal check-up (OR = 0.48), body length of newborn infant (OR = 0.88) and single birth (OR = 0.57) were protective factors.

CONCLUSIONApgar score in 'one minute' of newborn infant was affected by several factors as stated above. Health care program in earlier period toward community parturient should be strengthened in order to discover and control high risk factors of duration of pregnancy in earlier period.

Apgar Score ; Asphyxia Neonatorum ; epidemiology ; Epidemiologic Studies ; Female ; Humans ; Infant, Newborn ; Male ; Pregnancy ; Risk Factors ; Time Factors