Objective To investigate the location,fine structure of melanocytes in human fetal scalp hair follicles.Methods The scalp with hair follicles was obtained from a dead fetus of 6 months of age,and divided into two parts.One part was embedded in paraffin,tissue sections were prepared with a width of 7 μm and stained with NKI/beteb,monoclonal antibodies to HMB-45,tyrosinase and tyrosinase-related protein 1(TRP1),respectively.The other part with hair follicles was treated with collagenase type Ⅱ 0.1 g/L and trypsin,then,cell suspension was collected and cultured.After 14-day culture,follicle melanocyte cells (FMC)were separated from keratinocytes by differential trypsinization,and fibroblasts were removed with geneticin.Following three times of pure passage,FMC were seeded and fixed on mica for scanning electron microscopy(SEM)and atomic force microscope(AFM)scanning.Results Histopathological examination showed that NKI/beteb positive cells located at the outer root sheath of human hair follicles,and these cells stained negatively for HMB-45,tyrosinase and TRP1 antibodies.However,in the hair bulb,lots of cells expressed HMB-45,tyrosinase and TRP1 antigens.After fibroblasts and keratinocytes were removed,two kinds of melanocytes remained in the culture:one was small in number and showed abundant melanin,which was lost after subsequent passage;the othgr was large in number and had no melanin initially,but proliferated very rapidly.After three passages,almost all the melanocytes were positive for NKI/beteb.As SEM and AFM showed,most cultured melanocytes appeared fusiform with two(rarely three)dendrites,and the cell body was round or oval with a few melanosomes scattered in but no clear secondary branches on the dendrites.Conclusions The melanocytes in outer root sheath of hair follicles from the fetal scalp are presumed as melanocyte stem cells or their progenies.In vitro,these cells proliferate very rapidly during early phases,but the morphology and function of them still remain immature,which is unfavorable for melanosome transport.

OBJECTIVETo observe the effect differences on post-stroke dysphagia among acupoint injection combined with neural electrical stimulation, acupoint injection, neural electrical stimulation, and swallowing training respectively, so eligible intervention can be applied to this kind of disease.

METHODSOne hundred and eight-three patients of post-stroke dysphagia were randomized into a comprehensive treatment group (42 cases), an acupoint injection group (44 cases), a neural electrical stimulation group (49 cases) and a swallow training group (48 cases) and were treated with the comprehensive therapy of acupoint injection and neural electrical stimulation, acupoint injection, neural electrical stimulation and swallowing training separately. The treatments for 10 days made one session. There were 3 days at the interval among treatment sessions and 3 sessions were required totally. The cases in those treatment groups were blankly controlled with the other 47 patients of post-stroke dysphagia. All the patients received basic rehabilitation treatment. The modified water swallowing test was conducted to assess the efficacy before treatment, 10 days after treatment and 30 days after treatment in each group separately. The clinical efficacy, score of water swallowing test and improvement in water swallow test were compared among the groups.

RESULTSAfter 10-day treatment, the differences in efficacy and score of water swallow test were not significant in each group (all P > 0.05). After 30-day treatment, the effective rate (94.29%, 33/35) in the comprehensive treatment group was apparently better than 68.75% (22/32) in the acupoint injection group, 80.00% (32/40) in the neural electrical stimulation group, 67.50% (27/40) in the swallowing training group and 42.86% (12/28) in the blank group separately. The score in water swallow test in the comprehensive treatment group was lower than that in each of the other groups (1.37 ± 0.60 vs 2.03 ± 1.00, 1.90 ± 0.90, 2.20 ± 0.72, 2.71 ± 0.90, all P < 0.05). The differences in the effective rate and score in water swallow test were not significant among the acupoint injection group, neural electrical stimulation group and swallowing training group (all P > 0.05), which indicated that the improvement in swallowing function in the comprehensive treatment group was significantly superior to the other groups (all P < 0.05).

CONCLUSIONThe comprehensive therapy of acupoint injection and neural electrical stimulation achieves the much better efficacy on post-stroke dysphagia.

Acupuncture Points ; Adult ; Aged ; Combined Modality Therapy ; Deglutition ; Deglutition Disorders ; drug therapy ; etiology ; physiopathology ; therapy ; Electric Stimulation Therapy ; Female ; Humans ; Male ; Middle Aged ; Stroke ; complications ; Treatment Outcome ; Vitamin B 12 ; administration & dosage ; Young Adult

Objective:To analyze and compare the volatile components in vinegar-processed schisandrae sphenantherae fructus and vinegar-processed schisandrae chinesis fructus.Methods:The volatile components in vinegar-processed schisandrae sphenantherae fruc-tus and vinegar-processed schisandrae chinesis fructus were extracted by headspace solid phase-microextraction (HS-SPME) and quali-tatively analyzed by GC-MS.Results:Totally 20 kinds of constituents were identified from vinegar-processed schisandra sphenanthera fructus, which accounted for 99.55%of the total volatile components , and totally 21 kinds of constituents were identified from vinegar-processed schisandra sphenanthera fructus, which accounted for 99.90% of the total volatile components .Conclusion: The type and content of volatile components in vinegar-processed schisandrae sphenantherae fructus and vinegar-processed schisandrae chinesis fructus are quite different , and the study can provide scientific basis for the two traditional Chinese medicinal materials .

Objective To investigate the changes of urine conductivity in common kidney diseases and the relationship between urine conductivity and cystatin C.Methods A total of 723 patients in the First Affiliated Hospital of Dalian Medical University were selected in this study and divided into IgA nephropathy group(81 patients),diabetic nephropathy group(104 patients),ne-phritic syndrome group(130 patients),multiple myeloma group(89 patients),chronic glomerulonephritis group(1 1 5 patients),renal calculus group(101 patients)and chronic renal failure group(103 patients).Other 205 healthy persons were recruited into control group.The urine samples were analyzed by Sysmex automatic urinary sediment analyzer UF 1000i to determine the urine conductivi-ty,while the serum samples were analyzed by Hitachi automatic biochemical analyzer 7600 to determine the cystatin C.All the re-sults were statistically analyzed by SPSS1 9.0 software.Results The urine conductivity in kidney patients was significant lower than that of healthy people (P <0.05).There was a significant negative correlation between urine conductivity and the cystatin C in the kidney patients(P <0.05).Conclusion The urine conductivity might serve as an important index in evaluating the kidney func-tion and urine concentration.

Objective To observe the therapeutic effects and assess the feasibility of radiofrequency thermo- coagulation denervation(TRD)mediated by digital subtraction angiography(DSA)for patients with lumbar facet joint syndrome(LFJS).Methods Eighty LFJS patients were randomly divided into two groups:a denervated group(groupⅠ)and a control group (groupⅡ).The lumbar facet joints of the patients in groupⅠwere treated with TRD,while those in groupⅡhad their lumbar facet joint cavities injected with the mixture of 1 mg betamethasone and 1 ml lidocaine.All patients were assessed using the Visual Analog Scale(VAS)before treatment and 30 rain,1 d, 1 w,1 m and 6 m after.Schober values were measured at 1 w,1 m and 6 m after treatment.The therapeutic effects and side effects were assessed at 6 m after treatment.Results VAS scores in groupsⅠandⅡat 30 min,1 d,1 w and 1 m after treatment were remarkably lower than before treatment.In addition,those in groupⅠ6m after treatment were also significantly lower(P＜0.01).Compared with those in the groupⅡ,VAS scores in groupⅠwere signifi- cantly decreased at 1 m and 6 m after treatment(P＜0.05 and P＜0.01,respectively).The schober values in groupⅠat 1 m and 6m were obviously increased in comparison with those in groupⅡ,while the therapeutic effects in groupⅠat 6m after treatment were better than those in groupⅡ(P＜0.01).There were no side effects,such as injuries to the nerve roots or dysfunctional dermal sensation,in either group.Conclusion TRD,mediated by DSA,has ther- apeutic effects on LFJS.Such treatment is safe,convenient,and gives long-term pain relief.

Objective To investigate the clinical utility of anti-dsDNA antibody in the diagnosis of systemic lupus erythematosus (SLE).Methods Total and high affinity anti-dsDNA antibodies of 431 serum samples were measured by total and high affinity anti-dsDNA antibody EIA kits.Modified SLE disease activity index (M-SLEDAI) was scored for each SLE patients at the time of serum collection.The agreements between total and high affinity anti-dsDNA EIA tests and the correlation of anti-dsDNA antibody levels with disease activity and clinical manifestations were analyzed.Statistical analysis was performed using x2 test,Spearman correlation,Mann Whitney U test & Fisher exact,Student t test.Results ① The overall agreement between total and high affinity anti-dsDNA antibodies was 93.3％,and the total agreement of SLE patients was 89.6％.Twenty-two out of 23 (95.7％) SLE patients with positive total anti-dsDNA antibody but high affinity antidsDNA antibody were inactive with the M-SLEDAI score less than 4.Four patients with other autoimmune diseases had positive total anti-dsDNA antibody but negative on high affinity anti-dsDNA antibodies.② The total and high affinity anti-dsDNA antibody levels were significantly positively correlated with disease activity (M-SLEDAI) and negatively correlated with serum C3 and C4 levels (P＜0.01).③ The ratio of high affinity to total anti-dsDNA antibody was significantly higher in patients with M-SLEDAI ≥4 or with active kidney damage (P＜0.01).Conclusion Both total and high affinity anti-dsDNA antibodies are significantly correlated with SLE disease activity and kidney damage.However,the high affinity anti-dsDNA antibody may be more specific for active SLE and helpful in the differential diagnosis with other autoimmune diseases.High affinity anti-dsDNA antibody is more valuable in monitoring SLE disease activity status,especially on kidney damage,compared to total anti-dsDNA antibodies.

To investigate the expression of the subunit p65 of NF-kappaB and inhibitor kappa B alpha (IkappaBalpha) in mouse uterus during peri-implantation, thereby investigating whether transient activation of nuclear factor-kappaB (NF-kappaB) takes place during embryo implantation in mice. Immunohistochemical technique was used to examine the expression and localization of p65 in endometrium or deciduas, and Western blot analysis was employed to detect the levels of IkappaBalpha protein in mouse endometrium or deciduas. P65 protein was detected in stromal cells, epithelial cells of endometrium as well as in myometrium. Staining was predominately seen in the cytoplasm of the cells. Staining intensity for p65 was stronger in the epithelial compartment than the stromal compartment and myometrium. Staining intensity increased slightly during pregnancy, and it reached a high level on pregnancy day 5 and day 8. In contrast to p65, the level of IkappaBalpha protein was lowest on pregnancy day 5 in all groups. Our results suggested that NF-kappaB may regulate embryo implantation by its transient activation in mice.
Decidua/metabolism ; Embryo Implantation/*physiology ; Endometrium/metabolism ; I-kappa B Proteins/*biosynthesis ; NF-kappa B/*biosynthesis ; Time Factors ; Uterus/*metabolism ; Uterus/physiology

The activity of matrix metalloproteinases (MMPs) in the uterine flushing and endometrial tissue of normal adult women wearing FCu-IUD (fixed Cu-IUD) or FICu-IUD (indomethacin-releasing FCu-IUD) was observed by using zymography on SDS-PAGE containing gelatin. The results showed that the activity and kinds of MMPs in FCu-IUD group were increased significantly as compared with themselves before being inserted FCu-IUD. However, compared with the FCu-IUD group, the activity of some kinds of MMPs in the FICu-IUD group was decreased significantly. These data suggest that IUD can enhance the activity of MMPs in human endometrium, intermediated by prostaglandins, and MMPs may have relation to IUD-induced menorrhagia and indomethacin reduces IUD-induced menorrhagia by partly inhibiting MMPs synthesis.
Endometrium/*enzymology ; Indomethacin ; Intrauterine Devices, Copper/*adverse effects ; Intrauterine Devices, Medicated/*adverse effects ; Matrix Metalloproteinases/*metabolism ; Uterine Hemorrhage/etiology ; Uterine Hemorrhage/prevention & control

OBJECTIVETo explore changes of B and T lymphocyte attenuator (BTLA), superoxide dismutase (SOD), catalase (CAT), total antioxidant capacity (TAOC), reactive oxygen species (ROS), reactive nitrogen species (RNS), malondialdehyde (MDA) in ankylosing spondylitis (AS) patients, and the effect of Xinfeng Capsule (XFC) on them.

METHODSTotally 120 AS patients were assigned to two groups according to random digit table method, the XFC group (3 XFC pills each time, 3 times a day) and the SASP group (4 SASP tablets each time, twice a day), 60 in each group. All patients were treated for 3 months. Another 60 healthy subjects were recruited as a healthy control group. The expression frequency and activation levels of BTLA were detected using flow cytometry. Serum oxidative stress indices (such as SOD and CAT, TAOC, ROS, RNS, MDA) and contents of cytokines [tumor necrosis factor α (TNF-α), IL-1β, IL-4, and IL-10] were detected using enzyme-linked immunoassay (ELISA). Erythrocyte sedimentation rate (ESR) was detected using Westergren method. High-sensitivity C-reactive protein (Hs-CRP) was detected using HITACHI 7060 type automatic biochemical analyzer. Clinical efficacies of ASAS 20 and BASDAI50 were assessed using VAS. Correlation analysis between scoring for quality of life and BTLA expression frequency was performed.

RESULTS(1) Clinical efficacies of ASAS 20 and BASDAI50 were significantly better in the XFC group than in the SASP group (P < 0.01). (2) Compared with the healthy control group, BTLA expressions in the peripheral blood of AS patients decreased significantly (P <0. 05); SOD, CAT, and TAOC values significantly decreased (P < 0.01, P < 0.05); ROS, RNS, and MDA values significantly increased (P < 0.01, P < 0.05); TNF-α, IL-1β, ESR, and Hs-CRP values significantly increased (P < 0.01); IL-4 and IL-10 values decreased significantly (P < 0.01, P < 0.05). (3) Compared with pre-treatment in the same group, BTLA/CD19 + B, BTLA/CD24 + B, SOD, TAOC, IL-4, SF-36 [physical functioning (PF), social functioning (SF), role limitation due to physical problems (RP), role limitation due to emotional problems (RE), body pain (BP), mental health (MH), vitality (VT), general health (GH)] were significantly elevated; ROS, MDA, TNF-α, ESR, Hs- CRP, VAS, BASDAI and BASFI, BAS-G were significantly lower in the peripheral blood of the two groups after treatment (P < 0.01, P < 0.05). Better effect was shown in the XFC group in elevating BTLA/CD19+ B, BTLA/CD24 + B, SOD, TAOC, IL-10, BP, MH, VT, and SF; and lowering ROS, IL-1β, MDA, TNF-α, ESR, Hs-CRP, VAS, BASDAI, BASFI, and BAS-G (P < 0.01, P < 0.05). (4) Pearson correlation analysis showed, BTLA/CD19 + B expression of the peripheral blood was positively correlated with SOD, CAT, TAOC, IL-4, IL-10, GH, RP, BP, and SF (r = 0.431, 0.325, 0.318, 0.316, 0.348, 0.314, 0.358, 0.318, 0.326, respectively, P < 0.05, P < 0.01), while it was negative correlated with ROS, MDA, TNF-α, IL-1β, ESR, VAS, and BASDAI (r = -0.342, -0.368, -0.334, -0.354, -0.324, -0.372, -0.342, respectively, P < 0.05, P < 0.01). BTLA/CD24 B expression of the peripheral blood was positively correlated with SOD, TAOC, IL-4, IL-10, GH, RP, BP, SF, RE, MH, VT (r = 0.358, 0.352, 0.372, 0.436, 0.435, 0.326, 0.352, 0.345, 0.326, 0.343, 0.332, respectively, P < 0.05, P < 0.01), while it was negative correlated with ROS, RNS, MDA, ESR, Hs-CRP, VAS, BASDAI, and BASFI (r = -0.447, -0.336, -0.405, -0. 395, -0. 358, -0.436, -0.338, -0.425, respectively, P < 0.05, P < 0.01).

CONCLUSIONXFC could improve BTLA expression in the peripheral blood of AS patients, negatively regulate activation and proliferation of B cells, and reduce abnormal immune responses and oxidative stress injury, thereby effectively alleviating joint stiffness and pain.

B-Lymphocytes ; physiology ; Blood Sedimentation ; C-Reactive Protein ; metabolism ; Capsules ; Catalase ; metabolism ; Cytokines ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Flow Cytometry ; Humans ; Interleukin-10 ; metabolism ; Interleukin-1beta ; metabolism ; Interleukin-4 ; metabolism ; Malondialdehyde ; metabolism ; Oxidative Stress ; Quality of Life ; Reactive Oxygen Species ; Spondylitis, Ankylosing ; drug therapy ; Superoxide Dismutase ; metabolism ; T-Lymphocytes ; physiology ; Tumor Necrosis Factor-alpha ; metabolism

Objective To study the changes in the immune function of advanced hepatic carcinoma patients after thermochemoembo-lization via hepatic artery. Methods Forty advanced hepatic carcinoma patients were randomized divided into 2 groups. A group (n=20) was treated by perfusion with ADM(40mg) + MMC(10mg) in 37℃ 9% sodium chloride solution via hepatic artery, and B group (n=20) was treated by perfusion with ADM(40mg) + MMC(10mg) in 60℃ 9% sodium chloride solution via hepatic artery, then all were embolized with some quantity of lipiodol. Flow cytometry (FCM) was used to detect the changes in percentages of T-lymphocyte subsets (CD3<'3, CD4<'+, CD8<'+) and NK cells, and the contents of soluble interleukin-2 receptor (sIL-2R) were detected by double sandwich ELISA before and after therapy. LDH enzyme-release assay was used to detect cytotoxic activity of NK cells. Results Compared with pre-therapy group or A group after therapy, the percentage of CD4<'+ cells of B group after therapy significantly increased (P<0.05), the percentages of CD8<'+ cells and the content of sIL-2R of B group after therapy markedly decreased (P<0.05). The percentages of NK cells and cytotoxic activity NK cells of B group after therapy were significantly higher than those from pre-therapy group or A group (P<0.05). Conclusion Intra-arterial thermochemoembolization may activate and improve cell-mediated immune function with a certain degree.