Immunohistochemical method, immuno-electron microscopic technique and radioimmunoassay (RIA) were used for the demonstration of ?-endorphin-like immunoreactivity (?-EP-IR) and of the circadian variation of the ?-EP-IR level in rat pituitary. ?-ER-IR was located in all cells of the pituitary intermediate lobe. A few ?-ER-IR-positive cells were scattered in the anterior lobe, and no positive immunoreactivity was found in the posterior lobe. The controls were negative. Under electron microscope, ?-ER-IR-positive dense particles were mostly located in the secretion granules, and a few of them were scattered freely in the cytoplasm of the intermediate lobe cells. Both pituitary ?-EP-IR concentration and the basal pain threshold in rat showed circadian variations in the experiment done at 4-hr intervals over a 24-hr period (P

The heterogeneity is one of the main points in study on the mast cells. In the present paper, the heterogeneity of skin connective mast cells (CTMC) in comparison with that of gastrointestinal mucosal mast cells (MMC) with both toluidine blue staining and immunostaining was studied. The results showed that the CTMC fixed with formaldehyde could be demonstrated with routine toluidine blue staining, whereas the MMC could be demonstrated only with lower pH and prolonged staining duration. In immunostaining almost all of the CTMC showed serotoninimmunoreactivity, and only 10% of the CTMC showed substance P-immunoreactivity; while approximate 35% of the MMC showed serotonin-immunoreacivity and no substance P-immunoreactivity was found. The results indicate that the mast cell heterogeneity may be found in toluidine blue staining as well as in immunostaining.

In the present study the effects of electroacupuncture (EA) on substance P (SP), somatostatin (SOM) and serotonin (5-HT) immunoreactivity (IR) of APUD cells and nerve fibers in the pylorus and in the skin of "Zusanli" acupo- int of rats were studied using immunohistochemical technique. Forty rats were divided into 20 pairs, one of each pair for EA (bilateral "Zusanli" acupuncture for 20 min), the other one for control. After EA the pain threshold was mark- edly increased(P

Objective To investigate the relationship between acupuncture analgesia and cellular immunity, the acupuncture effect on expression of c fos mRNA, ppENKmRNA,iNOSmRNA and iNOS activity in the mouse peritoneal macrophage were studied. Methods Twenty BALB/c mice were randomly divided into 2 groups:a,the acupuncture group treated with 5Hz electroacupuncture (EA); b, the control group treated with no EA. The pain threshold was detected by K + ionophoresis method before and after EA or restraining. The experiment was performed by using in situ hybridization, NADPH NBT histochemistry, RNA dot blot and protein dot blot techniques. All dot blot signals were scanned for statistical analysis. Results The hybridization signals and iNOS activity appeared as granules in bluish violet color, localized in the cytoplasm of the macrophage. All dot blot signals were increased in the acupuncture group, when compared with those in the control group( P

After acupuncture,histamine flares are often found on skin arround acupuncture points in most patients,histamine and other biologically active substances are always present in the local soft tissues.“Meridian phenomena”,such as urticarious skin reaction or red line along the meridian may be shown in a few patients.It is well known that mast cells release histamine and other active mediators.The purpose of this paper is to observe the distribution of mast cells in the connective tissue at points in albino rats and if there is any difference in the distribution pattern of mast cells between adult and new born rats. 52 albino rats,including adult,new born and developing ones were investigated. Points selected on the lower extremities were identified by decreased electric resistance and were marked with indian ink:the electric resistance of skin beside points was detected simultaneously as control.The mast cells in the spread preparations of subcu- taneous connective tissue(fascia)in albino rats were fixed in alcohol-formalin and stained with toluidin blue.Most specimens of fascia were fixed at first and spread on the slides later,while some specimens were spread before fixation.No essential difference between both kinds of specimens was found. In the adult rats,the mast cells in fasciae are distributed not uniformly,they are oval in shape and are usually arranged in rows along the cell length with polarity. Many rows of mast cells are packed to form an elongated zone.Several zones oriented in the direction of the longitudinal axis of lower extremities are found.The mast cells are crowded in the field of marked points along the zone.Besides,it is found that the mast cells along small blood vessels are slender and fusiform in shape. However,in new born rats,mast cells are distributed uniformly and most of them are round in shape.The mast cells are arranged in random way without polarity.In the arrangement of mast cells neither row nor zone is found,and there is no difference in distribution of them everywhere. In the developing rats aged one month or so,the morphology of mast cells is similar to that of the new born,except that they are more numerous at the points. In the adult rats,electric resistance at various points is 260 kilo-ohm,while that of control is more than 500 kilo-ohm.In contrast with the adult,the electric resistance on any part of skin(at point or not)of the new born is equal,more than 500 kilo- ohm,and the electric resistance of points is decreased with development of albino rats in which an apparent decrease may be demonstrated in the rats aged about one month.It seems to be that there are parallel changes with growth of individuals between the morphological character of mast cells in the fasciae and functional property detected by electric resistance at points. It has been considered that the morphology and amount of mast cells may indicate the activity of metabolism of connective tissue.It suggests thence that the metabolism of fasciae at points may be more active and the points may be formed with the developed function in individuals after birth in albino rats.

In order to improve the technique for histochemical demonstration of the different lymphocyte patterns by alpha-naphthyl acetate esterase (ANAE) activity from a single microscopic slide, 10 kinds of comparative experiments were performed. The results showed as follows. The peripheral blood mononuclear cells first fixed in 0.4% glutaldehyde and postfixed in 15% formol calcium gave the best effect on preserving the enzyme activity and on connterstaning of the leucocyte nuclei. During formol calcium fixation, the erythrocytes were dissolved from the diluted blood film with PBS more easily than from the blood smear without any dilution. The lymphocytes were distributed more uniformly and densely in the former specimen than in the latter. When incubated with 0.5 mg alpha-naphthyl acetate/ml medium in hexoazotized pararosaniline for 1(1/2)～2 hrs at 37℃, the distinct ANAE patterns of circulating lymphocytes were more readily identified.In 28 healthy adults the differential counts of the lymphocyte ANAE patterns from the diluted blood specimen was similar to those found from the specimen of isolated lymphocytes. The focal pattern was about 56%, the diffuse pattern about 25% and the negative pattern about 19%; while the isolated lymphocyte specimen was compared with the diluted blood film from the same individuals, the focal pattern was decreased insignificantly, the diffuse pattern increased and the negative pattern decreased significantly.

Objective To investigate the association between microalbuminuria (MA) and high-sensitive C-reactive protein in essential hypertensive subjects. Methods Urinary albumin excretion rate (UAER)and the concentrations of high-sensitive C-reactive protein(hs-CRP) were measured. One hundred and eight patients with hypertension were divided into 2 groups: normoalbuminuria and microalbuminuria group. concentrations of hs-CRP was compared between the two groups , and the association between MA and hs-CRP was evaluated.Results Patients with microalbuminuria were significantly characterized by higher systolic blood pressure (SBP) ,pulse pressure(PP) ,circulating fibrinogen and hs-CRP. Evaluation of bivariate correlation indicated that UAER was positively correlated with SBP , PP , body mass index (BMI) ,Serum creatinine (SCr), fibrinogen and hs-CRP. Multivariate regression analysis demonstrated PP, SCr and hs-CRP were independent risks associated with UAER.Conclusions Microalbuminuria is a sensitive marker in essential hypertensive subjects with early renal injury .hs-CRP is an independent risk associated with UAER ,which suggests that inflammation correlate with early renal injury in essential hypertension.

Objective To explore the effects of vitamin C and niacinamide on the growth and differentiation of human primary cultured keratinocytes.Methods Normal human foreskin was used in this study.The epidermis was separated enzymatically from the dermis by thermolysin,and keratinocytes were isolated from the epidermis by digestion with trypsin plus EDTA.The single keratinocytes were cultured with undedying NIH-3T3 cells as feeder cells in a complete medium supplied with 50 mg/L (vitamin C group),niacinamide of 400 μmol/L(niacinamide group)or vehicle(control group).Immunocytochemistry and immunodot blot were performed using monoclonal antibodies directed against C-myc,cyclin D1,filaggrin and involucrin.Results The colony number was highest in vitamin C group,followed by the control group and niacinamide group,and the colony morphology in vitamin C group was similar to that in the control group,but distinct from that in the niacinamide group.A significant increase was noticed in the expression of C-myc,cyclin D1,filaggrin and involucrin in vitamin C-treated keratinocytes compared with the control keratinocytes(all P＜0.05);however,in niacinamide-treated keratinocytes,the expression of filaggrin was significantly enhanced(P＜0.01),that of involucrin remained unchanged(P＞0.05),while that of C-myc was depressed(P＜0.05).Conclusions These results demonstrate that vitamin C has a favorable effect on both the growth and differentiation of human keratinocytes,while niacinamide seems to only promote the differentiation but attenuate the growth of human keratinocytes.

Objective To explore the effects of natural avocado oil and olive oil on the proliferation and differentiation of HaCaT cells. Methods MTT assay was performed to determine the optimal work concentration of avocado oil and olive oil. Cultured HaCaT cells were divided into 3 groups, i.e., avocado oil group treated with avocado oil of 3% (v/v), olive oil group treated with olive oil of 3% (v/v), and control group without any treatment. Immunocytochemistry and immuno-dot-blot method were used to detect the expressions of c-myc, mitogen-activated protein kinase ( MAPK ), nuclear factor ( NF)-κB, filaggrin, involucrin and keratin10 in HaCaT cells. Results As immunocytochemistry showed, the mean grey values (staining intensity) of c-myc,MAPK, and NF-κB in HaCaT cells were 131.4 ± 6.6,136.3 ± 4.5 and 134.3 ± 5.2 respectively in the avocado oil group, 121.1 ± 4.5, 107.9 ± 7.3 and 106.4 ± 5.4 respectively in the olive oil group, significantly higher than that in the control group (101.9 ± 8.9,91.4 ± 5.1 and 94.3 ± 7.0, respectively, all P＜ 0.05), and the avocado oil group was higher than the olive oil group in all the above parameters (all P ＜ 0.05). Increased expressions of filaggrin, involucrin and keratin 10 were observed in the avocado oil group and olive oil group compared with the control group (all P＜ 0.05), and in the olive oil group than in the avocado oil group (all P＜ 0.05).The mean grey values of these proteins obtained by immunocytochemisty were significantly correlated with those obtained by immuno-dot-blot method in avocado oil group (r = 0.94, P ＜ 0.01 ) and olive oil group (r=0.97, P ＜ 0.01 ). Conclusions Certain concentrations of avocado oil and olive oil can promote the proliferation and differentiation of HaCaT cells; avocado oil is more capable to accelerate their growth and proliferation, and olive oil to enhance their differentiation.

The met-enkephalin (MEK), leu-enkephalin (LEK) and ?-endorphin (?-EP) immunoreactivities were detected in the circulating lymphocytes of 64 guinea pigs by PAP immunohistochemical method under the conditions of electroacupuncture, naloxone or exogenous opioid like peptides (OLP). Subsequently the ?-naphthyl acetate esterase (ANAE)was demonstrated on partial immunostained lymphocyte samples. The results showed that the OLP immunoreactivity with different intensity was found in approximate 96％ of the lymphocytes. There was a positive correlation between the ANAE-focal pattern lymphocytes and MEK-IIRL (intense immunoreactivity lymphocytes)or ?-EP-IIRL; whereas there was no significant correlation between the ANAE-focal pattern lymphocytes and LEK-IIRL. The MEK-IIRL, LEK-IIRL and ?-EP-IIRL were all increased markedly in the guinea pigs which underwent electroacupuncture at "Zusanli" points. There was no significant difference in MEK-IIRL, LEK-IIRL and ?-EP-IIRL, when the acupuncture group, naloxone group and control group were compared with each other. However, the LEK-IIRL, MEK-IIRL and ?-EP-IIRL could be depressed by naloxone, when the lymphocytes were preincubated with 10~(-7)mol/L of MEK, LEK or ?-EP in vitro. Hence, the naloxone group had a significant difference in MEK-IIRL, LEK-IIRL or ?-EP-IIRL when compared with the acupuncture group. A high peak of the lymphocyte MEK-IR among the range of 10~(-10)—10~(-8)mol/L of MEK preincubation in the acupuncture group when compared with the control group. It suggests that electroacupuncture may enhance activity of the unbound MEK receptor.