[Debridement combined with negative-pressure wound therapy and local flap for treating a case of stingray sting].

On January 10, 2018, a 35-year-old male patient with a 3 day stingray stinger injury in his left thigh was admitted to Xuzhou Central Hospital. At the time of admission, the patient's left thigh was red, swollen, and painful. On the day of admission, the patient underwent emergency operation in the outpatient operating room for local debridement to remove the infected and necrotic tissue. After the routine dressing change and the wound got better, surgical debridement and negative-pressure wound therapy were performed, and finally local flap was used to repair the wound. On the 14th day after the flap repair operation, the suture was removed and the patient was discharged. After half a year follow-up, the appearance of the operation area recovered well. This case suggests that the timely and correct pre-hospital treatment, thorough and timely debridement, and systemic antibiotic application are important means to reduce further injury of toxin. The local flap can achieve satisfactory result on wound healing after the wound bed is prepared by negative-pressure wound therapy.

Knockdown of microRNA-103a-3p inhibits the malignancy of thyroid cancer cells through Hippo signaling pathway by upregulating LATS1.

MicroRNA (miR)-103a-3p has been shown to be involved in the development and progression of several types of cancer. However, the role of miR-103a-3p in thyroid cancer remains unclear. This study investigated the effects of miR-103a-3p on the biological characteristics of thyroid cancer cells and related mechanisms. In the present study, we found that the expression of miR-103a-3p was increased in thyroid cancer tissues compared to that in non-cancerous tissues. Additionally, the expression of miR-103a-3p in thyroid cancer cell lines (TPC-1, SW579, BHT101, K1) was markedly higher than that in the human thyroid cell line (Nthy-ori3-1). Silencing of miR-103a-3p obviously inhibited proliferation, migration, and invasion and promoted apoptosis of BHT101 cells. miR-103a-3p upregulation promoted the proliferation, migration, and invasion and inhibited apoptosis of K1 cells. Mechanistically, LATS1 was identified as a functional target of miR-103a-3p, and miR-103a-3p negatively regulated LATS1 expression. miR-103a-3p knockdown (or upregulation) partially reversed the effects of LATS1 knockdown (or overexpression) on proliferation, apoptosis, migration, and invasion of thyroid cancer cells. LATS1 knockdown inhibited the phosphorylation of YAP in BHT101 cells and promoted the nuclear translocation of YAP. Whereas, miR-103a-3p downregulation reversed the inhibitory effect of LATS1 knockdown on the Hippo signaling pathway. Moreover, overexpression of LATS1 induced YAP phosphorylation in K1 cells and inhibits nuclear translocation of YAP, and the upregulation of miR-103a-3p reversed this effect. The knockdown of miR-103a-3p inhibited tumor growth and progression in vivo. Taken together, knockdown of miR-103a-3p inhibits proliferation, migration, and invasion and promotes apoptosis of thyroid cancer cells through the Hippo signaling pathway by upregulating LATS1.

Carfilzomib combined with ex vivo-expanded patient autologous natural killer cells for myeloma immunotherapy.

Natural killer (NK) cell-based immunotherapy is promising, as NK cells are in the first line of defense against cancer and capital of lysing tumor cells without pre-stimulation. However, NK cells from multiple myeloma (MM) patients are always deficient in numbers and the expression of certain activating receptors, disabling them in cytotoxicity against the cancer. Therefore, effective strategies to expand NK cells and increase NK cell-mediated cytotoxicity against MM are significant. Here, NK cells were efficiently expanded from peripheral blood mononuclear cells (PBMCs) of newly diagnosed MM patients after co-culture with irradiated K562 cells transfected with 41BBL and membrane-bound interleukin (IL)-15 (K562-mb15-41BBL) in the presence of 200 IU/ml human IL-2. The ex vivo-expanded NK cells were demonstrated to vigorously kill both MM cells and autologous primary MM cells without significant lysis of patient normal cells. Further exploration revealed a significant increase in cell surface expression of most activating receptors of NK cells and indicated that expanded NK (exp-NK) cell killing of MM cells was mediated by perforin/granzyme. NK cells are capital of lysing human leukocyte antigen (HLA) I-deficient tumor cells and carfizomib, a selective proteasome inhibitor approved for the treatment of relapsed/refractory MM patient, down-regulates the expression of HLA class I, thus enhancing NK cell-mediated lysis in MM. Here, we found for the first time that carfizomib dramatically augmented ex vivo exp-NK cell cytotoxicity against patient autologous MM cells, suggesting the use of exp-NK alone or in combination with the drug to treat MM patient.

[Effect analysis of anterior cervical operation for severe cervical kyphosis].

Objective: To determine the feasibility and safety of anterior cervical decompression and fusion in severe cervical kyphosis treatment. Methods: Totally 29 patients with severe cervical kyphosis(Cobb angle>50°) underwent anterior cervical decompression and fusion from June 2008 to May 2016 were studied retrospectively. There were 19 males and 10 females. The average age was 32.6 years ranging from 14 to 53 years. According to the etiology, 12 patients had iatrogenic deformity (11 had post-laminectomy cervical kyphosis, 1 had kyphosis due to anterior graft subsidence), 5 had neurofibromatosis, 4 had infective kyphosis, 8 had idiopathic cervical kyphosis. The curvature of cervical angle was measured by two-line Cobb method. The severity of cervical kyphosis was evaluated by kyphosis index (KI). Parameters including kyphosis levels, the apex of the kyphosis, C(2-7) sagittal vertical axis(SVA) and T(1) slope were also measured on lateral radiographs in the neutral position in each patient. The pre- and post-operative Japanese Orthopaedic Association(JOA) scores, visual analogue scale (VAS) of neek pain, neck disability index (NDI) and cervical alignment were compared. All patients were treated by skull traction. Motor evoked potential and somatosensory evoked potential were applied intraoperation as the spinal cord monitor. Results: Skull traction was performed for an average of 6.3 days. The mean vertebral number in kyphotic region was 4.7. The average operation time was 155 minutes and blood loss was 135 ml. The preoperative C(2-7)Cobb angle was 46.6°±18.1° in average. It was reduced to 11.4°±6.4° in average after operation. The Cobb angle of operation region was 72.9°±19.6° in average before operation. It was reduced to 11.2°±6.4° in average after operation. The kyphosis region correction rate was 84.6%. The mean preoperative C(2-7)SVA changed from (3.8±14.6) mm to (12.6±7.8) mm postoperatively. The mean preoperative T(1) slope changed from -10.6°±16.4° to 7.1°±14.9° postoperatively. The average postoperative C(2-7) Cobb angle, Cobb angle of kyphosis region, KI, C(2-7) SVA and T(1) slope changed significantly compared with preoperation (F=12.700-218.200, all P<0.01). The average postoperative JOA, VAS and NDI scores improved significantly compared with preoperation (F=225.500, 217.900, 131.200, all P<0.01). Conclusion: For severe cervical kyphosis, anterior correction is a safe and effective technique, sufficient decompression will be achieved.

[Effect of specific immunotherapy on the psychological health level and quality of life in patients with allergic rhinitis].

Objective: To study the effect of specific immunotherapy on the psychological health level and quality of life in patients with allergic rhinitis(AR).Method:Selected 97 cases diagnosed as moderate to severe persistent AR patients, were treated with specific immunotherapy for one year. All patients received the evaluation with symptom check list 90(SCL-90) and rhinoconjunctivitis quality of life questionnaire(RQLQ) before specific immunotherapy, six, and 12 months after specific immunotherapy.Result:The total scores, scores of somatization, obsessive, anxiety, depression and phobia in SCL-90 of AR patients after 12 months treatment were significantly lower than that before treatment（P < 0.05）. Total score of quality of life and subitem score in RQLQ of AR patients after 12 months treatment were obviously lower than that before treatment (P < 0.05）.Conclusion:Specific immunotherapy can effectively alleviate the clinical symptoms and improve psychological health level and quality of life of AR patients.

[The dynamic change of cytokines associated with the speccificimmunotherapy and its clinical significance].

Objective:This project aimed to study the dynamic change of the cytokines associated with specific immunotherapy(SIT) pre- and post-SIT.Searching for immune regulatory indicators would used in SIT.Method:One hundred cases who had accepted SIT were enrolled in the project.Data of serum specific IgE and cytokines were statistically analyzed.In the three periods,pre-SIT,17 weeks post-SIT,57 weeks post-SIT,the levels of the eight kinds of cytokines(IL-4,IL-5,IL-8,IL-10,IL-13,IL-17,IFN-γ and TNF-α)were detected,and the dynamic change of the nasal symptoms score were analyzed.Result:The six kinds of cytokines(IL-5,IL-8,IL-10,IL-13,IL-17 and TNF-α)had no significant difference before and after SIT.The level of house dust mite sIgE level was positively correlated with serum IL-5 when the SIT pre-treatment and 57weeks (P<0.05).Pre-treatment and in 17 weeks after treatment,serum IL-5,IL-17 content difference and reduce the magnitude of nasal symptom scores were positively correlated (P<0.01).In 17 weeks of treatment and 57 weeks of treatment,difference of serum IL-10,IL-13,TNF-α levelsand the difference of nasal symptom scores were negatively correlated(P<0.01).Pre- treatment and 57 weeks,difference of serum IL-13,IL-17,TNF-α and the difference of nasal symptom scores were positively correlated (P<0.05),serum IL-10 levels of difference between the nose ministry of magnitude lower symptom scores were negatively correlated (P<0.01).Conclusion:The cytokines (IL-4,IL-5,IL-8,IL-10,IL-13,IL-17,IFN-γ and TNF-α) associated with the SIT play an important role in allergy and can objectively reflect the immune status during SIT.

Overexpression of clusterin correlates with tumor progression, metastasis in gastric cancer: a study on tissue microarrays.

Clusterin (CLU) is expressed in a wide variety of human tissues and fluids. Overexpression of cytoplasmic clusterin (sCLU) has been implicated in cancer development and progression. The aim of the present study is to evaluate the association of sCLU overexpression with clinicopathological features of human gastric carcinomas (GC).We constructed a gastric cancer tissue microarray containing 173 primary gastric carcinomas and 70 paired non-neoplastic mucosa specimens. The expression of sCLU was studied by immunohistochemistry. The correlations between sCLU expression and clinicopathological features, p53 abnormality, as well as Ki67 activation were analyzed. Overexpressions of sCLU was detected in 28.5% (n=165) of primary GCs by immunohistochemical staining, but not in non-neoplastic mucosa. Clinical association study found that overexpression of sCLU was significantly correlated with lymph-node metastasis (p < 0.001), tumor invasion (p < 0.001) and TNM stage (p < 0.001). In Kaplan-Meier survival analysis, overexpression of sCLU was significantly correlated with unfavorable survival in advanced GCs (p < 0.03). Furthermore, the association of sCLU with abnormal expression of p53 was ascertained. These results suggested that overexpression of sCLU was involved in the progression of GC and it's oncogenic function might be associated with p53 abnormality. Overexpression of sCLU seems to be related with patient's shorter survival in late stage GC.

Specific inhibition of kynurenate synthesis enhances extracellular dopamine levels in the rodent striatum.

Fluctuations in the endogenous levels of kynurenic acid (KYNA), a potent alpha7 nicotinic and NMDA receptor antagonist, affect extracellular dopamine (DA) concentrations in the rat brain. Moreover, reductions in KYNA levels increase the vulnerability of striatal neurons to NMDA receptor-mediated excitotoxic insults. We now assessed the role of a key KYNA-synthesizing enzyme, kynurenine aminotransferase II (KAT II), in these processes in the rodent striatum, using KAT II KO mice-which have reduced KYNA levels-and the selective KAT II inhibitor (S)-4-(ethylsulfonyl)benzoylalanine (S-ESBA) as tools. S-ESBA (applied by reverse dialysis) raised extracellular DA levels in the striatum of KYNA-deficient mice threefold and caused a much larger, 15-fold increase in wild-type mice. In the rat striatum, S-ESBA produced a 35% reduction in extracellular KYNA, which was accompanied by a 270% increase in extracellular DA. The latter effect was abolished by co-infusion of 100 nM KYNA. Intrastriatal S-ESBA pre-treatment augmented the size of a striatal quinolinate lesion by 370%, and this potentiation was prevented by co-infusion of KYNA. In separate animals, acute inhibition of KAT II reduced the de novo synthesis of KYNA during an early excitotoxic insult without enhancing the formation of the related neurotoxic metabolites 3-hydroxykynurenine and quinolinate. Taken together, these results provide further support for the concept that KAT II is a critical determinant of functionally relevant KYNA fluctuations in the rodent striatum.

Competitive interaction between two functional S-haplotypes confer self-compatibility on tetraploid Chinese cherry (Prunus pseudocerasus Lindl. CV. Nanjing Chuisi).

Self-incompatibility (SI) has been studied extensively at the molecular level in Solanaceae, Rosaceae and Scrophulariaceae, all of which exhibit gametophytic self-incompatibility (GSI). In the present study, four PpsS-haplotypes (Prunus pseudocerasus S-haplotypes) comprising at least two genes, i.e., PpsS-RNase (P. pseudocerasus S-RNase) and PpsSFB (P. pseudocerasus S-haplotype-specific F-box) have been successfully isolated in tetraploid P. pseudocerasus Lindl. CV. Nanjing Chuisi ("NC") which exhibited self-compatibility (SC), and its S-genotype was determined as S-1/S-3'/S-5/S-7. These PpsS-RNases, which were expressed exclusively in style, shared the typical structural features with S-RNases from other Prunus species exhibiting GSI. All PpsSFBs showed similar structure characteristics of SFBs from other Prunus species, and matched with the necessary conditions for pollen S-determinant. No mutations leading to dysfunction of S-haplotype were found in their full-length c-DNA sequences, except for PpsS-3'-haplotype which was not amplified by PCR. These four S-haplotypes complied with tetrasomic inheritance. Diploid pollen grains with S-genotypes S-7/S-1, S-7/S-5 and S-1/S-5 can grow the full length of the style after self-pollination, while pollen grains with S-3'/S-7, S-3'/S-1 and S-3'/S-5 cannot. These results suggest that PpsS-haplotypes-1, -5 and -7 are functional, and that competitive interaction between two of them confer self-compatibility on cultivar "NC". Furthermore, in terms of recognition specificity, diploid pollen grains carrying PpsS-3'-haplotype are equal to monoploid pollen grains carrying the other functional S-haplotype.

Focal microinjection of gamma-acetylenic GABA into the rat entorhinal cortex: behavioral and electroencephalographic abnormalities and preferential neuron loss in layer III.

Neuron loss in layer III of the entorhinal cortex (EC) occurs in patients with temporal lobe epilepsy and in several animal models of the disease and may play a role in the development of spontaneously recurring seizures. This damage can be reproduced in rats by a focal microinjection of the indirect excitotoxin aminooxyacetic acid into the EC (Neurosci. Lett., 147: 185, 1992). We have now examined a similar but approximately 20 times more potent toxin, gamma-acetylenic GABA (GAG), for its ability to produce seizures and neurodegeneration in the rat EC. EEG activity was recorded continuously for 48 h after a focal injection of 4 micrograms GAG into the rat EC. Seizure episodes, spiking, and other irregularities occurred with a latency of 150 min. Behavioral abnormalities were observed in all animals and were always accompanied by EEG seizures. The behavioral changes subsided gradually, but EEG seizures continued up to 24 h after GAG treatment. Nissl and silver-stained tissue sections obtained 2-3 days after the injection of 4 micrograms GAG revealed neuron loss which preferentially affected the medial part of layer III of the EC, and caused a modest lesion in the hilar region of the ventral hippocampus. The neurodegenerative potency of GAG, in contrast to the effects of aminooxyacetic acid, was not influenced by the depth of anesthesia during surgery. A slight increase in the dose of GAG (to 5 micrograms) resulted in more severe behavioral seizures, causing generalized convulsions with salivation and loss of righting posture in 3 of 13 rats. These animals also showed a marked enlargement of the lesioned area, with substantial neuronal loss occurring in layer III of the EC, in the hilus of the dentate gyrus, and occasionally also in homotopic structures of the contralateral hemisphere. Seizure activity and lesions induced by 4 micrograms GAG were prevented by the NMDA receptor antagonist Dizolcipine (MK-801) (4 mg/kg, i.p., 10 min before and 12 h after GAG). These data support the notion of a close correlation between the occurrence of seizures and neuronal loss in layer III of the EC. Taken together, the study suggests that intraentorhinal injections of GAG may provide an advantageous model for the study of epileptogenic and epileptic mechanisms.

[Gene expression and distribution in mouse abdominal cavity mediated by adenovirus].

Infection and expression of recombinant human adenovirus in mouse abdominal cavity was reported. After adenovirus vector Ad/RSV-beta-gal harboring the E. coli lacZ marker gene was injected into mice abdominal cavity, the peritoneal surface of jejunum, ileum, colon, uterus, liver, spleen, stomach, bladder, abdominal wall, diaphragm and testis was found large patches of lacZ-positive cells. But the adenovirus vector was not able to penetrate the peritoneum, as demonstrated by histochemical staining. Another adenovirus vector Ad/RSV-tk harboring the HSV-tk gene was injected into mouse abdominal cavity and the mouse was treated with ACV. No acute toxic reaction was observed. Based on these data, the feasibility of gene therapy of malignant tumor within abdominal cavity with adenovirus mediated TK/GCV system was discussed.

Alpha-Chain cross-linking in fibrin(ogen) Marburg.

The fibrinogen structural variant, Marburg (A alpha 1-460B beta gamma)2, is comprised of normal B beta and gamma chains but contains severely truncated A alpha chains that are missing approximately one half of their factor XIIIa cross-linking domain. Immunochemical studies of fibrin(ogen) Marburg were conducted to characterize the degree to which deletion of a defined A alpha-chain segment, A alpha 461-610, can affect the process of fibrin stabilization, ie, the factor XIIIa-mediated covalent interaction that occurs between alpha chains of neighboring fibrin molecules and between alpha chains and alpha 2 antiplasmin (alpha 2PI). The ability of Marburg (and control) alpha chains to serve as a substrate for factor XIIIa and undergo cross-linking was examined in an in vitro plasma clotting system. The capacity for alpha-chain cross-linking was evaluated both as the covalent incorporation of the small synthetic peptide, NQEQVSPLTLLK (which represents the first 12 amino acids of alpha 2PI and includes the factor XIIIa-sensitive glutamine residue responsible for the cross-linking of alpha 2PI to fibrin), and as the appearance of native (ie, natural), high-molecular-weight, cross-linked alpha-chain species. Antibodies specific for the (A)alpha and gamma/gamma-gamma chains of fibrin(ogen) and for the peptide and its parent protein, alpha 2PI (68 kD), were used as immunoblotting probes to visualize the various cross-linked products formed during in vitro clotting. Recalcification of Marburg plasma in the presence of increasing concentrations of peptide resulted in the formation of peptide-decorated Marburg alpha-chain monomers. Their size at the highest peptide concentration examined indicated the incorporation of a maximum of 3 to 4 mol of peptide per mole of alpha-chain. In the absence of alpha 2PI 1-12 peptide, the alpha chains of Marburg fibrin cross-linked to form oligomers and polymers, as well as heterodimers that included alpha 2PI. Both the peptide-decorated monomers and the native cross-linked alpha-chain species of Marburg fibrin were smaller than their control plasma counterparts, consistent with the truncated structure of the parent Marburg A alpha chain. Collectively, the findings indicate that, although deletion of the A alpha chain region no. 461-610 in fibrinogen Marburg prevents formation of an extensive alpha polymer network (presumably due to the absence of critical COOH-terminal lysine residues), it does not interfere with initial events in the fibrin stabilization process, namely, factor XIII binding and the ability of alpha chains to undergo limited cross-linking to one another and to alpha 2PI.

Induction of microsomal and peroxisomal enzymes by dehydroepiandrosterone and its reduced metabolite in rats.

Dehydroepiandrosterone (DHEA) given to rodents in pharmacological doses induces several hepatic enzymes including cytochromes P4504A, NADPH:P450 oxidoreductase, palmitoyl coenzyme A oxidase, and other enzymes associated with the peroxisomal beta-oxidation pathway, leading to peroxisome proliferation and development of hepatocellular carcinoma in rodents. Comparison of the inductive potency of DHEA and other intermediates of the steroid biosynthetic path demonstrated that only DHEA, 5-ene-androstene-3 beta,17 beta-diol (ADIOL), and to a lesser extent, 17 alpha-hydroxypregnenolone, a precursor of DHEA, induce cytochromes P4504A protein and other enzymes associated with the peroxisome proliferative response in vivo. ADIOL exerted its inductive response at a somewhat lower dosage than DHEA, whereas ADIOL and DHEA both induced the microsomal enzymes (P4504A and its oxidoreductase) at somewhat lower dosages than those required to induce peroxisomal enzymes. Northern analysis demonstrated increases in the mRNAs encoding the cytochromes P4504A (> 20-fold) and NADPH:P450 oxidoreductase (> 10-fold) in the livers of DHEA- and ADIOL-treated rats. Run-on transcription analysis demonstrated that DHEA induces CYP4A gene expression 11-fold at the level of transcription initiation. Comparison of the responsiveness of individual rat CYP4A genes (4A1, 4A2, and 4A3) to DHEA and ADIOL in immature versus mature male rats revealed 2-3-fold higher levels of induced CYP4A1 and 4A3 mRNAs in immature rat livers. In contrast, hepatic CYP4A2 mRNA was induced to 6-10-fold higher levels in mature rats. No basal or significant inducible expression of mRNA for CYP4A1 and 4A3 was noted in rat kidney. Significant basal levels of kidney CYP4A2 mRNA were observed only in mature animals, where they were inducible by ADIOL and DHEA to a 3-5-fold greater extent than in the kidneys of immature rats. These studies demonstrate developmental differences in the responsiveness of CYP4A mRNA levels to DHEA and ADIOL in rat kidney and liver. Moreover, the striking inducibility of CYP4A protein and mRNAs, together with the increased rates of synthesis of nascent CYP4A mRNA transcripts in hepatic nuclei from DHEA-treated rats, establish that DHEA increases the expression of these microsomal enzymes at the transcriptional level.

gamma-Acetylenic GABA produces axon-sparing neurodegeneration after focal injection into the rat hippocampus.

In exploring the recently discovered phenomenon of indirect excitotoxicity, we noted that intrahippocampal injections of the nonspecific aminotransferase inhibitor gamma-acetylenic GABA (GAG; 60-240 nmol) caused excitotoxic lesions in rats. When assessed 3 days following the injection, GAG was shown to be approximately equally toxic to CA3/hilar neurons and CA1 pyramids, while CA2 neurons and granule cells were clearly less vulnerable. Choline acetyltransferase activity, a marker of extrinsic afferents, remained unchanged in the GAG-lesioned hippocampus, indicating the axon-sparing nature of the insult. In contrast, a lesion caused by 240 nmol of GAG resulted in a significant reduction in 3H-MK-801 binding, which was used as a marker for NMDA receptor-bearing hippocampal neurons. GAG-induced lesions were blocked by the NMDA receptor antagonists MK-801 and AP7 but were not influenced by the nature of the anesthetic used during surgery. Iontophoretic application of GAG did not excite CA1/CA3 cells in the rat hippocampus. In vitro, GAG proved to be a relatively potent inhibitor (IC50: 43 microM) of kynurenine aminotransferase, the biosynthetic enzyme of the endogenous neuroprotectant kynurenic acid. GAG also inhibited the neosynthesis of kynurenic acid in hippocampal slices (IC50: 790 microM). Thus, GAG shares several characteristics of the recently described indirect excitotoxin aminooxyacetic acid (AOAA; Exp. Neurol. 113: 378, 1991). GAG and AOAA appear to belong to a new family of excitotoxic agents which produce lesions indirectly by metabolic derangement and/or inhibition of kynurenate production.

Induction of microsomal NADPH-cytochrome P-450 reductase and cytochrome P-450IVA1 (P-450LA omega) by dehydroepiandrosterone in rats: a possible peroxisomal proliferator.

Dehydroepiandrosterone (DHEA) is a naturally occurring C19-steroid that is found in the peripheral circulation of mammals, including humans. The feeding of DHEA to rodents has been shown to inhibit chemical carcinogenesis in colon, liver, and lung. Therefore, the effect of DHEA on hepatic enzyme activities that are associated with carcinogen metabolism was assessed. Microsomal NADPH-cytochrome P-450 reductase activity and the content of cytochrome b5 were induced 1.8- and 1.4-fold, respectively, upon feeding male Sprague-Dawley rats a synthetic diet containing 0.45% DHEA (w/w). No significant changes in total content of microsomal cytochrome P-450 or the activities of microsomal NADH-cytochrome b5 reductase and cytosolic or microsomal NAD(P)H-quinone oxidoreductase were noted at day 7 of feeding. Cytosolic glutathione S-transferase activity was decreased to 68% of control activity. Administration of DHEA p.o. or by i.p. injection for 5 days led to the same extent of induction of NADPH-cytochrome P-450 reductase activity. Maximal induction of this flavoprotein reductase was noted between days 3 and 4 of feeding or at a dose of 80-120 mg/kg i.p. A small but statistically significant increase in total microsomal cytochrome P-450 was observed after DHEA administration i.p. Rats fed DHEA had a slower growth rate compared with rats fed control diet, whereas rats treated with DHEA i.p. had growth rates identical to those of controls. The liver weights of rats given DHEA by p.o. or i.p. routes were increased significantly compared to those of control rats. Pair feeding of rats with DHA-containing or control diets served to demonstrate that the levels of induction of hepatic microsomal NADPH-cytochrome P-450 reductase and at least one form of cytochrome P450 (P-450IVA1) were the same as those seen in livers of rats fed DHEA ad libitum. This finding suggested that the induction of the flavoprotein and at least one form of the cytochrome was not due to caloric restriction. The increase in NADPH-cytochrome P-450 reductase content of liver microsomes prepared from rats either fed or treated i.p. with DHEA was also observed by Western blotting techniques. DHEA did not appear to induce any of the major forms of rat liver microsomal cytochrome P-450 that are normally increased by either phenobarbital, beta-naphthoflavone, or dexamethasone pretreatment of rats in vivo. However, the measurement of androstenedione and testosterone metabolism in vitro showed pronounced decreases in the 16 alpha-hydroxylase activities of liver microsomes following DHEA feeding.(ABSTRACT TRUNCATED AT 400 WORDS)

[Epithelioid sarcoma--immunohistochemical and ultrastructural study].

Five cases of epithelioid sarcoma are reported, of which four were studied by immunohistochemistry and one by electron microscopy. Immunohistochemically, the results of cytokeratin showed that these four cases were positive to different degrees by polyclonal cytokeratin and three out of four revealed positive results with low molecular weight cytokeratin monoclonal antibody. Two cases were positive with carcinoembryonic antigen and the other two were negative. The immunohistochemical stain was helpful for differential diagnosis from other sarcomas, such as malignant fibrous histiocytoma and rhabdomyosarcoma. By electron microscopy, tumor cells could be divided into light and dark cells which possessed more or less cell organelles, intermediate microfilaments in the cytoplasm and numerous pinocytic vesicles lying along the cell membrane. The occasional presence of poorly developed desmosomes or the absence of tonofibril bundles and glandular structures can rule out the diagnosis of carcinoma. One of these cases was originally diagnosed as metastatic carcinoma to the skin by light microscopy, which was refuted by electron microscopy, and the keratin positive reaction by immunohistochemistry was helpful for the diagnosis of epithelioid sarcoma. Therefore, the authors suggest that immunohistochemical and electron microscopic studies be used for diagnosing difficult cases of epithelioid sarcoma. From our observation, we consider that the cells of epithelioid sarcoma may derive from undifferentiated mesenchymal cells possessing the potentials of differentiating into synovioblasts, histiocytes or fibroblasts.

ATP as a marker of excitotoxin-induced nerve cell death in vivo.

In an attempt to find an marker for nerve cell death in vivo, the ATP content was measured in the rat dorsal hippocampus within hours or days following in the local injection of the excitotoxins quinolinic or kainic acid. Beginning or completed neuronal degeneration is accompanied by significant decreases in ATP levels. Selective blockade of the quinolinic acid-induced decrement in ATP content by D-(-) 2-amino-7-phosphonoheptanoic acid indicates that ATP measurements may of value for the rapid in vivo screening of the anti-neurotoxic properties of pharmacologically distinct excitatory amino acid receptor antagonists.