Concurrence of novel mutations causing Gilbert's and Dubin-Johnson syndrome with poor clinical outcomes in a Han Chinese family.

Dual-hereditary jaundice (Dubin-Johnson syndrome (DJS) and Gilbert's syndrome (GS)) is a rare clinical entity resulting from defects of the ATP binding cassette subfamily C member 2 (ABCC2) and UDP glucuronosyltransferase family 1 member A1 (UGT1A1) genes with autosomal recessive inheritance. In this study, we aimed to investigate the mutation profiles and characterize the phenotypes in a Han Chinese family with DJS and GS. Genetic screening for variants in the ABCC2 and UGT1A1, immunohistochemistry for expression of ABCC2, and histopathological examination were carried out. The proband and his brother had unconjugated and conjugated hyperbilirubinemia after birth. The proband's sister had only conjugated hyperbilirubinemia after birth. The proband developed into pleural effusions and ascites, pericardial thickening, intrahepatic and extrahepatic biliary duct dilatation, and enlarged gallbladder at age 50. Hepatocellular carcinoma occurred in the proband's brother at age 46. Seven compound defects of the ABCC2 gene [c.2414delG, p.(Ile1489Gly), p.(Thr1490Pro), and p.(Ile1491Gln)] and the UGT1A1 gene (c.-3279T>G, p.(Gly71Arg), and p.(Pro451Leu)) were identified in family members. Accumulation of pigment in hepatocytes characteristic of that in DJS was present in the proband and his brother. Expression of ABCC2 protein was markedly diminished in the patient's liver. Our results show a different genetic profile of DJS and GS in a Han Chinese family, indicating a more complex pattern of dual-hereditary jaundice among different populations. The present study illuminates the underpinnings of DJS and GS and extends the mutation profiles and phenotypes of these two syndromes in dual-hereditary jaundice.

Erzhiwan Ameliorates Restraint Stress- and Monobenzone-Induced Depigmentation in Mice by Inhibiting Macrophage Migration Inhibitory Factor and 8-Hydroxy-2-Deoxyguanosine.

Purpose: Vitiligo is an autoimmune disease that results in the loss of epidermal melanocytes. The treatments for patients with vitiligo remain lacking. Erzhiwan (EZW), a traditional Chinese Medicine composed of Ligustri Lucidi Fructus and Ecliptae Herba, was used to ameliorate depigmentation since ancient China. This study aims to investigate the effect of EZW on vitiligo-related depigmentation. Methods: A vitiligo-related depigmentation mouse model was induced by monobenzone and restraint stress. The experimental depigmentation mice were treated with EZW. Histological observation of skin was conducted. Cutaneous oxidative damage and inflammation were determined. A network pharmacology analysis was carried out. Results: EZW reduced depigmentation score (p<0.01), cutaneous inflammatory infiltration (p<0.01), and CD8α-positive expression (p<0.01), and increased cutaneous melanin content in experimental depigmentation mice. EZW reduced stress reaction in experimental depigmentation mice (p<0.01). EZW inhibited 8-hydroxy-2-deoxyguanosine (8-OHdG)-related DNA oxidative damage in the skin (p<0.05, p<0.01). In addition, EZW reduced cutaneous macrophage migration inhibitory factor (MIF)-CD74-NF-κB signaling (p<0.01). The network pharmacology analysis demonstrated that EZW regulated necroptosis, apoptosis, and FoxO signaling pathways in vitiligo. An in vitro experiment showed that the main ingredient of EZW, specnuezhenide, protected against monobenzone and MIF-induced cell death in HaCaT cells (p<0.01). Conclusion: EZW ameliorates restraint stress- and monobenzone-induced depigmentation via the inhibition of MIF and 8-OHdG signaling. The findings provide a data basis of an utilization of EZW in vitiligo.

Aroma volatile compounds from two fresh pineapple varieties in China.

Volatile compounds from two pineapples varieties (Tainong No.4 and No.6) were isolated by headspace solid phase microextraction (HS-SPME) and identified and quantified by gas chromatography-mass spectrometry (GC/MS). In the Tainong No. 4 and No. 6 pineapples, a total of 11 and 28 volatile compounds were identified according to their retention time on capillary columns and their mass spectra, and quantified with total concentrations of 1080.44 µg·kg(-1) and 380.66 µg·kg(-1) in the Tainong No.4 and No. 6 pineapples, respectively. The odor active values (OAVs) of volatile compounds from pineapples were also calculated. According to the OAVs, four compounds were defined as the characteristic aroma compounds for the Tainong No. 4 pineapple, including furaneol, 3-(methylthio)propanoic acid methyl ester, 3-(methylthio)propanoic acid ethyl ester and δ-octalactone. The OAVs of five compounds including ethyl-2-methylbutyrate, methyl-2-methylbutyrate, 3-(methylthio)propanoic acid ethyl ester, ethyl hexanoate and decanal were considered to be the characteristic aroma compounds for the Tainong No. 6 pineapple.

[Effect of different electrical current intensities of electroacupuncture preconditioning on cardiac function and macrophage polarization in mice with acute myocardial ischemia].

OBJECTIVE: To observe the effect of different intensities of electroacupuncture (EA) preconditioning on car-diac function and polarization state of macrophages in mice with acute myocardial ischemia (AMI), so as to explore its possible mechanism underlying improvement of AMI. METHODS: A total of 50 male C57BL/6J mice were randomly divided into sham ope-ration, AMI model, and EA pretreatment groups (0.5 mA, 1 mA, 3 mA subgroups), with 10 mice in each group/subgroup. The mice in the EA pretreatment groups were subjected to EA stimulation of bilateral "Neiguan"(PC6) with 0.5, 1.0 and 3 mA respectively and frequency of 2 Hz/15 Hz for 20 min, once a day, for 3 days. The acute myocardial ischemia model was established by ligating the anterior descending branch (ADB) of the left coronary artery, while the sham operation only had a surgical suture trans-passed below the ADB but without ligation. The myocardial infarction area was measured after TTC staining, and the cardiac function ï¼»left ventricular ejection fraction (EF), short-axis contraction rate (FS)ï¼½ was detected by using echocardiography. The M1 macrophages were labeled with CD11b+F480+CD206low, M2 macrophages were labeled with CD11b+F480+CD206high and detected by using flow cytometry, and the expression levels of myocardial interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α), Toll-like receptor-4 (TLR4) proteins were detected by using Western blot. RESULTS: Compared with the sham operation group, the model group had a significant increase in the infarction area (P<0.000 1), number of cardiac macrophages and percentage of M1 type macrophages (P<0.000 1), and the expression levels of myocardial IL-1ß, TNF-α, TLR4 proteins (P<0.001, P<0.01), and a remarkable decrease in the levels of EF, FS and the percentage of M2 type macrophages (P<0.000 1). In contrast to those of the model group, the area of myocardial infarction (P<0.000 1, P<0.01), expression levels of myocardial IL-1ß, TNF-α, TLR4 proteins (P<0.01, P<0.05, P<0.001) in the 0.5 mA, 1 mA and 3 mA groups, number of macrophages and percentage of M1 macrophages (P<0.05) in the 1 mA group were significantly decreased, while the levels of EF and FS (P<0.000 1, P<0.05, P<0.001) in the 3 EA groups, and percentage of M2 macrophage (P<0.05) in the 1 mA group were significantly increased. Comparison among the 3 EA groups displayed that the effects of 1 mA group were significantly superior to those of 0.5 and 3 mA groups in up-regulating EF and FS (P<0.01, P<0.001), and in down-regulating the area of infarct myocardium (P<0.01, P<0.000 1), and the expression of TLR4 protein (P<0.01), and 0.5 mA group in the expression of IL-1ß and TNF-α proteins (P<0.05). CONCLUSION: EA preconditioning with electrical current intensities of 0.5 mA, 1 mA and 3 mA can effectively reduce myocardial infarction size, improve cardiac function in mice with AMI, which may be related with its effects in reducing the number of cardiac macrophages and down-regulating the expression of myocardial IL-1ß, TNF-α and TLR4 proteins. The therapeutic effect of 1 mA is better than that of 0.5 and 3 mA.

Mitochondrial DNA haplogroup F confers genetic susceptibility to chronic HBV infection for the Yi nationality in Lijiang, China.

Mitochondria are essential for hepatitis B virus (HBV) infection. Moreover, the findings of our previous study indicate that host mitochondrial genetic factors are associated with chronic hepatitis B (CHB) for the Han Chinese. However, in terms of genetic heterogeneity, the impact of mitochondria on host susceptibility to HBV infection in ethnic minorities in China remains unclear. Here, a total of 7070 subjects who had visited the hospital between June 1, 2019, and April 31, 2020, were enrolled for seroprevalence of HBV infection investigation. A total of 220 individuals with CHB (CHBs) and 223 individuals with a trace of HBV infection (spontaneously recovered subjects, SRs) were analyzed for mitochondrial DNA (mtDNA) sequence variations and classified into respective haplogroups. Haplogroup frequencies were compared between CHBs and SRs. Among eight nationalities, Yi nationality patients had the highest HBsAg prevalence rate (27.9% [95% CI: 25.3%-30.5%]) and the lowest vaccination rate (4.9% [95% CI: 3.7%-6.2%]). After adjustment for age and gender, haplogroup F was a risk factor for CHB infection (P = 0.049, OR = 2.079, 95% CI = 1.002-4.31), while D4 had a significant negative correlation with the HBeAg-positive rate (P = 0.024, OR = 0.215, 95% CI = 0.057-0.816). Together with our previous study, the findings indicate that different nationalities have different genetic susceptibility to HBV infection.

Dickkopf-related protein 1/cytoskeleton-associated protein 4 signaling activation by Helicobacter pylori-induced activator protein-1 promotes gastric tumorigenesis via the PI3K/AKT/mTOR pathway.

BACKGROUND: Gastric cancer (GC) is a common malignant tumor with high incidence and mortality rates globally, especially in East Asian countries. Helicobacter pylori (H. pylori) infection is a significant and independent risk factor for GC. However, its underlying mechanism of action is not fully understood. Dickkopf-related protein (DKK) 1 is a Wnt signaling antagonist, and cytoskeleton-associated protein (CKAP) 4 is a newly identified DKK1 receptor. Recent studies found that the binding of DKK1 to CAKP4 mediated the procancer signaling of DKK1 inde-pendent of Wnt signaling. We hypothesize that H. pylori-induced activation of DKK1/CKAP4 signaling contributes to the initiation and progression of GC. AIM: To investigate the interaction of H. pylori infection, DKK1 and CAKP4 in GC, as well as the underlying molecular mechanisms. METHODS: RNA sequencing was used to identify differentially expressed genes (DEGs) between H. pylori-infected and uninfected primary GC cells. Gain- and loss-of-function experiments were performed to verify the H. pylori-induced upregulation of activator protein-1 (AP-1) in GC cells. A dual-luciferase reporter assay and co-immunoprecipitation were used to determine the binding of AP-1 to the DKK1 promoter and DKK1 to CKAP4. Western blotting and immunohistochemistry detected the expression of DKK1, CKAP4, and phos-phatidylinositol 3-kinase (PI3K) pathway-related proteins in GC cells and tissues. Functional experiments and tumorigenicity in nude mice detected malignant behavior of GC cells in vitro and in vivo. RESULTS: We identified 32 DEGs between primary GC cells with and without H. pylori infection, including JUN, fos-like antigen-1 (FOSL1), and DKK1, and confirmed that the three proteins and CKAP4 were highly expressed in H. pylori-infected GC cells, H. pylori-infected gerbil gastric tissues, and human GC tissues. JUN and FOSL1 form AP-1 to transcriptionally activate DKK1 expression by binding to the DKK1 promoter. Activated DKK1 bound to CKAP4, but not the most common Wnt coreceptor low-density lipoprotein receptor-related protein 5/6, to promote GC cell growth, colony formation, migration, invasion, and xenograft tumor growth in nude mice. All these effects were driven by activation of the PI3K/AKT/mammalian target of rapamycin (mTOR) pathway. Targeting the PI3K signaling pathway by LY294002 inhibited DKK1-mediated CKAP4/PI3K signaling activity and the malignant behavior of GC cells. CONCLUSION: H. pylori induces JUN and FOSL1 expression to form AP-1, which transcriptionally activates DKK1. Binding of DKK1 to KAKP4 contributes to gastric tumorigenesis via the PI3K/AKT/mTOR pathway.

Characteristic aroma compounds from different pineapple parts.

Characteristic aroma volatile compounds from different parts of cayenne pineapple were analyzed by headspace-solid phase microextraction (HS-SPME) and gas chromatography-mass spectrometry (GC/MS). The main volatile compounds were esters, terpenes, ketones and aldehydes. The number and content of aroma compounds detected in pulp were higher than those found in core. In pulp, the characteristic aroma compounds were ethyl 2-methylbutanoate, ethyl hexanoate, 2,5-dimethyl-4-hydroxy-3(2H)-furanone (DMHF), decanal, ethyl 3-(methylthio)propionate, ethyl butanoate, and ethyl (E)-3-hexenoate; while in core the main compounds were ethyl 2-methylbutanoate, ethyl hexanoate and DMHF. The highest odor units were found to correspond to ethyl 2-methylbutanoate, followed by ethyl hexanoate and DMHF. The odor units found for pulp were higher than those for core.

Electroacupuncture improves cardiac function and reduces infarct size by modulating cardiac autonomic remodeling in a mouse model of myocardial ischemia.

BACKGROUND: Sympathetic and parasympathetic nerve remodeling play an important role in cardiac function after myocardial ischemia (MI) injury. Increasing evidence indicates that electroacupuncture (EA) can regulate cardiac function by modulating the autonomic nervous system (ANS), but little is known about its effectiveness on neural remodeling post-MI. OBJECTIVES: To investigate the role of EA in ANS remodeling post-MI. METHODS: Adult male C57/BL6 mice were equally divided into the Control (Ctrl), MI and EA groups after generating the MI model by ligating the left anterior descending (LAD) coronary artery. Echocardiography and 2,3,5-triphenyltetrazolium (TTC) staining were employed to evaluate cardiac function and infarct size after EA treatment for five consecutive days. Serum norepinephrine (NE) levels were measured by ELISA to quantify sympathetic activation. Then, ANS remodeling was detected by immunohistochemistry (IHC), RT-qPCR, and Western blotting. RESULTS: Our preliminary findings showed that EA increased ejection fraction and fractional shortening and reduced infarct area after MI injury. Serum NE levels in the EA group were significantly decreased compared with those in the MI group. IHC staining results demonstrated that the density of growth associated protein (GAP)43 and tyrosine hydroxylase (TH) positive nerve fibers in the EA group were decreased with increased choline acetyltransferase (CHAT) and vesicular acetylcholine transporter (VACHT). Meanwhile, the results verified that mRNA and protein expression of GAP43 and TH were significantly inhibited by EA treatment in the MI mice, accompanied by elevated CHAT and VACHT. CONCLUSIONS: EA treatment could improve cardiac function and reduce infarct size by modulating sympathetic and parasympathetic nerve remodeling post-MI, thus helping the cardiac ANS reach a new balance to try to protect the heart from further possible injury.

Electroacupuncture preconditioning attenuates acute myocardial ischemia injury through inhibiting NLRP3 inflammasome activation in mice.

AIMS: Electro-acupuncture pretreatment (EAP) plays a protective role in myocardial ischemia (MI) injury. However, the underlying mechanism remains unclear. A growing body of evidence suggests postinfarction inflammatory response directly affects the remodeling of ventricular function. The purpose of this study was to investigate whether EAP alleviates MI through NLRP3 inflammasome inhibition. MATERIALS AND METHODS: We constructed an AMI model by ligating the left anterior descending (LAD) coronary artery after 3 days of EAP with C57BL/6 mice. Echocardiography and TTC staining were employed to evaluate cardiac function and infarct size after 24 h of ischemia. HE staining and immunohistochemistry were employed to determine inflammatory level. Then, inflammasome activation was detected by western blotting, and macrophage polarization and neutrophil infiltration were observed by flow cytometry. KEY FINDINGS: Our preliminary findings showed that EAP reduced the infarct area and increased fractional shortening (FS) and ejection fraction (EF) and decreased the degree of inflammation after AMI injury. Meanwhile, EAP inhibited the expression of NLRP3, cleaved caspase-1 and IL-1ß in ischemia myocardial tissue, companied by inhibiting the expression of F4/80+, CD11b+, CD206low macrophages and activated M2 macrophage, and decreasing Ly-6G+CD11b+ neutrophils in ischemia myocardial and spleen tissue. SIGNIFICANCE: EAP inhibits the activation of NLRP3 inflammasome, promotes M2 polarization of macrophages and reduces the recruitment of neutrophils in damaged myocardium, thereby decreases the infarct size and improves the cardiac function.

TRPV1 mediates cell death in rat synovial fibroblasts through calcium entry-dependent ROS production and mitochondrial depolarization.

Synoviocyte hyperplasia is critical for rheumatoid arthritis, therefore, potentially an important target for therapeutics. It was found in this work that a TRPV1 agonist capsaicin, and acidic solution (pH 5.5) induced increases in cytosolic calcium concentration ([Ca(2+)](c)) and reactive oxygen species (ROS) production in synoviocytes isolated from a rat model of collagen-induced arthritis. The increases in both [Ca(2+)](c) and ROS production were completely abolished in calcium-free buffer or by a TRPV1 antagonist capsazepine. Further experiments revealed that capsaicin and pH 5.5 solution caused mitochondrial membrane depolarization and reduction in cell viability; such effects were inhibited by capsazepine, or the NAD(P)H oxidase inhibitor diphenylene iodonium. Both capsaicin and pH 5.5 buffer induced apoptosis as shown by nuclear condensation and fragmentation. Furthermore, RT-PCR readily detected TRPV1 mRNA expression in the isolated synoviocytes. Taken together, these data indicated that TRPV1 activation triggered synoviocyte death by [Ca(2+)](c) elevation, ROS production, and mitochondrial membrane depolarization.

[Exogenous NO application effectively alleviates the allelochemical stress on cucumber root border cells caused by Eupatorium adenophorum extracts.] / 外源NOç¼“è§£ç´«èŒŽæ³½å °æå–ç‰©å¯¹é»„ç“œæ ¹è¾¹ç¼˜ç»†èƒžçš„åŒ–æ„Ÿèƒè¿«.

Using suspension cultures of cucumber (Cucumis sativus) cultivar Jinyou 35, we investigated the effects of allelochemical stresses induced by Eupatorium adenophorum extracts on root border cells (RBC), and the role of exogenous NO application in alleviation of the damage of root tips exposed to E. adenophorum extracts. The results showed that, 1000 mg·L-1 E. adenophorum extracts had significant damage to the cucumber root tip, resulting in severe tissue damage, exfoliated surface cells and irregular arrangement of inner cells, while those damages could be effectively alleviated by spraying exogenous NO. Compared with the control, E. adenophorum extracts (ZL) markedly reduced RBC numbers and survival rates by 54.5% and 97.2%, respectively, the RBC apoptosis rates were 12.3 times higher, the thicknesses of RBC adhesive layers were increased by 31.4%, and the root cap PME activities were markedly increased. Compared with the ZL treatment, exogenous NO application (ZN) significantly increased RBC numbers and survival rates by 72.4% and 146.0%, respectively, reduced the corresponding RBC apoptosis rates and the thicknesses of RBC adhesive layers by 30.7% and 15.0%, respectively, and inhibited the PME activities by 14.3% upon treatment for 72 hours. Our findings revealed that E. adenophorum extracts showed toxic effects on the cucumber RBC, resulting in cell apoptosis, abolishment of the RBC protection on root tips, and the destruction of root tip structure. Exogenous NO application, to some extent, could prevent the root tip and RBC from cell damage caused by E. adenophorum extracts.

Lymphomatosis cerebri: a rare variant of primary central nervous system lymphoma and MR imaging features.

BACKGROUND: Lymphomatosis cerebri (LC) is a rare variant of primary central nervous system lymphoma (PCNSL), characterized by diffuse infiltration without the formation of a discrete mass. The diagnosis of LC is a challenge because the imaging findings are atypical for lymphoma. The purpose of present study is to investigate MRI characteristics and clinical features of LC and potentially facilitate an early and accurate diagnosis of this often-missed disease. METHODS: Seven patients (average 44 years, 19-58 years) with LC proved basing on MRI and histology were retrospectively reviewed the clinical data and cerebral MR imaging findings. RESULTS: The common presenting symptoms were cognitive decline, behavioral disturbance, gait disturbance. All patients had both deep and lobar lesion distribution, and two of them had infratentorial involvement. Lack of contrast enhancement and subtle patchy enhanced pattern were observed in two and three patients, respectively. The remaining two patients presented multiple patchy enhancement. Most of the lesions were slightly hyperintense to normal brain on DWI as well as hyperintense on ADC maps. Three patients presented a pattern of marked decrease of NAA/Cr, increase of Cho/Cr, and two of the three cases showed increased Lip/Cr and Lac/Cr on MRS. CONCLUSIONS: We conclude that diffuse bilateral lesions especially in deep and lobar region including white and gray matter, without enhancement or with patchy enhancement, marked decrease of NAA/Cr and increase of Cho/Cr, and increased Lip/Cr and Lac/Cr are suggestive of LC. Prompt recognition of these imaging patterns may lead to early diagnosis of LC and brain biopsy with improved prognosis.

[Expression of BCL-10 protein and the relationship with API2-MALT1 fusion gene in extranodal marginal zone B-cell lymphoma of mucosa associated lymphoid tissue].

OBJECTIVE: To investigate the relationship of BCL-10 protein and API2-MALT1 fusion gene in MALT lymphoma. METHODS: Specimens from 86 cases of MALT lymphoma were studied by immunohistochemical staining for BCL-10. RT-PCR was used to detect the transcripts of API2-MALT1 fusion gene. RESULTS: In all 10 cases of Hashimoto thyroiditis only cytoplasmic BCL-10 expression in lymphoid cells was observed. In 86 MALT lymphoma cases, 42 cases (48. 8%) exhibited BCL-10 expression in both nucleus and cytoplasm. API2-MALT1 fusion gene was detected in 35 cases (40. 7%) of MALT lymphoma. BCL-10 nuclear expression was correlated with API2-MALT1 fusion gene transcript (r = 0. 374,P = 0. 000). CONCLUSION: BCL-10 nuclear expression is correlated with API2-MALT1 fusion gene expression in MALT lymphoma.

[Detection of API2-MALT1 fusion gene in extranodal B-cell lymphoma and its significance].

OBJECTIVE: To study the expression of API2-MALT1 mRNA in mucosa-associated lymphoid tissue (MALT) lymphoma, extranodal diffuse large B-cell lymphoma (DLBCL) and Hashimoto's thyroiditis, to investigate the expression pattern of API2-MALT1 variants, and to correlate the findings with the clinicopathologic features and prognosis. METHODS: Sixty-two cases of MALT lymphoma (10 from lung, 31 from stomach, 9 from intestine and 12 from thyroid), 32 cases of extranodal DLBCL (16 from stomach, 13 from intestine and 3 from thyroid), 8 cases of Hashimoto's thyroiditis and 5 cases of reactive lymph nodes hyperplasia as negative controls were collected. The expression of API2-MALT1 mRNA was studied in all cases by reverse transcriptase (RT)-polymerase chain reaction (PCR) and nested PCR. The 94 cases of lymphoma were subdivided into API2-MALT1-positive and API2-MALT1-negative groups. Among the patients, 78 were followed up for 6 to 120 months. The differences in clinicopathologic features and prognosis between the two groups were analyzed. RESULTS: API2-MALT1 transcripts were detected in 39 of the 94 lymphoma cases (with 28 cases being MALT lymphoma and 11 cases being extranodal DLBCL). mRNA expression was not detected in all cases of Hashimoto's thyroiditis and the negative controls. Two fusion gene variants, A1446-M1123 and A1446-M814 were found, and A1446-M1123 expression was more common. As for MALT lymphoma cases, the frequency of the fusion gene expression was lower in thyroid, when compared with that in lung, stomach and intestine. API2-MALT1-positive cases had tumors in an earlier stage with milder infiltration of cancer cells, lower relapse rate, and higher five-year survival rate. CONCLUSIONS: The expression of API2-MALT1 mRNA can be detected in both MALT lymphoma and extranodal DLBCL, but not in Hashimoto's thyroiditis. These cases tend to show a more indolent clinical course and better survival. The frequency of t (11; 18) (q21; q21) correlates with the primary sites of MALT lymphoma. The higher incidence of breakpoint at 1123 bp of MALT1 gene in Chinese people may be due to geographical variation.

[The frequency of API2-MALT1 fusion gene variants in MALT lymphoma and its correlation with apoptosis of MALT lymphoma].

OBJECTIVE: To investigate the frequency of different variant of API2-MALT1 fusion gene in extranodal marginal zone B-cell lymphoma of mucosa-associated tissue(MALT1) lymphoma and the correlation between API2-MALT1 transcript and apoptosis of MALT lymphoma. METHODS: The API2-MALT1 fusion transcripts were detected in 62 cases of MALT lymphoma by reverse transcription-polymeras chain reaction(RT-PCR) and Nested PCR. Five cases with reactive proliferation of lymph node were in use for negative control, and beta-actin was regarded as internal control; the apoptosis index, mRNA and protein of API2 were assayed in all samples by means of TUNEL, RT-PCR and immunohistochemistry respectively. RESULTS: API2-MALT1 transcript was detected in 28 of 62 cases with MALT lymphoma (45.16%). Two kinds of API2-MALT1 variants (A1446-M1123 and A1446-M814) were detected. Variant A1446-M1123 was detected more frequently as compared with A1446-M814. The frequency of API2-MALT1 transcript was lower in thyroid MALT lymphoma(1/12) but similar in pulmonary and gastrointestinal MALT lymphoma. In the group of API2-MALT1(+), the apoptosis index was higher and the API2 mRNA and protein were lower when compared against those in the group of API2MALT1(-). But no significant differences in the levels of apoptosis and API2 were observed between the group of A1446-M1123(+) and A1446-M814(+). CONCLUSION: API2-MALT1 transcript displayed variable frequency in MALT lymphomas of different sites. A1446-M1123 was noted to be probably the main type of API2-MALT1 variant in MALT lymphoma of Chinese. API2-MALT1 transcript was confirmed to be associated with the levels of apoptosis and API2 of MALT lymphoma.

[Expression of bcl-10 protein in MALT lymphoma].

OBJECTIVE: To investigate the significance of bcl-10 protein expression in extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) lymphoma. METHODS: Sixty-two cases of MALT lymphoma were reviewed and immunohistochemical studies for bcl-10 and Ki-67 were performed. RESULTS: Sixty out of the 62 cases studied (96.8%) were positive for bcl-10. Thirty-three (53.2%) showed bcl-10 expression in both the nuclei and cytoplasm, while 27 cases (43.6%) showed only cytoplasmic staining. The 10 cases with Hashimoto's thyroiditis demonstrated bcl-10 expression in the cytoplasm. The mean age of patients with bcl-10 nuclear expression (51.4 years old) was 5.2 years younger than those (56.6 years) without bcl-10 nuclear expression. The former category also showed a male predilection (male to female ratio = 19:14, in contrast to 10:19 in the latter category). The frequency of bcl-10 nuclear expression was lower in cases from thyroid but higher in cases from lung, stomach and intestine (P < 0.05). There was no statistically significant correlation between bcl-10 nuclear expression and clinical tumor stage (P > 0.05) or tumor cell morphology (P > 0.05). Amongst the 40 cases of gastrointestinal MALT lymphoma, bcl-10 nuclear expression correlated with extent of tumor involvement. The protein was expressed in 36.4% (4 out of 11 cases) of MALT lymphoma confined to mucosa or submucosa, 65.2% (15 out of 23 cases) of those invading down to muscularis propria or subserosa, and 100% (all 6 cases) of those extending beyond serosa (P < 0.05). There was no statistically significant difference in Ki-67 proliferative index between bcl-10-positive and bcl-10-negative groups (P < 0.05). Follow-up data were available in 52 patients (83.9%) and the five-year survival rate was no statistically significant difference in survival between bcl-10-positive (29 patients, 96.3%) and bcl-10-negative groups (23 patients, 66.4%, P > 0.05). CONCLUSIONS: Two expression patterns of bcl-10 protein were observed in MALT lymphoma: mixed nuclear-cytoplasmic and cytoplasmic only. The bcl-10 nuclear expression appears more important and correlates with anatomic site of tumor and extent of tumor involvement. Immunohistochemical detection of bcl-10 may carry some diagnostic and prognostic implications in assessment of MALT lymphoma.

[Detecting the API2-MALT1 fusion gene and its significance in Hashimoto's thyroiditis and thyroid lymphomas].

OBJECTIVE: To investigate the expression of API2-MALT1 fusion gene mRNA in Hashimoto's thyroiditis, thyroid MALT lymphoma and DLBCL, as well as the correlation between the expression and the pathogenesis and prognosis of two types of thyroid lymphoma. METHODS: Eight cases of Hashimoto's thyroiditis, 12 cases of MALT lymphoma and 3 cases of DLBCL in thyroid were selected and retrieved. API2-MALT1 transcripts were investigated in all cases by reverse transcript touchdown-PCR and nested PCR. The effect of API2-MALT1 fusion gene on the prognosis of thyroid lymphomas was assessed by analyzing the clinicopathological features and the follow-up findings. RESULTS: The variant 1446/1123 transcript was detected in 2 of thyroid lymphomas (1 MALT lymphoma, 1 DLBCL) but not in 8 cases of Hashimoto's thyroiditis and 5 cases of chronic lymphadenitis. Both of API2-MALT1-positive cases were accompanied with Hashimoto's thyroiditis, showing stage I E and no involvement of lymph node. CONCLUSION: The incidence of API2-MALT1 transcript in thyroid lymphomas was low; however, the correlation between API2-MALT1 fusion gene and the progression from Hashimoto's thyroiditis to lymphoma could not be excluded; the thyroid lymphoma with API2-MALT1 transcript seemed to develop on a more indolent clinical course.

Signal pathways involved in emodin-induced contraction of smooth muscle cells from rat colon.

AIM: To investigate the effects induced by emodin on single smooth muscle cells from rat colon in vitro, and to determine the signal pathways involved. METHODS: Cells were isolated from the muscle layers of Wistar rat colon by enzymatic digestion. Cell length was measured by computerized image micrometry. Intracellular Ca2+ ([Ca2+]i) signals were studied using the fluorescent Ca2+ indicator fluo-3 and confocal microscopy. PKCalpha distribution at rest state or after stimulation was measured with immunofluorescence confocal microscopy. RESULTS: (1) Emodin dose-dependently caused colonic smooth muscle cells contraction; (2) emodin induced an increase in intracellular Ca2+ concentration; (3) the contractile responses induced by emodin were respectively inhibited by preincubation of the cells with ML-7 (an inhibitor of MLCK) and calphostin C (an inhibitor of PKC); (4) Incubation of cells with emodin caused translocation of PKCalpha from cytosolic area to the membrane. CONCLUSION: Emodin has a direct contractile effect on colonic smooth muscle cell. This signal cascade induced by emodin is initiated by increased [Ca2+]i and PKCalpha translocation, which in turn lead to the activation of MLCK and the suppression of MLCP. Both of them contribute to the emodin-induced contraction.