Threatment Strategies for Recurrent Hepatocellular Carcinoma Patients: Ablation and its Combination Patterns.

With the development of guidance technology and ablation equipment, ablative procedures have emerged as important loco-regional alternatives to surgical resection for recurrent hepatocellular carcinoma (rHCC) patients. Currently, ablation modalities used in clinical practice mainly include radiofrequency ablation (RFA), microwave ablation (MWA), laser ablation (LA), cryoablation (CRA), high-intensity focused ultrasound (HIFU), and irreversible electroporation (IRE). Accumulated comparative data of ablation versus surgical resection reveal noninferior responses and outcomes but superior adverse effects. Moreover, studies demonstrate that ablation may serve as an excellent procedure for rHCC given its exact minimal invasiveness and immune modulation. We focus on the current status of ablation in clinical practice for rHCC and discuss new research in the field, including ablation combined with these other modalities, such as targeted therapy and immunotherapy.

A metal chaperone OsHIPP16 detoxifies cadmium by repressing its accumulation in rice crops.

Cadmium (Cd) is an environmentally polluted toxic heavy metal and seriously risks food safety and human health through food chain. Mining genetic potentials of plants is a crucial step for limiting Cd accumulation in rice crops and improving environmental quality. This study characterized a novel locus in rice genome encoding a Cd-binding protein named OsHIPP16, which resides in the nucleus and near plasma membrane. OsHIPP16 was strongly induced by Cd stress. Histochemical analysis with pHIPP16::GUS reveals that OsHIPP16 is primarily expressed in root and leaf vascular tissues. Expression of OsHIPP16 in the yeast mutant strain ycf1 sensitive to Cd conferred cellular tolerance. Transgenic rice overexpressing OsHIPP16 (OE) improved rice growth with increased plant height, biomass, and chlorophyll content but with a lower degree of oxidative injury and Cd accumulation, whereas knocking out OsHIPP16 by CRISPR-Cas9 compromised the growth and physiological response. A lifelong trial with Cd-polluted soil shows that the OE plants accumulated much less Cd, particularly in brown rice where the Cd concentrations declined by 11.76-34.64%. Conversely, the knockout oshipp16 mutants had higher levels of Cd with the concentration in leaves being increased by 26.36-35.23% over the wild-type. These results suggest that adequate expression of OsHIPP16 would profoundly contribute to Cd detoxification by regulating Cd accumulation in rice, suggesting that both OE and oshipp16 mutant plants have great potentials for restricting Cd acquisition in the rice crop and phytoremediation of Cd-contaminated wetland soils.

OsZIP11 is a trans-Golgi-residing transporter required for rice iron accumulation and development.

Iron (Fe) is a mineral nutrient necessary for plant growth and development. Whether the rice ZRT/IRT-like protein family metal transporter OsZIP11 is involved in Fe transport has not been functionally defined. The objective of the study is to figure out the essential role of the uncharacterized OsZIP11 played in rice growth, development, and iron accumulation, particularly in seeds. Transient subcellular location assays show that OsZIP11 was targeted to the trans-Golgi network. OsZIP11 was preferentially expressed in the rice tissues (or organs) at later flowering and seed development stages. Transcripts of OsZIP11 were significantly induced under Fe but not under zinc (Zn), copper (Cu) or manganese (Mn) deficiency. Yeast (Saccharomyces cerevisiae) transformed with OsZIP11 sequences displayed an active iron input which turned out that excessive iron accumulated in the cells. Knocking out OsZIP11 by CRISPR-Cas9 approach led to the attenuated rice growth and physiological phenotypes, depicting shorter plant height, reduced biomass, chlorosis (a symptom of lower chlorophyll concentration), and over-accumulation of malondialdehyde (complex representing the peroxidation of membrane lipids) in rice plantlets. The field trials demonstrated that OsZIP11 mutation impaired the capacity of seed development, with shortened panicle and seed length, compromised spikelet fertility, and reduced grain per plant or 1000-grain weight. Knocking out OsZIP11 also lowered the accumulation of iron in the brown rice by 48-51% compared to the wild-type. Our work pointed out that OsZIP11 is required for iron acquisition for rice growth and development.

Allergy-Related Sialodochitis: A Preliminary Cohort Study.

OBJECTIVES/HYPOTHESIS: To explore the clinically feasible diagnosis criteria and treatment outcomes of allergy-related sialodochitis (ARS). STUDY DESIGN: Prospective Cohort Study. METHODS: Ninety-six consecutive patients were enrolled by the following criteria: 1) recurrent swelling of ≥2 large salivary glands that lasted for ≥3 months; 2) with mucus plug exudations; 3) with atopic diseases; 4) ductal stenosis and/or ectasia. Sixty-four patients with elevation of peripheral blood eosinophil (PBE) and/or serum IgE level comprised group A (highly-suspected ARS group), while the remaining 32 comprised group B (patients without confirmed evidence of ARS). These patients were treated with interventional endoscopy. A chronic obstructive sialadenitis symptom (COSS) questionnaire was used to quantify the treatment outcomes. RESULTS: In group A, Serum IgE was elevated in 84.4% of patients and PBE was elevated in 34.4% of patients. Percentage of submandibular gland involvement was higher in group A than group B (48.4% vs. 18.8%). On sialograms, the snowflake changes of branch ducts were seen in higher percentage of group A compared with group B (59% vs. 35% for parotid glands, 27% vs. 8% for submandibular glands, respectively). Mucus plug smears showed abundant eosinophils in 14 group A patients. Biopsy of five group A patients revealed significant eosinophil infiltration around the main and interlobular ducts. During follow-up, the COSS scores were significantly decreased in both groups, and group B was improved better than group A. CONCLUSION: PBE and serum IgE are important diagnostic indexes of ARS. Mucus plug smear or histopathology verifies the diagnosis. Interventional endoscopy is helpful for ARS cases. LEVEL OF EVIDENCE: 3 Laryngoscope, 131:2030-2035, 2021.

Identification of a new function of metallothionein-like gene OsMT1e for cadmium detoxification and potential phytoremediation.

Cadmium (Cd) is a biologically non-essential and toxic heavy metal leaking to the environment via natural emission or anthropogenic activities, thereby contaminating crops and threatening human health. Metallothioneins (MTs) are a group of metal-binding proteins playing critical roles in metal allocation and homeostasis. In this study, we identified a novel function of OsMT1e from rice plants. OsMT1e was dominantly expressed in roots at all developmental stages and, to less extent, expressed in leaves at vegetative and seed filling stages. OsMT1e was mainly targeted to the nucleus and substantially induced by Cd exposure. Expression of OsMT1e in a yeast Cd-sensitive strain ycf1 conferred cellular tolerance to Cd, even though the ycf1 + OsMT1e cells accumulated more Cd than the control cells (ycf1 + pYES2). Both transgenic rice overexpressing (OX) and repressing OsMT1e by RNA interference (RNAi) were developed. Phenotypic analysis revealed that OsMT1e overexpression enhanced the rice growth concerning the increased shoot or root elongation, dry weight and chlorophyll contents, whereas the RNAi lines displayed a sensitive growth phenotype compared to wild-type. Assessment with 0.5, 2 and 10 µM Cd for two weeks revealed that the RNAi lines accumulated less Cd, while the OX lines had an increased Cd accumulation in root and shoot tissues. The contrasting Cd accumulation phenotypes between the OX and RNAi lines were further confirmed by a long-term study with 0.5 µM Cd for one month. Overall, the study unveiled a new function of OsMT1e in rice, which can be potentially used for engineering genotypes for phytoremediation or minimizing Cd in rice crops.

Developing a cadmium resistant rice genotype with OsHIPP29 locus for limiting cadmium accumulation in the paddy crop.

Widespread contamination of agricultural soil with toxic metals such as cadmium (Cd) is a major threat to crop production and human health. Metallochaperones are a unique class of proteins that play pivotal roles in detoxifying metallic ions inside cells. In this study, we investigated the biological function of an uncharacterized metallochaperone termed OsHIPP29 in rice plants and showed that OsHIPP29 resides in the plasma membrane and nucleus and detoxifies excess Cd and Zn. OsHIPP29 was primarily expressed in shoots during the vegetative stage and in leaf sheath and spikelet at the flowering stage. It can be differentially induced by excess Cd, Zn, Cu, Fe and Mn. To identify the function of OsHIPP29 in mediating rice response to Cd stress, we examined a pair of OsHIPP29 mutants, RNAi lines and transgenic rice overexpressing OsHIPP29 (OX) under Cd stress. Both mutant and RNAi lines are sensitive to Cd in growth as reflected in decreased plant height and dry biomass. In contrast, the OX lines showed better growth under Cd exposure. Consistent with the phenotype, the OX lines accumulated less Cd in both root and shoot tissues, whereas OsHIPP29 knockout led to higher accumulation of Cd. These results point out that expression of OsHIPP29 is able to contribute to Cd detoxification by reducing Cd accumulation in rice plants. Our work highlights the significance of OsHIPP29-mediated reduced Cd in rice plants, with important implications for further developing genotypes that will minimize Cd accumulation in rice and environmental risks to human health.

OsNHAD is a chloroplast membrane-located transporter required for resistance to salt stress in rice (Oryza sativa).

Salt stress is one of the major environmental factors limiting crop productivity. Although physiological and molecular characterization of salt stress response in plants has been the focus for many years, research on transporters for sodium ion (Na+) uptake, translocation and accumulation in plants, particularly in food crops like rice is limited. In this study, we functionally identified an uncharacterized sodium ion transporter named OsNHAD which encodes a putative Na+ / H+ antiporter in rice. Homology search shows its close relation to the Arabidopsis Na+/H+ antiporter AtNHD1 with 72.74% identity of amino acids. OsNHAD transcripts mainly express in leaves and are induced by Na+ stress. Confocal laser scanning microscopy analysis of OsNHAD::GFP fusion in tobacco leaves shows that OsNHAD resides in the chloroplast envelop. Knock-down of OsNHAD by RNA interference led to increased rice sensitivity to Na+, manifested by stunted plant growth, enhanced cellular damage, reduced PSII activity and changed chloroplast morphology. Mutation of OsNHAD also resulted in accumulation of more Na+ in chloroplasts and in shoots as well, suggesting that OsNHAD is involved in mediating efflux and detoxification of Na+ but does not affect K+ accumulation in plant cells. Complementation test reveals that OsNHAD was able to functionally restore the Arabidopsis mutant atnhd1-1 growth phenotype. These results suggest that OsNHAD possibly mediates homeostasis of sodium ions in the subcellular compartments and tissues of the plants when challenged to salt stress.

Treatment strategy of hilar and intraglandular stones in wharton's duct: A 12-year experience.

OBJECTIVES/HYPOTHESIS: To suggest a strategy for transoral removal of hilar and intraparenchymal submandibular stones. STUDY DESIGN: Retrospective case series. METHODS: Retrospective evaluation was performed for 514 consecutive patients with hilar and intraparenchymal submandibular stones treated via endoscopy-assisted surgery from January 2006 to June 2018. Three patients had bilateral stones. The stones were classified as: hilar (type I), posthilar (type II), intraparenchymal (type III), and multiple stones (type IV). RESULTS: The affected glands included 311 with type I, 84 with type II, 65 with type III, and 57 with type IV stones. Stones were successfully removed in 478 glands (92.5%, 478/517). Main treatment techniques included hilum ductotomy in 311 glands, intraparenchymal ductotomy in 68, submandibulotomy in 14, intraductal retrieval in 74, and hilum ductotomy accompanied by intraductal retrieval in 11. At a mean 40-months follow-up of 478 successful cases, clinical outcomes were good in 425, fair in 27, and poor in 26 glands. Postoperative sialograms in 75 stone-free patients were categorized as: type I, normal (n = 6); type II, ectasia or stenosis in the main duct and no persistent contrast on functional films (n = 44); type III, ectasia or stenosis in the main duct and mild contrast retention (n = 15); and type IV, poor shape of the main duct and evident contrast retention (n = 10). Postoperative sialometry of 32 patients revealed no significant differences of the gland function between the two sides. CONCLUSIONS: Appropriate use of various endoscopy-assisted approaches helps preserve the gland and facilitates recovery of gland function in patients with different depths of hilo-parenchymal submandibular stones. LEVEL OF EVIDENCE: 4 Laryngoscope, 130:2360-2365, 2020.

Three-dimensional radiographic features of ameloblastoma and cystic lesions in the maxilla.

OBJECTIVES: To characterize the radiographic features of maxillary ameloblastoma (AM), odontogenic keratocyst (OKC) and dentigerous cyst (DC) comparatively by using spiral CT and cone beam CT (CBCT). METHODS: Clinical records, histopathological reports, and nonenhanced spiral CT or CBCT images of 191 consecutive patients with primary maxillary AMs, OKCs, or DCs were retrospectively acquired, and radiographic features were analyzed. RESULTS: The study included 118 males and 73 females (age: 5-84 years). 72.0% of AMs and 84.3% of OKCs originated from the posterior maxilla, while 69.6% of DCs occurred in the anterior maxilla. Among 25 AMs, 44.0% were of desmoplastic type, with honey-combed appearance. 84.0% of AMs were circular or oval in shape, 84.0% expanded buccally, and 36.0% invade the nasal floor. Among 89 OKCs of 88 patients, 61.8% were circular or oval, 58.4% expanded buccally, 49.4% were dentigerous, 41.6% nearly filled the maxillary sinus, and 13.5% invaded the nasal floor. 93.7% (74/79) of DCs enveloped a single tooth, and the tooth-cyst relationship was centripetal in 35, eccentric in 30, and circumferential in 9. Moreover, 98.2% (55/56) of the cysts enveloping a supernumerary tooth were DCs, while 80.9% (38/47) of the cysts enveloping the third molar were OKCs. CONCLUSIONS: Maxillary AMs tend to grow with buccal expansion and invade the nasal floor, and DAs with honey-combed lobularity are common. Maxillary OKCs have variant shapes and tend to invaginate the maxillary sinus. The tooth-cyst relationship of dentigerous OKCs and DCs can be centripetal, eccentric, or circumferential.

Evaluation of Parotid Gland Function Before and After Endoscopy-Assisted Stone Removal.

PURPOSE: To quantify gland function before and after endoscopy-assisted lithectomy for patients with parotid stones and to analyze correlations among different evaluation modalities. MATERIALS AND METHODS: This study investigated 58 patients (27 men and 31 women) with a stone larger than 5 mm or multiple parotid stones who underwent successful endoscopy-assisted surgery at the authors' center from August 2007 through September 2017. Meticulous postoperative manipulations were administered routinely for 3 to 6 months to promote functional recovery of the affected gland. Gland function was evaluated preoperatively and 6 to 36 months (mean, 12 months) postoperatively by sialography, scintigraphy, and sialometry. Statistical analyses were conducted to quantify gland function recovery and to distinguish correlations among the 3 objective tests. RESULTS: Preoperative sialograms exhibited ductal ectasia at the stone site with ductal stenosis anterior to the stone (n = 53) or duct interruption at the stone site (n = 5). Postoperative sialograms of 45 patients without stones were categorized as approximately normal (type I; n = 17); showing ectasia or stenosis of the main duct without persistent contrast on the functional film (type II; n = 16); showing ectasia or stenosis of the main duct with mild contrast retention (type III; n = 6); or showing poor ductal shape with evident contrast retention (type IV; n = 6). Scintigraphy of 23 preoperative and 12 postoperative patients and sialometry of 24 preoperative and 12 postoperative patients indicated severe preoperative impairment and postoperative improvement of gland function. Postoperatively, although no relevant differences in saliva flow rate were found between the 2 sides, scintigraphy showed lower function of the affected gland compared with the control side. Statistical data showed positive correlations among the 3 methods. Sialography intuitively reflected the ductal shape, whereas sialometry and scintigraphy were more sensitive for evaluating gland function. CONCLUSION: For patients with parotid stones, minimally invasive endoscopic surgery and meticulous postoperative manipulations help preserve the glands and facilitate recovery of gland function. The 3 evaluating modalities have certain positive correlations.

Bifid variations of the mandibular canal: cone beam computed tomography evaluation of 1000 Northern Chinese patients.

OBJECTIVES: The aim of this study was to evaluate and quantify variations of bifid mandibular canals (BMCs) in a population of Northern China by using cone beam computed tomography (CBCT). STUDY DESIGN: CBCT images of 1000 consecutive patients were analyzed by using the NewTom proprietary software. BMCs were identified and classified on the basis of the Naitoh classification. Linear and angular measurements of BMCs were performed. Statistical analyses were conducted by using χ2 and Wilcoxon tests. RESULTS: BMCs were observed in 13.2% of 1000 patients and 8.4% of 2000 sides. The prevalence of BMCs was significantly lower in patients in the first 2 decades and in cases with a class II molar relationship. The retromolar canal (68.4%) was the most common type of BMC observed. No buccolingual canals were identified; however, 2 special canals were detected. A classification system of 3 subtypes of retromolar canals was suggested. On average, the beginning site of the branches from the opening of the main canal was at a distance of 8.1 mm. The mean diameter and length of BMCs were 2.1 mm and 12.6 mm, respectively. CONCLUSIONS: This study underlined the prevalence and characteristics of BMCs in a population of Northern China. Preoperative identification of BMCs with CBCT may help prevent postoperative complications.

Effect on intensity of treadmill running on learning, memory and expressions of cell cycle-related proteins in rats with cerebral ischemia.

OBJECTIVE: We discussed the intensity of treadmill running on learning, memory and expression of cell cycle-related proteins in rats with cerebral ischemia. METHOD: Eighty healthy male SD rats were randomly divided into normal group, model group, intensity I group and intensity II group, with 20 rats in each group. The four-vessel occlusion method of Pulsinelli (4-VO) was used to induce global cerebral ischemia. Brain neuronal morphology was observed by hematoxylin-eosin (HE) staining at 3h, 6h, 24h and 48h after modeling, respectively. Hippocampal expressions of cyclin A and cyclin E were detected by immunohistochemistry. At 48h after modeling, the learning and memory performance of rats was tested by water maze experiment. RESULT: Compared with the normal group, the other three groups had a significant reduction in surviving neurons, prolonging of escape latency and decreased number of passes over the former position of the platform (P<0.05). The number of surviving neurons and the number of passes over the former position of the platform were obviously lower in the model group than in intensity I group (P<0.05), but significantly higher compared with intensity II group (P<0.05). Escape latency of the model group was obviously prolonged as compared with intensity I group (P<0.05), but much shorter than that of intensity II group (P<0.05). Compared with the normal group, the expressions of cyclin A and cyclin E were significantly upregulated at different time points after modeling (P<0.05). The expression of the model group was higher than that of intensity I group, but lower than that of intensity II group (P<0.05). CONCLUSION: Moderate intensity of treadmill running can help protect brain neurons and improve learning and memory performance of rats with global cerebral ischemia. But high intensity of treadmill running has a negative impact, possibly through the regulation of cell cycle-related proteins in ischemia/reperfusion injury.

Hippocampal mitogen-activated protein kinase activation is associated with intermittent hypoxia in a rat model of obstructive sleep apnea syndrome.

Obstructive sleep apnea syndrome (OSAS), characterized by intermittent hypoxia/re­oxygenation, may impair the cerebral system. Although mitogen­activated protein kinase (MAPK) signaling was observed to have a key role in hypoxia­induced brain injury, the intracellular events and their underlying mechanisms for intermittent hypoxia/re­oxygenation-associated damage to hippocamal MAPKs, including extracellular signal­regulated kinase (ERK)1/2, P38MAPK and c­Jun N­terminal kinase (JNK) remain to be elucidated and require further investigation. A total of five rats in each sub­group were exposed to intermittent hypoxia or continued hypoxia for 2, 4, 6 or 8 weeks. Histological, immunohistochemical and biological analyses were performed to assess nerve cell injury in the hippocampus. Surviving CA1 pyramidal cells were identified by hematoxylin and eosin staining. The levels of phosphorylated ERK1/2, P38MAPK and JNK were detected by western blotting. B­cell lymphoma 2 (Bcl­2) and Bcl­2­associated X protein (Bax) in neural cells were examined by immunohistochemistry. The malondialdehyde (MDA) contents and superoxide dismutase (SOD) activities were measured by thiobarbituric acid and xanthine oxidation methods, respectively. Under continued hypoxia, the levels of phospho­ERK1/2 peaked at the fourth week and then declined, whereas phospho­P38MAPK and JNK were detected only in the late stages. By contrast, under intermittent hypoxia, ERK1/2, P38MAPK and JNK were activated at all time-points assessed (2, 4, 6 and 8 weeks). The levels of phospho­ERK1/2, P38MAPK and JNK were all higher in the intermittent hypoxia groups than those in the corresponding continued hypoxia groups. Bcl­2 was mainly increased and reached the highest level at six weeks in the continued hypoxia group. Of note, Bcl­2 rapidly increased to the peak level at four weeks, followed by a decrease to the lowest level at the eighth week in the intermittent hypoxia group. Bax was generally increased at the late stages under continued hypoxia, but increased at all time-points under the intermittent hypoxia conditions. The two types of hypoxia induced an increase in the MDA content, but a decrease in SOD activity. Marked changes in these two parameters coupled with markedly reduced surviving cells in the hippocampus in a time­dependent manner were observed in the intermittent hypoxia group in comparison with the continued hypoxia group. OSAS­induced intermittent hypoxia markedly activated the MAPK signaling pathways, which were triggered by oxidative stress, leading to abnormal expression of downstream Bcl­2 and Bax, and a severe loss of neural cells in the hippocampus.

Adenovirus mediated BIMS transfer induces growth supression and apoptosis in Raji lymphoma cells.

OBJECTIVE: To transfer pro-apoptotic BIM directly into tumor cells bypass the complicated biological processes of BIM activation so as to reverse the chemoresistance of cancer cells. METHODS: BIMS was specifically amplified from HL-60 cells by RT-PCR, confirmed to be correct by sequencing and cloned into shuttle vector pAdTrack-CMV carrying a green fluorescence protein gene to generate a recombinant plasmid pAdTrack-CMV-BIMS. This plasmid and adenovirus backbone plasmid pAdEasy-1 were linearized and electroporated into E.coli BJ5183 host bacteria to mediate homologous recombination. The positive clone was identified by restrict endonuclease digestion. The recombinant pAdEasy-CMV-BIMS was transferred into HEK293 cells for packaging and amplification. The successful construction of recombinant human BIMS adenovirus (Ad-BIMS) was demonstrated by Western blot. To test whether Ad-BIMS has the capability of inducing apoptosis of tumor cells, Ad-BIMS was used to infect GC resistant Burkitt lymphoma Raji cells. RESULTS: After infected for 2-5 days, BIMS expression in Raji cells was detected by RT-PCR and Western blot. The significant growth retardation and apoptosis of Raji cells were also observed by MTT and flow cytometry. CONCLUSION: These results indicated that BIMS might be a potential candidate of gene therapy for chemoresistant tumor cells.

[Clinical analysis of Epstein-Barr virus-associated hemophagocytic syndrome in children].

The primary infection of Epstein-Barr virus (EBV) may results in hemophagocytic syndrome, known as EBV-associated hemophagocytic syndrome (EBV-AHS), but the clinical risk factors complicating this fatal disease in children with infectious mononucleosis (IM) are unknown. The aim of this study was to identify clinical features of EBV-AHS and to evaluate the curative effect of HLH-2004 protocol. The clinical and laboratory data of 644 IM children including 27 children developed into EBV-AHS and 43 HPS children associated with other diseases were retrospectively analyzed and logistic regression was used to identify the clinical risk factors complicating EBV-AHS. The results showed as follows: (1) the prevalence of EBV-AHS in IM children was 4.2% (27/644), and the prevalence in group aged younger than 3 years was higher than in other age groups. The incidence age of EBV-AHS was significantly younger than that of other HPS patients; (2) Liver function damage of group aged older than 7 years was much more severe in HPS patients. (3) Compared with other HPS patients, male patients were more common and liver function damage was severe in EBV-AHS patients, especially in the patients aged at 2 years or younger. (4) The fatality rate in the EBV-AHS patients was 37.0% (10/27). (5)After treatment with HLH-2004 protocol, the fatality rate in patients with EBV-AHS decreased from 50.0% to 18.2%, the overall survival (OS) of 3 years significantly increased (P = 0.032). It is concluded that IM is a benign self-limited disease, of which only about 4.2% patients will develop into EBV-AHS. Clinical risk factors identified in this study may be helpful for early diagnosis of IM children with complicated EBV-ASH, the HLH-2004 protocol can obviously improve prognosis of EBV-HPS.

Pro-apoptotic protein BIM in apoptosis of glucocorticoid-sensitive and -resistant acute lymphoblastic leukemia CEM cells.

Although glucocorticoids (GCs) have been used to treat acute lymphoblast leukemia (ALL) for decades, the mechanisms of GC sensitivity and resistance in ALL cells are poorly understood. This study investigated the role and mechanisms of pro-apoptotic protein BIM in apoptosis of GC-sensitive and- resistant ALL cells. The dramatic apoptosis was observed in GC-sensitive CEM-C7 cells after incubated with DEX for 48 h, while not in GC-resistant CEM-C1 cells. The significant up-regulation of BIM in CEM-C7 cells induced by DEX was also observed, but no up-regulation of BIM was detected in DEX-induced CEM-C1 cells. When treated with DEX plus RU486, a glucocorticoid receptor blocker, the apoptosis and BIM expression of CEM-C7 cells were canceled. P38MAPK-blocking pharmacon SB203580 also significantly inhibited the up-regulation of BIM in CEM-C7 cells. These suggested that the absence of BIM up-regulation is one of the important mechanisms of GC resistance, GC-GR conjugation is indispensible in both GC-induced apoptosis and up-regulation of BIM, and p38 MAPK signal pathway is also involved in this process.

[Clinical risk factors for Epstein-Barr virus-associated hemophagocytic syndrome in children with infectious mononucleosis].

OBJECTIVE: To compare the clinical features of infectious mononucleosis (IM) and Epstein-Barr virus (EBV)-associated hemophagocytic lymphohistiocytosis (EBV-AHS) and identify the clinical risk factors in IM patients complicated with EBV-AHS. METHOD: A retrospective study was carried out to analyze the clinical and laboratory data of 414 IM and 16 EBV-AHS children from January, 2000 to April, 2006. Then Logistic regression was used to identify the risk factors for progression to EBV-ASH. RESULTS: (1) The incidence of EBV-AHS among the IM children was 3.72% (16/430). There were significant differences between EBV-ASH and IM children in duration of fever (20 days vs. 7 days, P < 0.001), the peaks of fever (40.0 degrees C vs. 39.0 degrees C, P < 0.001), the degree of hepatomegaly (3.5 cm vs 2.0 cm below costal arch, P < 0.05) and splenomegaly (2.75 cm vs. 1.0 cm below costal arch, P < 0.05), while the incidence of isthmitis in EBV-AHS patients was markedly lower than that of IM patients (37.5% vs. 91.1%, P < 0.01). (2) Pancytopenia was often observed in EBV-AHS patients and significant differences between two groups were found in median of leukocytes (3.1 x 10(9)/L vs. 12.8 x 10(9)/L, P < 0.001), median of neutrophils (0.53 x 10(9)/L vs. 3.17 x 10(9)/L, P < 0.001), mean of hemoglobin (80 g/L vs. 120 g/L, P < 0.001) and median of platelet (27.5 x 10(9)/L vs. 183 x 10(9)/L, P < 0.001). (3) Hepatic derangement evidenced by elevated serum enzymes, hyperbilirubinemia and hypoalbuminemia in EBV-ASH children was much more severe than that in IM children, especially LDH level (2128.5 U/L vs. 445 U/L, P < 0.001) and AST level (489 U/L vs. 59 U/L, P < 0.001). (4) The clinical risk factors for IM patients progressing to EBV-ASH were lasting fever >/= 10 days (OR = 8.097, P = 0.008), LDH > 1000 U/L (OR = 7.998, P = 0.033), hypo-albuminemia (albumine < 35 g/L, OR = 7.838, P = 0.038), neutrophils < 1.5 x 10(9)/L (OR = 7.587, P = 0.022) and Plt < 100 x 10(9)/L (OR = 7.190, P = 0.027). The mortality of EBV-AHS in the patients was 50.0% (8/16). CONCLUSION: Most of IM children clinically manifest self-limited process, but about 3.72% of whom may progress to fatal EBV-ASH. The clinical risk factors for EBV-AHS are lasting fever > 10 days, LDH > 1000 U/L, hypoalbuminemia, neutropenia and Plt < 100 x 10(9)/L. EBV-ASH is an extremely dangerous state with high mortality. Repeated bone marrow examinations are helpful for diagnosis in time.

[Role of pro-apoptotic protein Bim in mouse thymocytes apoptosis induced by dexamethasone].

OBJECTIVE: To explore the role of pro-apoptotic protein Bim in the mouse thymocyte apoptosis induced by dexamethasone (DEX). METHODS: 1 micromol/L of dexamethasone was used to induce the apoptosis of BALB/c mouse thymocyte cultured in vitro. The thymocyte apoptosis was detected by transmission electron microscope, Annexin V-FITC/PI flow cytometry and DNA agarose gel electrophoresis. The RT-PCR method was applied to the semi-quantitative analysis of Bim mRNA expression induced by 1 micromol/L DEX at different time point and by different final concentration dexamethasone used for 1 h. RESULTS: (1) With 1 micromol/L DEX used to cell incubation for 4 h, the mouse thymocyte showed the morphological characteristics of apoptosis and the typical DNA ladder on electrophoresis gel. The analyses of Annexin V-FITC /PI flow cytometry showed that the thymocyte apoptosis rates at different time point (2 h, 4 h, 8 h) were significantly higher in dexamethasone treated group than in control group. (2) With 1 micromol/L DEX used to cell incubation for 1/2 hr, the expressions of three Bim mRNAs in mouse thymocyte began to increase and the BimL expression was predominant among three mRNA isoforms. The peak values of BimL and BimEL mRNA expression appeared at 1 h and 11 h which were earlier to dexamethasone treated group than control group, and the BimL and BimEL expression levels at different time point (except at 0 h and 24 h) were significantly increased (P < 0.05). The peak value of BimS mRNA expression of dexamethasone treated group was 4 hours earlier than that of control group, but there was no significant difference between two groups (P > 0. 05). (3) When the mouse thymocyte was incubated with various concentration (0, 10(-2),10(-1), 1, 10 micromol/L) DEX for 1 h, there was no significant percentage difference occurring between living and apoptotic cell. While the total of Bim mRNA expression was increasing in a concentration-dependent manner, the mRNA increase of BimL isoform was still the most predominant among three mRNA isoforms. CONCLUSION: The pro-apoptotic protein Bim plays an important role in mouse thymocyte apoptosis induced by dexamethasone, and the BimL mRNA is the predominant isoform in this process. The increase of Bim mRNA expression appears in very early stage of thymocyte apoptosis and both time- and dose-dependent.

[Research on A23187 inducing HL-60 cells to differentiate into dendritic cells].

OBJECTIVE: To investigate the method for calcium mobilization inducing rapidly to produce the dendritic cells (DC) from myeloid leukemia cells. METHODS: HL-60 cells were cultured with calcium ionophore A23187 (45-720 ng/mL) or plus rhIFN-gamma (1000 U/mL) for 20-96 hours. The morphologic features of cells were observed under inverted microscope and scanning electron microscope, when the cell phenotypes of HL-60 treated with A23187 were determined by flow cytometry. The proliferation of allogeneic human T cells was tested by mixed lymphocyte reaction (Allo-MLR). RESULTS: After treated with A23187 (180 ng/mL) or plus rhIFN-gamma (1000 U/mL) for 20 hours, some of HL-60 cells were found to have the dendritic appearance on cell surface; and the CD83 as a characteristic marker of mature DCs, co-stimulating molecules CD80 and CD86 got an up-regulated expression. When HL-60 cells were cultured with A23187 for 48 hours, the expression of CD83 began going down. A large number of cells with typical dendritic appearance were observed after cultured with A23187 for 72 hours, and the expressions of CD80 and CD86 were up-regulated continuously. Allo-MLR revealed that DCs derived from HL-60 cells treated with A23187 or plus rhIFN-gamma could stimulate the proliferation of allogeneic human T cells. CONCLUSION: The calcium ionophore A23187 can induce the HL-60 cells into mature DCs. This study suggests that calcium mobilization for cell differentiation may be an alternative way to rapidly acquire DCs.