Excessive iron deposition in root apoplast is involved in growth arrest of roots in response to low pH.

The rhizotoxicity of protons (H+) in acidic soils is a fundamental constraint that results in serious yield losses. However, the mechanisms underlying H+-mediated inhibition of root growth are poorly understood. In this study, we revealed that H+-induced root growth inhibition in Arabidopsis depends considerably on excessive iron deposition in the root apoplast. Reducing such aberrant iron deposition by decreasing the iron supply or disrupting the ferroxidases LOW PHOSPHATE ROOT 1 (LPR) and LPR2 attenuates the inhibitory effect of H+ on primary root growth efficiently. Further analysis showed that excessive iron deposition triggers a burst of highly reactive oxygen species, consequently impairing normal root development. Our study uncovered a valuable strategy for improving the ability of plants to tolerate H+ toxicity by manipulating iron availability.

Ultrasound-based artificial intelligence in gastroenterology and hepatology.

Artificial intelligence (AI), especially deep learning, is gaining extensive attention for its excellent performance in medical image analysis. It can automatically make a quantitative assessment of complex medical images and help doctors to make more accurate diagnoses. In recent years, AI based on ultrasound has been shown to be very helpful in diffuse liver diseases and focal liver lesions, such as analyzing the severity of nonalcoholic fatty liver and the stage of liver fibrosis, identifying benign and malignant liver lesions, predicting the microvascular invasion of hepatocellular carcinoma, curative transarterial chemoembolization effect, and prognoses after thermal ablation. Moreover, AI based on endoscopic ultrasonography has been applied in some gastrointestinal diseases, such as distinguishing gastric mesenchymal tumors, detection of pancreatic cancer and intraductal papillary mucinous neoplasms, and predicting the preoperative tumor deposits in rectal cancer. This review focused on the basic technical knowledge about AI and the clinical application of AI in ultrasound of liver and gastroenterology diseases. Lastly, we discuss the challenges and future perspectives of AI.

Does delaying ureteral stent placement lead to higher rates of preoperative acute pyelonephritis during pregnancy?

BACKGROUND: Pregnancy with renal colic may cause pyelonephritis, decreased renal function, systemic infection and even shock in pregnant women, and cause premature birth and other adverse pregnancy outcomes. When surgery is necessary, the relationship between timing of the operation and the outcome of the mother and child are not known. AIM: To investigate the association between time to ureteral stent placement and clinical outcomes of patients with renal colic during pregnancy. METHODS: In this retrospective study, pregnant women with renal colic who underwent surgery were studied. Maternal preoperative acute pyelonephritis (PANP), pregnancy outcome, and length of hospital stay (LOS) were compared between the two groups. RESULTS: 100 patients were included in the analysis, median age was 30 years. Median time to ureteral stent placement was 48 h (interquartile range, 25-96 h), and 32 patients (32%) were diagnosed with PANP. PANP was closely related to hospitalization costs, re-admission to the hospital due to urinary tract infection after surgery and premature delivery. Multivariate analysis found that stone location and time from pain to admission were related to PANP. CONCLUSION: Both early and delayed surgery are safe and effective for the treatment of renal colic during pregnancy. Early surgery may be superior to a delayed procedure due to shorter LOS. For pregnant patients with renal colic, delayed surgery within 48 h is not related to the clinical outcome of the mother and child. However, the time from pain to hospital admission was related to PANP.

The role of fibrosis index FIB-4 in predicting liver fibrosis stage and clinical prognosis: A diagnostic or screening tool?

This review evaluates the ability of the fibrosis index based on four factors (FIB-4) identifying fibrosis stages, long-time prognosis in chronic liver disease, and short-time outcomes in acute liver injury. FIB-4 was accurate in predicting the absence or presence of advanced fibrosis with cut-offs of 1.0 and 2.65 for viral hepatitis B, 1.45 and 3.25 for viral hepatitis C, 1.30 (<65 years), 2.0 (≥65 years), and 2.67 for non-alcoholic fatty liver disease (NAFLD), respectively, but had a low-to-moderate accuracy in alcoholic liver disease (ALD) and autoimmune hepatitis. It performed better in excluding fibrosis, so we built an algorithm for identifying advanced fibrosis by combined methods and giving work-up and follow-up suggestions. High FIB-4 in viral hepatitis, NAFLD, and ALD was associated with significantly high hepatocellular carcinoma incidence and mortality. Additionally, FIB-4 showed the ability to predict high-risk varices with cut-offs of 2.87 and 3.91 in cirrhosis patients and predict long-term survival in hepatocellular carcinoma patients after hepatectomy. In acute liver injury caused by COVID-19, FIB-4 had a predictive value for mechanical ventilation and 30-day mortality. Finally, FIB-4 may act as a screening tool in the secondary prevention of NAFLD in the high-risk population.

Upregulation of the checkpoint protein CHFR is associated with tumor suppression in pancreatic cancers.

The checkpoint with forkhead-associated (FHA) domain and RING-finger (CHFR) protein was identified as a cell cycle checkpoint protein and E3 ubiquitin ligase. In the present study, the potential functions of CHFR in pancreatic cancer were investigated. CHFR expression was measured in five pancreatic cancer cell lines by reverse transcription- quantitative polymerase chain reaction and western blotting. Capan-1 cells stably expressing CHFR were established by lentiviral vector transfection. Cell proliferation was assessed using Cell Counting Kit-8, and cell migration/invasion assay was determined using Transwell assays. Cell cycle and apoptosis induced by gemcitabine or docetaxel were evaluated using flow cytometry. CHFR expression levels were also evaluated in pancreatic ductal adenocarcinoma (PDAC) tumor samples as well as adjacent non-tumor tissues by immunohistochemistry. The significance of CHFR expression was determined, with respect to clinicopathological features and overall survival. Overexpression of CHFR in Capan-1 cells led to a decreased proliferative rate and reduced cell migration and invasion abilities. Results also indicated an increase in G1 phase cells in Capan-1 cells overexpressing CHFR. Docetaxel-induced apoptosis was inhibited in Capan-1 cells with CHFR-overexpression. A reduction in CHFR expression was detected in 51.9% of patients with PDAC, which significantly correlated with later T-stage. The results show CHFR functions as a tumor suppressor in pancreatic cancer, suggests its potential role in controlling the cell cycle of pancreatic cancer cells; however, CHFR overexpression is not a favorable factor in apoptosis induced by docetaxel.

Mutagenic and chemical analyses provide new insight into enzyme activation and mechanism of the type 2 iron-sulfur l-serine dehydratase from Legionella pneumophila.

The crystal structure of the Type 2 l-serine dehydratase from Legionella pneumophila (lpLSD), revealed a "tail-in-mouth" configuration where the C-terminal residue acts as an intrinsic competitive inhibitor. This pre-catalytic structure undergoes an activation step prior to catalytic turnover. Mutagenic analysis of residues at or near the active site cleft is consistent with stabilization of substrate binding by many of the same residues that interact with the C-terminal cysteine and highlight the critical role of certain tail residues in activity. pH-rate profiles show that a residue with pK of 5.9 must be deprotonated and a residue with a pK of 8.5 must be protonated for activity. This supports an earlier suggestion that His 61 is the likely catalytic base. An additional residue with a pK of 8.5-9 increases cooperativity when it is deprotonated. This investigation also demonstrates that the Fe-S dehydratases convert the enamine/imine intermediates of the catalytic reaction to products on the enzyme prior to release. This is in contrast to pyridoxyl 5' phosphate based dehydratases that release an enamine/imine intermediate into solution, which then hydrolyzes to produce the ketoamine product.

Silencing pancreatic adenocarcinoma upregulated factor (PAUF) increases the sensitivity of pancreatic cancer cells to gemcitabine.

Pancreatic adenocarcinoma upregulated factor (PAUF) is a new oncogene that activates signaling pathways that play a critical role in resistance to gemcitabine. We thus speculated that PAUF also plays a role in resistance to gemcitabine of pancreatic cancer cells. We established BxPC-3 cell lines with stable PAUF knockdown (BxPC-3_shPAUF) and controls (BxPC-3_shCtrl) and evaluated sensitivity to gemcitabine in vitro by MTT and flow cytometry. We established a xenograft model of human pancreatic cancer to examine PAUF function in gemcitabine resistance in vivo. Gene chip microarrays were performed to identify differentially expressed genes in BxPC-3_shPAUF and BxPC-3_shCtrl cells. Silencing PAUF increased the sensitivity of BxPC-3 cells to gemcitabine in vitro and in vivo. PAUF-knockdown BxPC-3 cell lines treated with gemcitabine showed increased proliferation inhibition and apoptosis compared with controls. Gemcitabine exhibited a more pronounced effect on reduction of BxPC-3_shPAUF tumors than BxPC-3_shCtrl tumors. Terminal deoxynucleotidyl transferase dUTP Nick-End Labeling (TUNEL) assays confirmed a significantly higher apoptotic rate of BXPC-3_shPAUF tumors compared with BXPC-3_shCtrl tumors. Gene array showed that PAUF function in gemcitabine sensitivity might involve MRP2, MRP3, MDR1, PIK3R1, and NFkB2 genes. PAUF could be considered as a key molecular target for sensitizing pancreatic cancer cells to gemcitabine.

Autologous sclera-muscle flaps technique in evisceration with hydroxyapatite implantation.

AIM: To provide superior cosmetic results and reduce complications, unlike traditional evisceration coupled with implant insertion technique and its modifications, we have developed a novel and simple technique for anophthalmic patients. METHODS: All patients who underwent the scleral-muscle flaps procedure in evisceration with the placement of hydroxyapatite implant were included in the study. Main outcome measures were complications such as exposure, infection, chemosis, conjunctival inclusion cysts, granulomas. Meanwhile, implant motility was indirectly measured and the results were collected and analyzed. RESULTS: A total of twenty-eight patients were enrolled in the study. Eighteen were men (64.29%) and ten were women (35.71%). Ages ranged from 18 to 65y (mean age, 32 years old). Mean follow-up was 12.32mo (range, 9-16mo). All patients received a hydroxyapatite implant. The average diameter of the implant was 19.29±1.36 mm (range, 18-22 mm). Minor complications occurred in 3 patients, and a major complication was observed in 1 patient. Mean motility were 11.04±1.45 mm horizontally (range, 7-14 mm) and 8.57±1.50 mm vertically (range, 5-12 mm). CONCLUSION: The sclera-muscle flaps technique in evisceration with hydroxyapatite implantation is simple and practical that eases the surgical procedure, enables a proper size hydroxyapatite implantation, distinctively reduces complications and provides superior surgery results, especially the motility of the implant.

Discovery and structural optimization of 1-phenyl-3-(1-phenylethyl)urea derivatives as novel inhibitors of CRAC channel.

AIM: Ca(2+)-release-activated Ca(2+) (CRAC) channel, a subfamily of store-operated channels, is formed by calcium release-activated calcium modulator 1 (ORAI1), and gated by stromal interaction molecule 1 (STIM1). CRAC channel may be a novel target for the treatment of immune disorders and allergy. The aim of this study was to identify novel small molecule CRAC channel inhibitors. METHODS: HEK293 cells stably co-expressing both ORAI1 and STIM1 were used for high-throughput screening. A hit, 1-phenyl-3-(1-phenylethyl)urea, was identified that inhibited CRAC channels by targeting ORAI1. Five series of its derivatives were designed and synthesized, and their primary structure-activity relationships (SARs) were analyzed. All derivatives were assessed for their effects on Ca(2+) influx through CRAC channels on HEK293 cells, cytotoxicity in Jurkat cells, and IL-2 production in Jurkat cells expressing ORAI1-SS-eGFP. RESULTS: A total of 19 hits were discovered in libraries containing 32 000 compounds using the high-throughput screening. 1-Phenyl-3-(1-phenylethyl)urea inhibited Ca(2+) influx with IC50 of 3.25±0.17 µmol/L. SAR study on its derivatives showed that the alkyl substituent on the α-position of the left-side benzylic amine (R1) was essential for Ca(2+) influx inhibition and that the S-configuration was better than the R-configuration. The derivatives in which the right-side R3 was substituted by an electron-donating group showed more potent inhibitory activity than those that were substituted by electron-withdrawing groups. Furthermore, the free N-H of urea was not necessary to maintain the high potency of Ca(2+) influx inhibition. The N,N'-disubstituted or N'-substituted derivatives showed relatively low cytotoxicity but maintained the ability to inhibit IL-2 production. Among them, compound 5b showed an improved inhibition of IL-2 production and low cytotoxicity. CONCLUSION: 1-Phenyl-3-(1-phenylethyl)urea is a novel CRAC channel inhibitor that specifically targets ORAI1. This study provides a new chemical scaffold for design and development of CRAC channel inhibitors with improved Ca(2+) influx inhibition, immune inhibition and low cytotoxicity.

[Analysis of cell morphology and immunophenotypic characteristics in 47 cases of multiple myeloma].

OBJECTIVE: This study was to investigate the cell morphology and cell immune phenotypic characteristics in patients with multiple myeloma (MM). METHODS: The flow cytometry with multiparametric direct immunofluorescence technique, and CD45/SSC and CD38(+)(+)/CD138(+) gating were used to measure cell markers CD138, CD38, CD56, CD117, CD3, CD13, CD33, CD19, CD7, CD20, CD22, CD34, CD28 in 47 MM patients. At the same time the morphology examination of bone marrow cells was performed. RESULTS: The suspicious myeloma cell ratio in MM patients was 9.42%-74.25% detected by flow cytometry, moreover, the myeloma cell ratio detected by morphology examination was 11.0%-80.6%, there was a good correlation between the two detection methods (r(2) = 0.54, P < 0.001). The ratio of antigen positive expression was as follows: 74.46% for CD138, 100% for CD38, 57.44% for CD56, 40.42% for CD117, 6.38% for CD13, 19.15% for CD33, 8.51% for CD20, 27.66% for CD28, 2.12% for CD22, 4.25% for CD34, 0% for CD3, 0% for CD19, 0% for CD7. CONCLUSIONS: CD45/SSC and CD38(+)/CD138(+) gating technique can accurately gate multiple myeloma cell sets which need analysis, the majority of myeloma cells expreses CD138, CD38, CD56 antigens. The immunophenotypic analysis combined with the cell morphology examination more contribute to the diagnosis and differential diagnosis of multiple myeloma.

Regulation of Mycobacterium tuberculosis D-3-phosphoglycerate dehydrogenase by phosphate-modulated quaternary structure dynamics and a potential role for polyphosphate in enzyme regulation.

D-3-phosphoglycerate dehydrogenase (PGDH) catalyzes the first reaction in the "phosphorylated" pathway of l-serine biosynthesis. In Mycobacterium tuberculosis, it is a type 1 enzyme (mtPGDH) in that it contains both an ACT domain and an ASB domain in addition to a catalytic domain. The published crystal structures (Protein Data Bank entries 1YGY and 3DC2) show a tartrate molecule interacting with cationic residues at the ASB-ACT domain interfaces and a serine molecule bound at the ACT domain interface. These sites have previously been shown to be involved in the mechanism of serine and substrate inhibition of catalytic activity. This investigation has revealed a mechanism of allosteric quaternary structure dynamics in mtPGDH that is modulated by physiologically relevant molecules, phosphate and polyphosphate. In the absence of phosphate and polyphosphate, the enzyme exists in equilibrium between an inactive dimer and an active tetramer that is insensitive to inhibition of catalytic activity by L-serine. Phosphate induces a conversion to an active tetramer and octamer that are sensitive to inhibition of catalytic activity by L-serine. Small polyphosphates (pyrophosphate and triphosphate) induce a conversion to an active dimer that is insensitive to L-serine inhibition. The difference in the tendency of each respective dimer to form a tetramer as well as slightly altered elution positions on size exclusion chromatography indicates that there is likely a conformational difference between the serine sensitive and insensitive states. This appears to constitute a unique mechanism in type 1 PGDHs that may be unique in pathogenic Mycobacterium species and may provide the organisms with a unique metabolic advantage.

Increased expression of CIP2A in cholangiocarcinoma and correlation with poor prognosis.

BACKGROUND/AIMS: To explore the expression of cancerous inhibitor of protein phosphatase 2A (CIP2A) in human cholangiocarcinoma tissues and adjacent non-cancerous normal bile duct tissues, and to investigate the clinicopathological signinficance of the CIP2A protein in cholangiocarcinoma patients. METHODOLOGY: Immunohistochemical staining was used to detect the expression of CIP2A protein in 57 cases (35 men and 22 women) of cholangiocarcinoma samples and 23 cases of para-cancerous normal bile duct samples. The results were analyzed with clinicopathological patameters and overall median survival time. RESULTS: The positive rate of CIP2A protein expression in cholangiocarcinoma tissues was significantly higher than para-cancerous normal bile duct tissues. The expression of CIP2A was found to be not correlated with age, gender, smoking, grading, staging, lymph node metastasis, tumor site and hepatitis B virus (HBV) (p>0.05). Kaplan-Meier survival analysis showed that the overall survival time in patients with positive expression of CIP2A protein were shorter than in patients with negative expression of CIP2A (long rank=5.180, p=0.023). COX regression analysis implied that expression of CIP2A protein was an independent prognostic factor for cholangiocarcinoma patients (p<0.05). CONCLUSIONS: CIP2A is overexpressed in human cholangiocarcinoma tissues, and overexpression of CIP2A correlates with poor prognosis. CIP2A expression may be a potential marker for biological malignancy.

Notch3 overexpression associates with poor prognosis in human non-small-cell lung cancer.

Notch3 receptor is one of the mammalian Notch family receptors (Notch1-4) which plays an important role in the regulation of cellular proliferation, differentiation, and apoptosis. Overexpression of Notch3 is associated with tumorigenesis. In order to assess the expression of Notch3 in Chinese non-small-cell lung cancer (NSCLC) patients and determine its association with prognosis, we designed a prospective study with five years of follow-up to evaluate Notch3 expression in NSCLC tissues and adjacent non-cancerous normal lung tissues from 131 patients undergoing surgical treatment by immunohistochemistry and western blot analysis. Notch3 had high expression in 67 of 131 cases of NSCLC (51.1 %), which was significantly higher than in adjacent noncancerous lung tissues. Moreover, Notch3 overexpression was significantly correlated with TNM stage (P = 5.41e-07 in squamous cell carcinoma, P = 5.338e-07 in adenocarcinoma) and lymph node metastasis (P = 0.00764 in squamous cell carcinoma, P = 0.01491 in adenocarcinoma). Kaplan-Meier survival analysis showed that the overall survival times in patients expressing Notch3 in NSCLC were shorter. Multivariate analysis further demonstrated that Notch3 was an independent prognostic factor for patients with NSCLC. Therefore, Notch3 might be a useful biomarker to predict the prognosis of patients with NSCLC.

CIP2A with survivin protein expressions in human non-small-cell lung cancer correlates with prognosis.

Cancerous inhibitor of protein phosphatase 2A (CIP2A) and survivin are aberrantly expressed in a wide range of human cancers, including lung tumors. In order to assess the expressions of these two proteins in Chinese non-small-cell lung cancer (NSCLC) patients and determine their correlation with prognosis, NSCLC tissues and adjacent non-cancerous normal lung tissues were collected from 97 patients undergoing surgical treatment and evaluated by immunohistochemistry staining. CIP2A or survivin immunoreactivity was detected in significantly more NSCLC tissues than in adjacent non-cancerous lung tissues (P < 0.05). Moreover, CIP2A expression in NSCLC correlated with TNM stage, while survivin expression correlated with TNM stage and lymph node metastasis. Kaplan-Meier survival analysis showed that the overall survival times in patients expressing either CIP2A or survivin protein in NSCLC were shorter. COX regression analysis indicated that expression of CIP2A protein was an independent prognostic factor for NSCLC patients (HR = 3.631, P = 0.015). Therefore, CIP2A expression in Chinese NSCLC patients may be a useful biomarker of biological malignancy.

Kinetic, mutagenic, and structural homology analysis of L-serine dehydratase from Legionella pneumophila.

A structural database search has revealed that the same fold found in the allosteric substrate binding (ASB) domain of Mycobacterium tuberculosis D-3-phosphoglycerate dehydrogenase (PGDH) is found in l-serine dehydratase from Legionella pneumophila. The M. tuberculosis PGDH ASB domain functions in the control of catalytic activity. Bacterial l-serine dehydratases are 4Fe-4S proteins that convert l-serine to pyruvate and ammonia. Sequence homology reveals two types depending on whether their α and ß domains are on the same (Type 2) or separate (Type 1) polypeptides. The α domains contain the catalytic iron-sulfur center while the ß domains do not yet have a described function, but the structural homology with PGDH suggests a regulatory role. Type 1 ß domains also contain additional sequence homologous to PGDH ACT domains. A continuous assay for l-serine dehydratase is used to demonstrate homotropic cooperativity, a broad pH range, and essential irreversibility. Product inhibition analysis reveals a Uni-Bi ordered mechanism with ammonia dissociating before pyruvate. l-Threonine is a poor substrate and l-cysteine and d-serine are competitive inhibitors with K(i) values that differ by almost 10-fold from those reported for Escherichia colil-serine dehydratase. Mutagenesis identifies the three cysteine residues at the active site that anchor the iron-sulfur complex.

The properties of tumor-initiating cells from a hepatocellular carcinoma patient's primary and recurrent tumor.

Hepatocellular carcinoma (HCC) is associated with a high morbidity and mortality due to its high rate of recurrence. However, little is known about the biological characteristics of recurrent HCC cells. A single patient's primary and recurrent HCC-derived cell lines, Hep-11 and Hep-12, respectively, were established by primary culture. These two cell lines have the same hepatitis B virus integration site and share many common amplifications and deletions, which suggest that they have the same clonal origin. While Hep-11 cells were non-tumorigenic at 16 weeks following injection of up to 10 000 cells, injection of only 100 Hep-12 cells was sufficient to initiate tumor growth, and all single Hep-12 clones were tumorigenic in immunodeficient mice. Compared with Hep-11, Hep-12 cells expressed the oval cell markers AFP, NCAM/CD56, c-kit/CD117, as well as multiple stem cell markers such as Nanog, OCT4 and SOX2. In addition, >90% of Hep-12 cells were aldehyde dehydrogenase positive. They were also less resistant to paclitaxel, but more resistant to doxorubicin, cisplatin and hydroxycamptothecin (HCPT), which had been administrated to the patient. Furthermore, Hep-12 cells expressed higher levels of poly (adenosine diphosphate-ribose) polymerase-1 (PARP-1) than Hep-11, and PARP-1 inhibition potentiated the sensitivity to HCPT in Hep-12 cells but not in Hep-11 cells. These results indicate that a large population of the recurrent HCC-derived Hep-12 cells were tumor-initiating cells and that elevated expression of PARP-1 was related to their resistance to HCPT.

[Are leukemic patient bone marrow mesenchymal stem cells malignant?].

The study was aimed to explore whether there are leukemic characteristics in the bone marrow mesenchymal stem cells (BMMSC) from leukemic patients as compared with normal controls. The mesenchymal stem cells from bone marrow of normal volunteers and patients with APL and CML were isolated, then cultured and proliferated in vitro. The morphology, growth curve and cell surface markers of two different sources mesenchymal stem cells were investigated for detecting whether the bone marrow mesenchymal stem cells derived from leukemia patients have the specific abnormal fusion gene of leukemia cells through fluorescent in situ hybridization. The results indicated that there was no significant difference between the mesenchymal stem cells derived from different subjects, the bone marrow mesenchymal stem cells derived from leukemia patients did not have the clonal malignant fusion gene as seen in the leukemia cells. Taken altogether, mesenchymal stem cells derived from leukemia patients had no biological differences as compared with those from normal volunteers, and no malignant clonal abnormality was found. It is concluded that mesenchymal stem cells derived from leukemia patients as an alternative vehicle may be used for assistant of autologous hematopoietic stem cell transplantation or cell therapy and gene therapy.

Ex-vivo analysis of CD8+ T cells infiltrating colorectal tumors identifies a major effector-memory subset with low perforin content.

Previous studies have indicated that the infiltration of CD8+ T cells in colorectal cancer is an independent predictor of increased survival but clinical observations have suggested that the cytotoxic function of CD8+ T cells infiltrating colorectal cancer may often be limited. In this study, we have assessed the phenotype of colorectal cancer CD8+ tumor-infiltrating lymphocytes (TILs) isolated ex vivo from tumor tissue, and assessed the perforin content of TIL with respect to their location using immunohistochemistry. We found that CD8+ T cells TILs isolated from colorectal cancer are mainly composed of antigen-experienced cells of effector memory type (TEM, CD45RA-CCR7-, and CD27+/CD28- or CD27-/CD28-), and contain only minor proportions of terminally differentiated CD8+ T cells (TEMRA, CD45RA+CCR7-). The perforin content of these TILs, however, is significantly lower than that of antigen-experienced T cells in PBMCs due to the much lower levels of perforin found in the CD27-CD28- subset in TILs compared with CD8+ T cells of similar phenotype in PBMCs.

[Surface morphology of CD34+ cells under atomic force microscope].

OBJECTIVE: To observe the difference in surface morphology and structure between CD34(+) cells from normal human subjects and from patients with leukemia. METHODS: Bone marrow mononuclear cells from 3 normal human subjects and 3 patients with M2b leukemia were collected by Percoll gradient centrifugation, followed by purification of CD34(+) cells by means of immunomagnetic bead separation (MiniMAC) and examination with flow cytometry. The morphology of the cells were then observed with optical and atomic force microscope (AFM) in air. RESULTS: No significant difference was identified between the two cells under optical microscope. With atomic force microscope, numerous microvilli were observed on the surface of CD34(+) cells, and the normal cells and those from the leukemic patients showed significant difference in terms of the roughness of the cell surface. CONCLUSION: Normal CD34(+) cells have more rough and erosive surface structure than leukemic CD34+ cells (M2b).

[Analysis of alkaline phosphatase staining results in 238 cases of malignant lymphoma].

OBJECTIVE: To examine the significance of alkaline phosphatase (ALP) staining in clinical characterization of malignant lymphoma cases. METHODS: Fresh peripheral blood smear obtained from 238 cases of malignant lymphoma, including 23 cases with bone marrow involvement (BMI) 189 cases without BMI(NBMI), and 26 cases of lymphoma cell leukemia (LCL), were examined by ALP staining, and the results were compared with those of 40 normal control subjects. RESULTS: With the gradual progression of lymphoma, the patients tended to have higher ALP scores, and the percentages of the subjects with a score higher than 250 in the control, NBMI, BMI and LCL were 0, 4.8%, 39.1%, 53.9% respectively, showing significant differences between the four groups. CONCLUSIONS: Increased ALP activity can be indirectly indicative of lymphoma progression, and ALP staining results may provide insight into the clinical staging and prognosis of malignant lymphoma.