Dysregulation of LncRNA ANRIL mediated by miR-411-3p inhibits the malignant proliferation and tumor stem cell like property of multiple myeloma via hypoxia-inducible factor 1α.

Long non-coding RNA (lncRNA) ANRIL has been reported to be closely related to the relapse of multiple myeloma patients. However, the functional role and underlying mechanism of lncRNA ANRIL in multiple myeloma are not known. This study aims to investigate the biological function of lncRNA ANRIL in multiple myeloma. In this study, compared with normal tissues from healthy donors, lncRNA ANRIL and HIF-1α expressions were up-regulated in tumor tissues from multiple myeloma patients. miR-411-3p expression was down-regulated in tumor tissues from multiple myeloma patients. Besides, lncRNA ANRIL can interact with miR-411-3p. HIF-1α was confirmed to be a target of miR-411-3p. Correlation analysis showed that lncRNA ANRIL expression was negatively correlated with miR-411-3p expression. HIF-1α expression was negatively correlated with miR-411-3p expression. Further transfection experiments showed that knockdown of ANRIL or overexpression of miR-411-3p significantly inhibited cell proliferation, tumor formation ability and tumor stem cell like property, promoted cell apoptosis in vitro. Finally, miR-411-3p mimic reduced tumor volume, improved survival rate, suppressed malignant proliferation and tumor stem cell like property in U266 xenograft model. Our results demonstrate that lncRNA ANRIL mediated by miR-411-3p promotes the malignant proliferation and tumor stem cell like property of multiple myeloma through regulating HIF-1α.

Phacoemulsification in the anterior chamber: An alternative surgical technique in post-vitrectomy cataract.

OBJECTIVE: To evaluate the characteristics, safety and effectiveness of a modified technique of phacoemulsification in post-vitrectomy cataracts. METHODS: This retrospective clinical trial comprised 31 patients (31 eyes) with post-vitrectomy cataract, who had undergone phacoemulsification combined with intraocular lens implantation. An alternative surgical technique known as phacoemulsification in the anterior chamber was used for nucleus management in those cases. The following parameters were evaluated: best corrected visual acuity (BCVA), ocular inflammation, intraocular pressure, endothelial cell count and surgical complications. RESULTS: Three months after surgery, the BCVA improved significantly compared with that before surgery (Z=-10.547; p<0.05). There were no significant differences in IOP before and after surgery (Z=-0.474; p>0.05). There was a statistically significant postoperative decrease in endothelial cell densities (Z=-3.916; p<0.05). The mean endothelial cell loss was -8.71%. A little inflammatory response in the anterior chamber in 11 eyes and mild corneal edema in 8 eyes were observed on the first day after surgery, which subsided after a week. The posterior capsular opacification were observed in three eyes, two of which were denser, and the YAG laser was performed for posterior capsular incision. No obvious surgical complications occurred. CONCLUSION: The modified technique of phacoemulsification, with phacoemulsification in the anterior chamber, is safe and effective to deal with post-vitrectomy cataracts.

[Effects of SUMO specific protease 1 on hPXR-mediated P-gp gene expression].

The study aimed to investigate the effects of small ubiquitin-related modifier (SUMO) specific protease 1 (SENP1) on human PXR-mediated MDR1 transcriptional activity and mRNA expression. Empty vector and expression plasmids, including PXR, SENP1 and SENP1 mutant (SENP1m) were transiently transfected into HepG2 and LS174T cells using Lipo2000. Transcriptional activity was detected by dual luciferase reporter gene assay, and mRNA level was measured using real-time polymerase chain reaction. The results showed that SENP1 could remarkably reduce the rifampicin (RIF)-induced MDR1 reporter activity and mRNA level in hPXR over expressed HepG2 and LS174T cells (P < 0.05), whereas adding SENP1m restored the RIF-induced increases (P < 0.05). These results indicated that SENP1 could repress the RIF-induced hPXR-mediated MDR1 transcriptional activity and mRNA expression.

Hydraulic properties and drought response of a tropical bamboo (Cephalostachyum pergracile).

Bamboo plants are an essential component of tropical ecosystems, yet their vulnerability to climate extremes, such as drought, is poorly understood due to limited knowledge of their hydraulic properties. Cephalostachyum pergracile, a commonly used tropical bamboo species, exhibited a substantially higher mortality rate than other co-occurring bamboos during a severe drought event in 2019, but the underlying mechanisms remain unclear. This study investigated the leaf and stem hydraulic traits related to drought responses, including leaf-stem embolism resistance (P50leaf; P50stem) estimated using optical and X-ray microtomography methods, leaf pressure-volume and water-releasing curves. Additionally, we investigated the seasonal water potentials, native embolism level (PLC) and xylem water source using stable isotope. We found that C. pergracile exhibited strong resistance to embolism, showing low P50leaf, P50stem, and turgor loss point, despite its rapid leaf water loss. Interestingly, its leaves displayed greater resistance to embolism than its stem, suggesting a lack of effective hydraulic vulnerability segmentation (HVS) to protect the stem from excessive xylem tension. During the dry season, approximately 49% of the water was absorbed from the upper 20-cm-deep soil layer. Consequently, significant diurnal variation in leaf water potentials and an increase in midday PLC from 5.87 ± 2.33% in the wet season to 12.87 ± 4.09% in the dry season were observed. In summary, this study demonstrated that the rapid leaf water loss, high reliance on surface water, and a lack of effective HVS in C. pergracile accelerated water depletion and increased xylem embolism even in the typical dry season, which may explain its high mortality rate during extreme drought events in 2019.

[Application of 3D printing modified tooth-supported cyst plug in decompression of mandibular cystic lesions].

PURPOSE: To evaluate the application value of 3D printing modified dental support cyst plug in fenestration of large jaw cystic lesions. METHODS: Forty patients with mandibular cystic disease in Xuzhou Central Hospital from October 2019 to April 2021 were selected. They were randomly divided into experimental group(3D printing group) and control group (traditional plug group), with 20 cases in each group. All enrolled patients underwent preoperative digital modeling of cystic lesions of the jaw, obtained the cystic cavity volume data of preoperative lesions, designed the windowing site according to the plan and performed decompression for jaw cysts. Three days after surgery, the patient's postoperative CBCT and Oral-scan data in the experimental group was obtained, and a digitally modified tooth-supported cyst plug with porous column channel was designed, and titanium alloy material for 3D printing was selected. In the control group, the plug was manually molded by experienced physicians. The visual analogue scale(VAS) score of pain, retention, mechanical properties of the plug and its effect on the adjacent teeth were compared between the two groups during the process of model preparation, and the changes of the cyst volume 1, 3 and 6 months after operation were compared between the two groups. SPSS 25.0 software package was used for data analysis. RESULTS: Compared with the control group, the patients in the experimental group who made titanium alloy as printing material by digital impression complained more comfortable, and the mechanical strength and stability of the cyst plug were better than those in the control group(P＜0.05). There was no significant difference in retention between the two groups(P＞0.05). The reduction rate of cyst volume in the experimental group was significantly higher than that in the traditional plug group 3 and 6 months after operation(P＜0.05). CONCLUSIONS: The modified tooth-supported titanium alloy cyst plug with digital 3D printing has good mechanical properties and stability. It has little damage to the abutment and no lateral force, and has the advantages of precision, individualization and comfort. The improved irrigation and injection channel can fully flush the cavity and speed up the narrowing of the cyst and reduce the waiting time before the second operation, which is worth promoting in clinical practice.

Effect of low-temperature stress on the survival of Meloidogyne incognita and its application in greenhouse of northern China.

We examined the effects of low temperature on egg hatching and killing rate of the 2nd instars of Meloi-dogyne incognita (J2) in the laboratory. We further evaluated the effects of two soil treatment methods on the survival rate of M. incognita in northern China in a field experiment. The results of laboratory experiment showed that survival rate of J2 was 0 after being subjected to -7 ℃ for 24 hours, and that egg hatching was completely inhibited 24 hours after being subjected to -9 ℃. The survival rate of J2 was 0 after being subjected to -1, -2, -3, and -4 ℃ for 8, 5, 3, and 1.5 d, respectively. Egg hatching was completely inhibited after being subjected to -2, -3, -4, and -5 ℃ for 9, 6, 4, and 1 d, respectively. Results of the fitting analysis showed that both the relationships between the temperature and the lethal time of J2 as well as the temperature and the non-hatching time of the eggs followed exponential functions. The results of field test showed that death rate of M. incognita in 0-50 cm soil layer after ridging treatment and 0-30 cm soil layer after leveling treatment could reach 100%, while the disease index of the former in 30-40 cm and 40-50 cm was 84.9% and 75.8%, respectively, which was lower than that in the greenhouse. Our results suggest that preventing and controlling M. incognita in greenhouses through low-tempe-rature in winter could achieve a better control effect in Yulin City and the northward region. The proposed technique is convenient and has high potential for popularization.

[Barrier effect against Bemisia tabaci and the control of tomato yellow leaf curl virus in a tomato-maize intercropping system]. / ç•ªèŒ„çŽ‰ç±³é—´å¥—ä½œå¯¹çƒŸç²‰è™±çš„å±éšœæ•ˆåº”åŠæŽ§åˆ¶ç•ªèŒ„é»„åŒ–æ›²å¶ç— æ¯’ç— çš„æ•ˆæžœ.

We examined the barrier effects against Bemisia tabaci and the potential to control the tomato yellow leaf curl virus (TYLCV) of different tomato-maize intercropping modes. Tomatoes (variety Jinshan 511) were planted at a fixed row distance either as a monoculture or together with maize (variety Xianyu 335) at the distances of 10, 20, and 30 cm. We investigated the viral index for TYLCV and the population density of B. tabaci. Compared with tomato monoculture, the natural barrier established by the maize stabilized the growth environment for tomato. When the maize spacing was 10, 20, and 30 cm, the average temperature of tomato plants from 6:00 to 20:00 decreased by 3.01, 2.26, and 1.45 ℃, the average relative humidity increased by 13.0%, 8.8%, and 6.0%, and the mean light intensity reduced by 26.1%, 20.4%, and 14.5%, respectively. The changes of those factors alleviated the adverse environmental conditions (i.e., intense light, high temperature, and lack of moisture) that promote the spread of virus-related diseases during the high-temperature period of the day, with the most effective planting distance for the maize being 10 cm. Our results suggested that the intercropping of tomato and maize had a barrier effect against B. tabaci and was able to control the TYLCV in tomato. When the planting distance for the maize was 10, 20, and 30 cm, the number of B. tabaci was 88.7%, 82.0%, and 73.9% lower than tomato monoculture, respectively. The TYLCV was inhibited, with the viral disease index being decreased by 67.3%, 59.4%, and 44.5%, respectively. Tomato-maize intercropping was also beneficial for tomato plant growth and fruit set, which could enhance tomato yield. Such effect was strengthened under higher maize density.

[Gene Mutation and Overexpression of Newly Diagnosed Multiple Myeloma Patients].

OBJECTIVE: To analyze the characteristics of gene mutation and overexpression in newly diagnosed multiple myeloma (NDMM) patients. METHODS: Bone marrow cells from 208 NDMM patients were collected and analyzed. The gene mutation of 28 genes and overexpression of 6 genes was detected by DNA sequencing. Chromosome structure abnormalities were detected by fluorescence in situ hybridization (FISH). RESULTS: Gene mutations were detected in 61 (29.33%) NDMM patients. Some mutations occurred in 5 or more cases, such as NRAS, PRDM1, FAM46C, MYC, CCND1, LTB, DIS3, KRAS, and CRBN. Overexpression of six genes (CCND1, CCND3, BCL-2, CCND2, FGFR3, and MYC) were detected in 83 (39.9%) patients, and cell cycle regulation gene was the most common. Single nucleotide polymorphisms (SNP) changes were detected in 169 (81.25%) patients, the TP53 P72R gene SNP (70.17%) was the most common. Abnormality in chromosome structure was correlated to gene overexpression. Compared to the patients with normal chromosome structure, patients with 14q32 deletion showed higher proportion of CCND1 overexpression. Similarly, patients with 13q14 deletion showed higher proportion of FGFR3 overexpression, whereas patients with 1q21 amplification showed higher proportion of CCND2, BCL-2 and FGFR3 overexpression. CONCLUSION: There are multiple gene mutations and overexpression in NDMM. However, there is no dominated single mutation or overexpression of genes. The most common gene mutations are those in the RAS/MAPK pathway and the genes of cyclin family CCND are overexpression.

Identification of prognostic splicing factors and exploration of their potential regulatory mechanisms in pancreatic adenocarcinoma.

Pancreatic adenocarcinoma (PAAD), the most common subtype of pancreatic cancer, is a highly lethal disease. In this study, we integrated the expression profiles of splicing factors (SFs) of PAAD from RNA-sequencing data to provide a comprehensive view of the clinical significance of SFs. A prognostic index (PI) based on SFs was developed using the least absolute shrinkage and selection operator (LASSO) COX analysis. The PI exhibited excellent performance in predicting the status of overall survival of PAAD patients. We also used the percent spliced in (PSI) value obtained from SpliceSeq software to quantify different types of alternative splicing (AS). The prognostic value of AS events was explored using univariate COX and LASSO COX analyses; AS-based PIs were also proposed. The integration of prognosis-associated SFs and AS events suggested the potential regulatory mechanisms of splicing processes in PAAD. This study defined the markedly clinical significance of SFs and provided novel insight into their potential regulatory mechanisms.

Diagnostic value of magnetocardiography in patients with coronary heart disease and in-stent restenosis.

BACKGROUND: In-stent restenosis (ISR) has become one of the most challenging problems in patients with coronary heart disease. At present, using non-invasive methods to assess ISR is a hot topic. In this investigation we attempted to explore the potential of magnetocardiography (MCG) in diagnosis of in-stent restenosis. METHODS: MCG was analyzed in 52 patients with coronary artery disease for three times: before stenting, one month and 7 months after successful intracoronary stenting. RESULTS: The average classification of total maps (ACTM) and the ratio of abnormal maps (RAM) were lower in 1 month after intracoronary stenting compared with that obtained before stent planting (2.91 vs 2.52, 65.74% vs 42.80%, P < 0.01), while complex ventricular excitation index (CVEI) increased from -42.63 to -20.05 (P < 0.01). In ISR subgroup (n = 16), RAM decreased in 1 month after intracoronary stenting compared to it before stenting (68.99% vs 45.26%, P < 0.05). ACTM increased in 7 months compared to that obtained in 1 month after stenting (3.15 vs 2.51, P < 0.05). According to the ROC curve, ACTM showed its unique diagnostic value in restenosis patients. The sensitivity and specificity of ACTM were 80.0%, 69.40%, respectively. Its positive predictive value and negative predictive value were 54.6% and 88.5%, respectively. CONCLUSIONS: After successful intracoronary stenting, most parameters of MCG were improved. ACTM was of prognostic value in diagnosing ISR.

The efficacy, biodistribution and safety of an inhibin DNA vaccine delivered by attenuated Salmonella choleraesuis.

DNA vaccines, the third-generation vaccines, were extensively studied. The attenuated Salmonella choleraesuis (S. choleraesuis) was widely focused as a carrier to deliver DNA vaccines in the chromosome-plasmid balanced-lethal system. The efficacy of inhibin DNA vaccine delivered by attenuated S. choleraesuis was proved in mice and cows in our previous studies. In this study, the efficacy of inhibin DNA vaccine was confirmed in rhesus monkeys. To further study the biodistribution and safety, the mice were immunized under laboratory conditions. The results of the rhesus monkeys showed the plasma IgA and IgG titres against inhibin were elevated, and the oestradiol (E2 ) and progesterone (P4 ) levels were increased with immunizing inhibin DNA vaccine. The biodistribution and safety assessment displayed the body weight, pathological change and haematology indexes where there is no significant difference between vaccinated mice and control. And the genomics analysis showed there was no integration of the inhibin gene into the mouse genome 2 months after immunization. This study indicated the inhibin DNA vaccine delivered by attenuated S. choleraesuis was safe. And this vaccine was a potential means to improve their reproductive traits in primates and other animals.

The complete mitochondrial genome of Scythropus yasumatsui (Coleoptera: Curculionidae).

The complete mitochondrial genome sequence of Scythropus yasumatsui (Coleoptera: Curculionidae) was determined by using an Illumina platform. The circular genome was 16,472 bp in length and contained 22 transfer RNA genes (tRNAs), two ribosomal RNA genes (rRNAs), 13 protein-coding genes (PCGs), and one control region. The nucleotide composition was significantly biased (A, G, C, and T was 39.74%, 10.11%, 15.41%, and 34.74%, respectively) with A + T contents of 74.49%. All PCGs were initiated with standard ATN (ATG/ATT) codons. While 10 PCGs were terminated with TAA, two PCGs were terminated with TAG (cytb and nad1), and nad5 was terminated with an incomplete stop codon TA. All tRNAs were predicted to contain typical cloverleaf secondary structures except trnS1. The phylogenetic analysis of the concatenated nucleotide sequences of 13 PCGs from 12 Curculionidae species was performed by using MrBayes 3.1.2. The results indicated that S. yasumatsui was more closely related to Naupactus xanthographus than to other species.

[Magnetocardiography changes in coronary artery disease patients with normal or unspecialized resting electrocardiogram].

OBJECTIVE: To investigate the magnetocardiography (MCG) changes in coronary artery disease (CAD) patients with normal or unspecific changes in resting electrocardiogram (ECG). METHODS: MCG mapping was performed by MCG-7 (MaGIC, Magiscan GmbH) installed in an unshielded room. All patients underwent ECG and coronary angiogram examinations and patients with normal or unspecified ECG changes and coronary artery narrowing > or = 70% in at least 1-vessel were defined as CAD group (n = 120). Patients with normal coronary angiogram served as control (n = 82). Four parameters: ACTM (average classification of total maps), RAM (ratio of abnormal maps), CVEI (complex ventricular excitation index) and R-max/T-max ratio, were analyzed in CAD and control groups. RESULTS: RAM (62% vs. 35%) and ACTM (2.62 +/- 0.98 vs. 2.29 +/- 0.90, P < 0.05) were significantly higher in CAD group than in control group. CVEI was found in abnormal zone (-100 - 0) in CAD group while in normal zone (0 - 100) in control group. The ratio of Rmax/Tmax in CAD group was also significantly higher in CAD group than in control group (6.41 +/- 3.29 vs. 4.10 +/- 2.00, P < 0.01). ROC curve analysis indicates that RAM, CVEI and Rmax/Tmax ratio were helpful parameters for CAD diagnosis and the diagnostic sensitivity was 67.1%, 65.9% and 64.3%; the specificity was 65.1%, 68.3% and 68.3% respectively. CONCLUSION: MCG was a useful tool for diagnosing chronic myocardium ischemia in CAD patients with normal or unspecific changes resting ECG.

General anesthesia versus local anesthesia for penetrating keratoplasty: a prospective study.

AIM: To examine which anesthesia general or local is more effective for penetrating keratoplasty (PKP). METHODS: Patients with indications for PKP (n=141) were enrolled in a prospective study and randomly divided into general anesthesia group (group A, 70 eyes) and local anesthesia group (group B, 71 eyes). Patients received optical PKP (group A1, 30 eyes; group B1, 30 eyes) or therapeutic PKP (group A2, 40 eyes; group B2, 41 eyes). Measurement of anterior chamber treatment time (T) for PKP patients and the ratio (R) of the area of the pupils to that of recipient graft region. T and R values, as well as perioperative and postoperative complications, were compared between groups A and B using t-test or χ (2) test. RESULTS: Patients were followed for 2wk after PKP. T was (13.45±8.64)min for group A and (7.36±5.24)min for group B, a statistically significant difference (P<0.001). The R value for group A was stable during the operation, while for PKP patients in group B the value initially increased then gradually decreased to normal after suturing. In group B, extrusion of intraocular contents occurred in 5 eyes, and iridal prolapse occured in 11 cases; no perioperative complications occurred in group A. Relapse rate for fungal keratitis was 13.04% in group B and 0% in group A. CONCLUSION: Under general anesthesia, pupils remaine stable during PKP and perioperative complications are averted. General anesthesia gives more time to treat pathological changes in the anterior chamber and treatment success rate is higher.

[Species-area and species-abundance relationships of arthropod community in various vegetation restoration areas in Zhifanggou watershed, Shaanxi province of Northwest China].

Taking the Zhifanggou watershed in Ansai, Shaanxi Province of Northwest China as a study unit, an investigation on the arthropods in 8 forest stands was conducted from 2006 to 2008, with the species-area and species-abundance relationships of the arthropods in these stands analyzed by various mathematical models. In these forest stands, the species-area relationship of the arthropods accorded with the formula S= CAm With the increase of investigation area, the species number approached to a constant, and the corresponding smallest investigation area was in the order of natural bush > natural forest > Populus davidiana+Robinia pseudoacacia forest > Hippaphae rhamnoides +Caragana mocrophylla forest> Periploca sepium forest > Hippaphae rhamnoides forest > Robinia pseudoacacia forest > Caragana mocrophylla forest, indicating that the more complex the stands, the larger the minimum area needed to be investigated. Based on sampling investigation, the species-abundance models of the arthropods in various stands were established. Lognormal distribution model (LN) was most suitable to fit the arthropod community in natural recovery stands, suggesting that in the arthropod community, there were more species with medial individual amount and fewer abundant species and rare species, and no obvious dominant species. LogCauchy distribution model (LC) was most suitable to fit the arthropod community in mixed and pure stands. As compared with natural recovery stand, mixed and pure stands had more abundant and rare species, and more dominant species.

[Aplastic anemia with macrocytic anemia: a study based on long-term follow-up].

OBJECTIVE: To elucidate the clinical features, response rate, prognosis and clonal evolution of aplastic anemia (AA) with macrocytic anemia (mAA). METHODS: The clinical features at initial diagnosis and data in follow up of mAA hospitalized from January 2000 to October 2011 were analyzed retrospectively. RESULTS: (1) Of 153/568 (26.9%) cases of mAA at initial diagnosis, 114(74.5%)were non-severe AA (NSAA), 39(25.5%)severe AA (SAA) and 0 very severe AA (VSAA), while the proportion was 16.2%, 45.2%, and 38.6% in 376 normocytic anemia AA (nAA), and the difference is statistically significant(χ(2) = 181.390; P = 0.000). The median age of mAA was significantly higher than that of nAA \[30(4 - 70)years vs 19 (3 - 68) years, P = 0.001\]. (2) There were no statistical difference in hemoglobin, absolute neutrophil count (ANC), platelet count (PLT), response rate after 6 months treatment and overall survival (OS) between mAA and nAA grouped in SAA and NSAA respectively. In SAA, the reticulocyte count (Ret) of mAA was significantly higher than that of nAA \[23.90(2.99 - 61.00)×10(9)/L vs 13.1(0 - 70.60)×10(9)/L, P = 0.000\] and the proportion of erythroid cells in bone marrow of mAA was also higher \[23.5 (0 - 58) vs 14.5 (0 - 65), P = 0.043\], while they did not differ significantly in NSAA. (3) The proportion of AA with PNH clones or abnormal cytogenetics did not differ significantly in mAA and nAA groups before treatment. The incidences of AA evolved to PNH in mAA and nAA was not statistically significant (7/153 vs 9/376, χ(2) = 1.099, P = 0.294) and so was the incidence of evolution to MDS/AML(3/153 vs 13/376, χ(2) = 0.399, P = 0.528). CONCLUSION: In presented with macrocytic anemia at initial diagnosis of AA, higher proportion of NSAA, elderly age, higher Ret and proportion of erythroid cells are features, but being no statistical difference in the response rate, OS, and proportion of clonal evolution.

[The clinical significance of evolution of paroxysmal nocturnal haemoglobinuria clones in aplastic anemia patients].

OBJECTIVE: To explore the clinical significance of evolution of paroxysmal nocturnal hemoglobinuria (PNH) clones in aplastic anemia (AA) patients. METHODS: The positive rate of PNH clones in 678 AA cases at first diagnosis from January 2002 to December 2009 were analyzed, and to compare the response rate and overall survival (OS) between AA patients with or without PNH clones. All patients were sequentially followed-up to assess the incidence rate and risk factors for AA evolving to overt PNH. RESULTS: (1) Of 119/678 (17.6%) AA patients at initial diagnosis presented with PNH clones,the positive rates of PNH clones among non-severe AA (NSAA), severe AA (SAA) and very severe AA (VSAA) were 16.7% (37/ 222), 17.3% (45/260) and 18.9% (37/196), respectively. There was no statistical difference among the three groups. (Chi2 = 0.369; P = 0.832); (2) 678 newly diagnosed AA cases were divided into 5 subgroups according to PNH clones, severity of disease and treatment regimens. There was no statistical difference among the five subgroups regarding 6m-response rate (RR) and OS. (3) Serial follow-up revealed that persistent PNH negative clones were found in 516 (76.1%) cases, and evolved to PNH positive clones after therapy in 43 (6.3%) cases. Persistent PNH positive clones were found in 72 (10.6%) cases, and disappeared the clones after treatment in 47 (6.9%) cases. There was no statistical difference among the four subgroups in terms of the 6m RR (Chi2 = 2.489,P = 0.426) and OS (P = 0.477); (4) 17 out of 678 AA cases (2.5%) evolved to overt PNH and the estimated incidence of evolution to overt PNH was (3.7 +/- 0.9)% at 10 years. The incidences of AA patients with or without PNH clones at initial diagnosis evolved to overt PNH were 3.4% and 2.3%, respectively. There was no statistical difference between the two groups, (Chi2 = 0.111; P = 0.739); and so was found in OS by Kaplan-Meier analysis (P = 0.868). Cox regression model analysis showed that none of the severity of AA, with or without PNH clone at initial diagnosis, treatment regimen and 6m RR was the risk factor for evolution to overt PNH. CONCLUSION: There is no difference between AA patients presented with or without PNH clones at initial diagnosis regarding the RR and prognosis. The appearance of PNH clones in AA is not identified as a risk factor for developing into overt PNH.

Enantioselective and regioselective organocatalytic conjugate addition of malonates to nitroenynes.

The first catalytic asymmetric conjugate addition of 1,3-dicarbonyl compounds to nitroenynes catalyzed by cinchona alkaloid-based thiourea organocatalysts has been developed. The 1,4-addition adducts were obtained solely, in moderate to good yields (up to 93%) with good enantioselectivities (up to 99% ee). This protocol affords a conceptually different entry to the precursors of pharmaceutically important chiral ß-alkynyl acid derivatives and synthetically useful chiral nitroalkynes. Notably, the protocol worked well with both aryl- and alkyl-substituted alkynyl substrates.

Highly enantioselective one-pot synthesis of spirocyclopentaneoxindoles containing the oxime group by organocatalyzed Michael addition/ISOC/fragmentation sequence.

A highly diastereo- and enantioselective organocatalytic protocol for the synthesis of biologically important spirocyclopentaneoxindoles containing the oxime functional group from easily accessible 3-allyl-substituted oxindoles and nitroolefins has been developed by a one-pot Michael addition/ISOC/fragmentation sequence.

Catalytic asymmetric construction of spirocyclopentaneoxindoles by a combined Ru-catalyzed cross-metathesis/double Michael addition sequence.

Biologically important and synthetically challenging spirocyclopentaneoxindoles with four contiguous stereocenters including one spiroquaternary stereocenter have been constructed in good yields (72-87%) with excellent diastereoselectivity (16:1→30:1 dr) and enantioselectivity (93→99% ee) by a combined Ru-catalyzed cross-metathesis/organocatalyzed asymmetric double-Michael addition sequence.