[Erythrocytosis after hematopoietic stem cell transplantation: report of 3 cases and literature review].

This is a report of three cases of three male patients. One of the patients had myelodysplastic syndrome, and two had aplastic anemia; their ages were 28, 32, and 21 years old, respectively. Two patients underwent sibling allogeneic hematopoietic stem cell transplantation, and one underwent haploidentical hematopoietic stem cell transplantation. All the patients showed elevated hemoglobin and hematocrit at 6, 16, and 9 months after transplantation, with normal white blood cells and platelets and no splenomegaly. All causes of secondary polycythemia were ruled out. Bone marrow morphology showed no erythroid hyperplasia. The PCR result for BCR-ABL (P210, P230, P190, and variants) was negative, and there were no mutations at the amino acid site 617 of JAK2, exon 12 of JAK2, exon 9 of CALR, and amino acid site 515 of MPL. All three patients had hypertension. One patient was treated with amlodipine, and the other two patients were treated with angiotensin receptor blockers. The durations of erythrocytosis for these three patients were 6 years and 3 months, 4 years and 7 months, and 5 years and 3 months, respectively through December 2022. There was no tendency for spontaneous remission. Erythrocytosis after hematopoietic stem cell transplantation is a rare complication. Previous reports in the literature suggest that the mechanism of post-transplant erythrocytosis in recipients of allogeneic hematopoietic stem cell transplantation may be different from that of recipients of other transplants.

Expression of syndecan-1, PKC and VEGF in rats with acute kidney injury and correlation between syndecan-1 and renal function.

OBJECTIVE: This study was designed to investigate the expression of syndecan-1 (Sdc-1), protein kinase C (PKC) and vascular endothelial growth factor (VEGF) in rats with acute kidney injury, as well as the association between Sdc-1 and indicators [such as serum creatinine (Scr) and blood urea nitrogen (BUN)] related to renal function. MATERIALS AND METHODS: A total of 120 clean grade 2-week-old SD rats were selected and randomized into experimental group and control group (n=60). At 12 h (T1), 24 h (T2), 36 h (T3), 48 h (T4) after the model was established, 3 mL blood from abdominal aorta was taken, and Sdc-1, PKC, VEGF, serum creatinine (Scr), urea nitrogen (BUN) and other indicators were detected by Enzyme-Linked Immunosorbent Assay (ELISA). RESULTS: The expression levels of Sdc-1, PKC and VEGF in the experimental group were increasing from T1 to T4, with statistically significant difference between every two time points (p<0.05); the expression levels of Scr and BUN in the experimental group was increasing from T1 to T4, with statistically significant difference between every two time points (p<0.05). The level of Sdc-1 in the serum of rats in the experimental group was positively correlated with Scr (r=0.668, p<0.001), negatively correlated with BUN (r=0.722, p<0.001), and positively correlated with BUN (r=0.722, p<0.001); PKC level was positively correlated with Scr (r=0.589, p<0.001), BUN (r=0.788, p<0.001), and VEGF level was positively correlated with Scr (r=0.666, p<0.001), BUN (r=0.784, p<0.001). CONCLUSIONS: As the concentration of syndecan-1 increases gradually, renal dysfunction aggravates accordingly, so syndecan-1 can be used as a marker of acute kidney injury and can be used to judge the degree of kidney injury at an early stage.

Expression of miR-205 in renal cell carcinoma and its association with clinicopathological features and prognosis.

OBJECTIVE: The aim of the present study was to examine the expression of miR-205 in renal cell carcinoma (RCC) tissue and carcinoma cells; also, we aimed to determine the association of miR-205 expression with the clinicopathological features and prognosis of RCC, and to explore the mechanism of miR-205. PATIENTS AND METHODS: Carcinoma tissue and adjacent normal tissue were collected from 60 patients with RCC, and the expression of miR-205 was determined by semi-quantitative PCR, followed by correlation analysis of miR-205 with clinicopathological features and prognosis. Subsequently, the human RCC line, ACHN, was transfected with miR-205, and the effect of miR-205 overexpression on the growth of RCC was examined by MTT assay. Moreover, the effect of miR-205 on the migration of colon cancer cells was studied by transwell assay. Additionally, immunohistochemistry and Western blot were used to investigate the epithelial-mesenchymal transition in renal cancer tissue. RESULTS: The expression of miR-205 was downregulated in RCC tissue compared with adjacent non-cancerous tissue (p < 0.01). The expression of miR-205 was closely related to the infiltration and recurrence of tumors (p < 0.01), but was not correlated with a pathological grade or clinical stage (p > 0.05). We also found that overexpression of miR-205 in RCC significantly inhibited the growth of cancer cells (p < 0.01) and significantly reduced the migration ability (p < 0.01). The epithelial-mesenchymal transition occurs in RCC, and miR-205 might inhibit cell proliferation and migration by blocking the epithelial-mesenchymal transition. CONCLUSIONS: The expression of miR-205 is low in RCC, and may play an important role throughout the progression of RCC. Further study of miR-205 may promote the development of a novel therapeutic approach for the treatment of RCC.

Association between advanced oxidation protein products (AOPP) and vascular calcification in uremic patients.

OBJECTIVE: To investigate the correlation between serum advanced oxidation protein products (AOPP) and vascular calcification in uremic patients. PATIENTS AND METHODS: The general data of included subjects were collected, and the serum AOPP, intact parathyroid hormone (iPTH), creatinine (Cre), Urea, calcium (Ca), phosphorus (P), total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterin (LDL-C), hemoglobin (Hb) and albumin (ALB) were detected. Coronary artery computed tomography (CT) scan was performed and the coronary arterial calcification score (CACS) was calculated; the whole abdomen CT scan was performed and abdominal aortic calcification index (AACI) was calculated. SPSS l9.0 software was used for data analysis. RESULTS: The coronary artery CT and detection of serum indexes showed that AOPP in positive coronary arterial calcification group was significantly increased compared with that in negative coronary arterial calcification group (59.14 ± 14.57 vs. 37.59 ± 5.31) µmol/L. The whole abdomen CT and detection of serum indexes showed that AOPP in positive abdominal aortic calcification group was significantly increased compared with that in negative abdominal aortic calcification group (60.32 ± 15.43 vs. 39.57 ± 6.25) µmol/L. AOPP in severe calcification group was significantly higher than negative group (70.72 ± 18.18 vs. 39.57 ± 6.25) µmol/L. There were no significant differences in AOPP between hypertension and non-hypertension groups, diabetic nephropathy and non-diabetic nephropathy groups. Correlation analysis showed that AOPP of uremic patients had a significantly positive correlation with logl0[CACS+1] and had a significantly positive correlation with inferior AACI. CONCLUSIONS: AOPP in positive coronary arterial calcification group and positive abdominal aortic calcification group was higher than that in negative group and AOPP in severe calcification group was significantly higher than that in negative group. AOPP of uremic patients has a significantly positive correlation with CACS and AACI.

Relationship between the gene polymorphism of osteoprotegerin and bone mineral density in hemodialysis patients.

OBJECTIVE: To investigate the relationship between the gene polymorphism of osteoprotegerin (OPG) and bone mineral density (BMD) in hemodialysis patients. PATIENTS AND METHODS: A total of 147 patients with end-stage renal disease (ESRD) who were admitted to the Weifang People's Hospital for maintenance hemodialysis between January 2014 and December 2015 were enrolled. Peripheral blood was collected from the subjects for assay of the polymorphism of A163G and G1181C loci of OPG. The measurements of the levels of RANK, RANKL, TNF-α, IL-6, PINP, CTX-I, CTX-II and TRACP5 in the isolated serum were taken. RESULTS: For the polymorphism of A163G locus on the OPG gene, the BMDs of left femoral neck and lumbar poster anterior L1-L4 of the AA genotype were significantly higher than those of the AG and GG genotypes. There was no significant difference in comparison of BMDs at the forearm (distal 1/3) between the AA genotype and AG and GG genotypes. No significant differences were found in the comparison of BMDs at all sites between AG and GG genotypes. The serum level of RANKL of the AA genotype was significantly higher than levels of AG and GG genotypes, but the levels of RANK, TNF-α, IL-6, PINP, CTX-I, CTX-II and TRACP5 were prominently lower than those levels of AG and GG genotypes. For the polymorphism of G1181C locus on the OPG gene, the BMDs of left femoral neck and lumbar poster anterior L1-L4 of the CC genotype were significantly higher than the BMDs of GG and GC genotypes, There was no significant difference in the comparison of BMDs at the forearm (distal 1/3) between the CC genotype and GG and GC genotypes. No significant differences were found in the comparison of BMDs at all sites between GG and GC genotypes. The serum level of RANKL of the CC genotype was significantly higher than the level of GG and GC genotypes. However, the levels of RANK, TNF-α, IL-6, PINP, CTX-I, CTX-II and TRACP5 were prominently lower than those levels of GG and GC genotypes. CONCLUSIONS: The polymorphisms of A163G and G1181C loci on the OPG gene were correlated with the BMD of hemodialysis patients. The genotype AA of A163G and genotype CC of G1181C were identified as the protective factors for BMD.

Down-regulation of miR-664 in cervical cancer is associated with lower overall survival.

OBJECTIVE: The aim of our study was to explore the clinicopathologic and prognostic significance of microRNA-664 expression in human cervical cancer. PATIENTS AND METHODS: qRT-PCR was used to determine the expression of miR-664 in cervical cancer tissues. The relationship between miR-664 expression levels in cervical cancer tissues and clinicopathological characteristics was estimated respectively. The survival curves of the patients were determined using the Kaplan-Meier method. Univariate and multivariate Cox regression analyses were performed. RESULTS: The expression of miR-664 is downregulated in human cervical cancer tissues when compared to the corresponding noncancerous tissues (p < 0.01). Low miR-664 expression was significantly associated with lymphatic invasion (p = 0.000), distant metastasis (p < 0.000), FIGO stage (p = 0.001), and histological grade (p = 0.008). Kaplan-Meier analysis demonstrated that low levels of miR-664 expression were associated with poorer overall survival (p < 0.001). In the multivariate analysis, low miR-664 expression was an independent prognostic factor for OS (p = 0.005). CONCLUSIONS: MiR-664 may be a promising biomarker for the detection and prognosis evaluation of cervical cancer.

First Report of Neofusicoccum parvum Causing Panicle Blight and Leaf Spot on Vitis heyneana in China.

The climbing vine, Vitis heyneana Roem. & Schult, is a member of the grape family endemic to Asia. Its fruits are used in wine production, and its roots, stems, and leaves can be used in medicinal materials. This plant is grown in Southwest China, as well as in India, Bhutan, and Nepal. Mulao Autonomous County in Guangxi Province is the only artificial cultivation area in China. During the summer of 2013, a panicle blight and leaf spot were detected on V. heyneana on four farms in Mulao Autonomous County. The symptoms were observed from the onset of florescence through fruit harvest. Brown lesions initially appeared at the base of a panicle and then extended to the whole panicle, finally causing the panicle to die and fruit to drop. When the disease developed on leaves, the symptom initially appeared as small dark brown circular spots, later enlarging into irregular spots (average diameter 6 mm) with a light brown center and dark brown rim. With severe disease, some individual leaves were affected by numerous spots, leading to premature senescence. Small sections of diseased tissue excised from 10 panicle and 10 leaf samples were plated on potato dextrose agar (PDA) and incubated at 28°C. Fungal colonies developed, initially with abundant white aerial mycelium, which turned olivaceous gray after 5 days and formed black pycnidia after 25 days. The conidia were hyaline, ellipsoidal to fusiform, externally smooth, thin-walled, and nonseptate. Thirty conidia were measured; the dimensions were 12.0 to 17.5 × 4.0 to 6.0 µm. Morphological characteristics of the isolates were similar to the descriptions of Neofusicoccum parvum (3). The isolate MPT-1 was selected as a representative for molecular identification. Genomic DNA was extracted and used for PCR to amplify the internal transcribed spacer (ITS) region and partial translation elongation factor 1-alpha (EF1-α) gene, using primers ITS1/ITS4 and EF1-728F/EF1-986R, respectively. The obtained ITS sequence (GenBank Accession No. KJ599627) and EF1-α sequence (KM921768) showed >99% homology with several GenBank sequences of N. parvum. Morphological and molecular results confirmed the isolate as N. parvum. For pathogenicity tests, detached, young healthy panicles and leaves of V. heyneana were surface-sterilized, wounded by sterile needle, and inoculated with mycelial plugs (3 mm in diameter) of four N. parvum isolates. Ten panicles and 10 leaves were used for every isolate. Control panicles and leaves were treated with sterile PDA plugs. All the samples were placed in a humid chamber (RH 90%, 28°C, 12 h of light) for 3 days. Symptoms similar to those observed in the field developed on all panicles and leaves inoculated with N. parvum isolates. N. parvum was reisolated from all inoculated, symptomatic tissues. The controls remained symptomless. N. parvum has been reported to cause trunk canker on V. vinifera (2), dieback on Cupressus funebris (3), and a leaf spot on Myristica fragrans (1). To our knowledge, this is the first report of N. parvum causing panicle blight and leaf spot on V. heyneana in China. Panicle blight caused a large number of fruits to drop and reduced the yield seriously. Some effective measures should be taken to control this disease. References: (1) V. Jayakumar et al. New Dis. Rep. 23:19, 2011. (2) J. Kaliternam et al. Plant Dis. 97:1656, 2013. (3) S. B. Li et al. Plant Dis. 94:641, 2010.

Regulatory T cells in chronic hepatitis B patients affect the immunopathogenesis of hepatocellular carcinoma by suppressing the anti-tumour immune responses.

Chronic hepatitis B (CHB) virus hepatitis B virus (HBV) infection is the key cause of hepatocellular carcinoma (HCC) in Asians. Recent studies have shown that levels of CD4(+)CD25(+) regulatory T cells (Tregs) were increased and were linked to an impaired immune response in patients with CHB. Evaluating whether Tregs are involved in the progression of CHB to HCC will provide insight into the immunopathogenesis of HCC. In the present study, we showed that circulating and liver-residing Tregs increased in CHB (n = 15) and HCC (n = 49) patients, particularly in the peripheral blood of HCC patients with HBV infection (n = 29). The increased Tregs in CHB patients suppressed the specific immune response induced by not only HBV antigen, but also by HCC tumour antigen. When peripheral blood mononuclear cells (PBMC) were co-cultured with human hepatoma cell lines that are stably transfected with HBV (HepG2.2.15), CD4(+)CD25(+) Treg populations increased and upregulated the expression of forkhead box P3 transcriptional regulator (FoxP3), cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) and glucocorticoid-induced tumour necrosis factor (TNF) receptor family gene (GITR). In contrast, PBMCs co-cultured with HepG2 cells (the parental cell line of HepG2.2.15) did not. CD4(+)CD25(+) Tregs isolated from PBMCs that were co-cultured with HepG2.2.15 cells also had a greater suppressive ability with respect to the tumour antigen-specific immune response induced by NY-ESO-1 or MAGE-A3 compared with CD4(+)CD25(+) Tregs isolated from PBMCs co-cultured with HepG2 cells. The results offer evidence that the expansion of CD4(+)CD25(+) Tregs and the enhancement of the suppressor function of CD4(+)CD25(+) Tregs induced by HBV infection-related factors could suppress the anti-tumour immune response to HCC tumour antigen and inhibit tumour immuno-surveillance against HCC, which may be involved in the immunopathogenesis from CHB to HCC.

Preparation, characterization, and anti-tumor property of podophyllotoxin-loaded solid lipid nanoparticles.

In an effort to develop an alternative formulation of podophyllotoxin suitable for drug release and delivery, podophyllotoxin-loaded solid lipid nanoparticles (PPT-SLNs) were constructed, characterized and examined for in vitro cytotoxicity and tumor inhibition. The SLNs were prepared by using a solvent emulsification-evaporation method, and their size was around 50 nm. TEM detection showed that the SLNs were homogeneous and spherical in shape, and differential scanning calorimetry (DSC) measurement revealed a new conformation of PPT-SLNs. An in vitro drug release study showed that PPT was released from the SLNs in a slow but time-dependent manner. Furthermore, the treatment of 293T and HeLa cells with PPT-SLNs demonstrated that PPT-SLNs were less toxic to normal cells and more effective in anti-tumor potency compared with unconjugated PPT. A colony forming efficiency assay showed an effective long-term cancer growth suppression of PPT-SLNs; in addition, they can also enhance the apoptotic and cellular uptake processes on tumor cells compared with PPT. These results collectively demonstrated that this SLN formulation has a potential application as an alternative delivery system for anti-tumor drugs.

Contribution of calcium and other dietary components to global variations in bone mineral density in young adults.

A research project on comparative international studies of osteoporosis using isotope techniques was organized by the International Atomic Energy Agency (IAEA) with the participation of 12 countries (Brazil, Canada, Chile, China, Croatia, Hungary, Philippines, Russia, Singapore, South Africa, Turkey, and the United Kingdom). Participating centers in 11 countries (all but the UK) made measurements and collected data on men and women aged 15 to 49 years. In addition to studies of bone mineral density (BMD) at the femoral neck and lumbar spine using DEXA, anthropometric, lifestyle, and nutritional data were also collected. The results of the nutritional studies are reviewed in this paper. Overall, about 8% of the observed variability in spine BMD could be attributed to nutritional factors in men and women; in men, no such relationship could be determined. No single nutritional component (not even calcium) stood out as being of particular importance across all participating centers.

Synergistic effect of hepatitis B virus and aflatoxin B1 in hepatocarcinogenesis in tree shrews.

An animal experiment with tree shrews was performed to detect the synergistic effects of hepatitis B virus (HBV) infection and dietary aflatoxin B1 (AFB1) in hepatocarcinogenesis. Adult healthy tree shrews (Tupaia belangeri chinensis) were divided into four groups: Group A (HBV + AFB1)--animals were infected with human HBV serum at first, then fed AFB1 diluted with milk, 150 ug/kg.bw/day, 6 days/week for 105 weeks. Group B (HBV)--animals were infected with human HBV as Group A, but no AFB1 treatment. Group C (AFB1)--animals were treated with AFB1 as Group A but no HBV infection. Group D--animals were treated neither with human HBV nor AFB1. During the experiment, blood samples and liver biopsies were taken regularly from all animals in each group. All the animals were sacrificed on the 160th week when the experiment ended. The samples of sera and liver tissues were checked for HBV markers and histological changes. Hepatocellular carcinomas (HCCs) were found only in Group A and Group C, with incidences of 67% and 30% respectively. The average time for HCC occurrence in Group A and Group C was 120.3 +/- 16.6 and 153.3 +/- 5.8 weeks respectively (P < 0.01). Even though no HCC occurred in Group B, 1 animal which died before the end of the experiment showed two large hepatocellular nodules. These results showed that there is synergistic effect between HBV and AFB1 in tree shrews' hepatocarcinogenesis, even though the hepatocarcinogenic effect played by HBV alone is rather weak.

Reliability of a short-term test for hepatocarcinogenesis induced by aflatoxin B1.

The reliability of a short-term test for hepatocarcinogenesis induced by aflatoxin B1 (AFB1) was tested by comparing the early appearance of gamma-glutamyl transpeptidase (GGT)-positive foci with the occurrence of primary liver cancer at a later stage. All rats received a basic short-term treatment with AFB1 intraperitoneally, during which three experimental groups received Chinese green tea or 2000 or 5000 ppm butylated hydroxyanisole in the diet and a control group received basic diet. Some of the rats in each group were sacrificed at the end of the short-term procedure, and the remainder were observed up to 92 weeks. The livers of all animals were examined for GGT-positive foci or primary liver tumours. The GGT-positive foci were most numerous and largest and the incidence of liver tumours was highest in the control group. These findings suggest that GGT-positive foci are a valuable preneoplastic marker for AFB1-induced hepatocarcinogenesis, that the short-term model is fairly reliable, and that both Chinese green tea and butylated hydroxyanisole inhibit AFB1-induced hepatocarcinogenesis.

[Effect of six edible plants on the development of AFB1-induced gamma-glutamyltranspeptidase-positive hepatocyte foci in rats].

Six edible plants, green tea (GT), black tea (BT), Lentinus edodes (berk) Sing (LE), Hericium erinaceus (Bull. ex Fr.) Pers. (HE), Mixture of Ganoderma Lucidum (Ley ss ex Fr.) Karst et Ganoderma Japanium (Fr.) Lloyd (MGLJ) and mung bean (MB), were tested for the effect on the development of AFB1-induced gamma-glutamyltranspeptidase positive hepatocyte foci (gamma-GT foci) using an in vivo short-term test model in rats. The rats received intraperitoneally 12 doses of initiator AFB1, 400 micrograms/kg per dose for 2 successive weeks. Two weeks after the initiation, the rats were submitted to a modified "Solt-Farber promotion program", i.e., a two weeks' feeding of a diet containing 0.015% acetylaminofluorene plus a two-third partial hepatectomy (PH) on day 7. The rats were sacrificed 10 days after PH and the livers were processed to gamma-glutamyltranspeptidase staining. The tested substances were powdered and mixed with the basal diet at the concentration level of 30% for MB and 5% for the others. The rats were fed with the diet-containing tested substances from 10 days before the AFB1 initiation to 3 days after the AFB1 conclusion. Consequently, the liver of the rats which had consumed GT showed significantly less and smaller gamma-GT foci, and those which had consumed BT, HE and LE showed somewhat less and significantly smaller foci than the control groups. It is indicated that the four diets have an inhibiting effect on AFB1-induced gamma-GT foci in different degrees. MB and MGLJ show no significant influence on the foci.