[Clinical Analysis of PEG-rhG-CSF in Mobilization of Autologous Hematopoietic Stem Cells in Hematological Tumors].

OBJECTIVE: To investigate the efficiency and optimal time of stem cell apheresis mobilized by pegylated recombinant human granulocyte colony stimulating factor (PEG-rhG-CSF) in autologous stem cell transplantation (ASCT) for hematological malignancies without monitoring pre-collection CD34+ cells. METHODS: Forty-six patients underwent stem cell mobilization were retrospectively analyzed between August 2017 and January 2022 at the First Affiliated Hospital of Fujian Medical University. 27 patients using high dose chemotherapy combined with PEG-rhG-CSF mobilization were enrolled in the PEG-rhG-CSF group, and other 19 patients mobilized with recombinant human granulocyte colony stimulating factor (G-CSF) were enrolled in G-CSF group. The mobilization and collection effects of the patients in two groups were compared. RESULTS: A total of 46 patients underwent 86 apheresis procedures, the median amount of mononuclear cell (MNC) in the PEG-rhG-CSF group and G-CSF group was 6.54（3.85-12.61）×108/kg and 6.15（1.13-11.58）×108/kg, respectively (P >0.05), the total CD34+ cells of the grafts were 11.44(1.33-65.02)×106/kg and 4.95(0.30-24.02)×106/kg (P < 0.05), with harvest timing of 14(10-20) days and 14(4-22) days, respectively (P >0.05). In the PEG-rhG-CSF group, there was a significant difference between the number of CD34+ cells collected when white blood cells (WBC) ≥10×109/L and WBC<10×109 /L, 19.04(2.85-65.02)×106/kg and 6.22(0.81-34.86)×106/kg, respectively (P < 0.05). CONCLUSION: Stem cells mobilization with PEG-rhG-CSF was highly efficient with a median mobilization time of 14 days. In the absence of peripheral blood CD34 monitoring, peripheral blood WBC≥10×109/L can be considered as a threshold for a single stem cell apheresis to collect sufficient stem cells.

An Extremely Rare Epithelioid Sarcoma Arising from the Infratemporal Fossa: A Case Report and Literature Review.

INTRODUCTION: Epithelioid sarcoma (ES) is a rare histopathological type of soft tissue sarcoma. They are subcutaneous soft tissue masses that tend to arise in extremity sites (the classic type, formerly referred to as distal type) or proximal midline region of the body (the proximal type), such as the perineum, genital tract, and pelvis. The head and neck regions are rarely affected by ES, but the infratemporal fossa (ITF) is extremely rare. ES involving the ITF has not been reported before in literature. In this paper, the imaging features of ES were reviewed in detail, and the anatomical structure and epidemiology of ITF were briefly introduced. SOURCES: We performed a systematic search from 3 databases, CNKI(China National Knowledge Internet), FMRS(Foreign Medical Literature Retrieval Service, Shenzhen METSTR Technology CO., Led. China), and PubMed, to obtain literature from January 1970 to July 2020. Epithelioid sarcoma, head and neck regions, infratemporal fossa, diagnostic imaging, anatomy, MRI, and CT were used as keywords for advanced retrieval. A study had to be found eligible for inclusion to be closely related to ES and/or ITF. A total of 129 related pieces of literature were retrieved successfully, of which 37 were closely related to this study. The case report in this article is from the Affiliated Hospital of North Sichuan Medical College. The present study was approved by the institutional review board of the Affiliated Hospital of North Sichuan Medical College and a written informed consent for the publication of the case was obtained from the patient. CASE PRESENTATION: A 33-year-old man accidentally found a slowly growing, painless mass in the left cheek more than a month ago. On plain computed tomography (CT) scan of the outpatient department, an oval slightly low-density mass with well-defined and uniform boundary in the infratemporal fossa was revealed, and on the contrast-enhanced scan, it was homogeneous and moderately enhanced. Initially, the CT appearance favored benign lesions. However, the possibility of a malignant tumor can not be excluded completely. Finally, the patient was referred to the inpatient department a tumor arose from IF and underwent a selective operation. The tumor was completely removed. Histologic findings were compatible with epithelioid sarcoma. Post-operatively, the patient received 2 cycles of radiotherapy, and there was no evidence of recurrence after 6 months follow- up. CONCLUSION: The imaging manifestations of ES are various. The anatomic structure of ITF is complex and the pathological types are various. It should be very careful in the qualitative diagnosis of tumors from the ITF, and advanced imaging techniques will be useful in imaging diagnosis.

Multilocus phylogeny and cryptic diversity of white-toothed shrews (Mammalia, Eulipotyphla, Crocidura) in China.

BACKGROUND: Crocidura, the most speciose mammalian genus, occurs across much of Asia, Europe and Africa. The taxonomy of Chinese representatives has been studied primarily based on cursory morphological comparisons and their molecular phylogenetic analyses remain unexplored. In order to understand the phylogeny of this group in China, we estimated the first multilocus phylogeny and conducted species delimitation, including taxon sampling throughout their distribution range. RESULTS: We obtained one mitochondrial gene (cytb) (~ 1, 134 bp) and three nuclear genes (ApoB, BRCA1, RAG1) (~ 2, 170 bp) for 132 samples from 57 localities. Molecular analyses identified at least 14 putative species that occur within two major well-supported groups in China. Polyphyletic C. wuchihensis appears to be composed of two putative species. Two subspecies, C. rapax rapax and C. rapax kurodai should be elevated to full species status. A phylogenetic tree based on mitochondrial gene from Asian Crocidura species showed that the C. rapax rapax is embedded within C. attenuata, making the latter a paraphyletic group. Three strongly supported undescribed species (C. sp.1, C. sp.2 and C. sp.3) are revealed from Zada County of Tibet (Western China), Hongjiang County of Hunan Province (Central China) and Dongyang County of Zhejiang Province (Eastern China), Motuo County of Tibet, respectively. The divergence time estimation suggested that China's Crocidura species began to diversify during the late Pliocene (3.66 Ma) and the Early Pleistocene (2.29 Ma), followed by a series of diversifications through the Pleistocene. CONCLUSIONS: The cryptic diversity found in this study indicated that the number of species is strongly underestimated under the current taxonomy. We propose that the three undescribed species should be evaluated using extensive taxon sampling and comprehensive morphological and morphometric approaches. Climate change since the late Pliocene and the uplift of the Qinghai-Tibet Plateau may result in the diversification and speciation of China's Crocidura species. In short, the underestimated diversity underlines the need for a taxonomic revision of Chinese Crocidura species.

Clinical Significance of MRI and Pathological Features of Giant Cell Tumor of Bone Boundary.

OBJECTIVE: To find new clues to reduce postoperative recurrence after intralesional curettage by studying MRI and pathological features of giant tumor of bone (GCTB) boundaries. METHODS: A retrospective study was performed in the departments of orthopaedic surgery and medical imaging at our hospitals from January 2006 to August 2016. A total of 16 GCTB patients confirmed by pathology were asked to participate in the present study. The age range was from 18 to 44 years (9 women and 7 men). All patients underwent MRI examination. All patients underwent en bloc resection and complete postoperative tumor segments were obtained. Five specimens were obtained randomly at the place of the segments where the GCTB boundary showed different types on MRI. Ordinary HE staining was used for all specimens and we measured the depth of local tumor cell infiltration (240 measurements). Results were expressed as means ± standard deviation. Statistical analyses were carried out with one-way ANOVA and the Student-Newman-Keuls test. P < 0.05 was considered statistically significant. The kappa test was used to analyze the degree of agreement of observers. RESULTS: A total of 16 patients (median age 30.56 years; range, 18-44 years) with GCTB (the number of distal femurs and proximal tibias was 9 and 7, respectively) were tested. The boundaries of all GCTB cases were composed of clear boundary, relatively clear boundary, and blurred boundary in different proportions on MRI. Based on continuous observation of all MRI, all boundaries were incomplete. The kappa value between two radiologists and two pathologists was 0.91 and 0.88, respectively. The average depth of local tumor cell infiltration in the clear boundary, relatively clear boundary, and blurred boundary groups was 0.42 ± 0.11 mm, 2.85 ± 0.21 mm, and 4.83 ± 0.12 mm, respectively. There was statistical difference among the three groups (F = 17.62, P < 0.05). There was also statistical difference between each of the two groups (q-value was 8.95, 14.28, and 5.21, respectively, P < 0.05). The depth of local tumor cell infiltration with blurred boundaries on MRI was the largest and the depth with clear boundaries was the smallest. CONCLUSION: The intralesional curettage boundaries need to be expanded on the basis of different types of boundaries provided by MRI.

Transgenic Expression and Identification of Recombinant Human Proinsulin in Peanut

The increased incidence of diabetes, coupled with the introduction of alternative insulin delivery methods that rely on higher doses, is expected to result in a substantial escalation in the future demand for affordable insulin. Plant-based systems offer a safe and economical method for producing pharmaceutical proteins. We used peanut (Arachis hypogaea L.) as bio-reactors to express biosafe, stable proinsulin. We designed two proinsulin analogues (FAIA and LAIA) with substitutions in their amino acid sequences. The fast-acting insulin analogue (FAIA) contains a Gly inserted between Cys19 and Gly20, as well as a Pro28Asp substitution, in the B chain. The long-acting insulin analogue (LAIA) contains a Gly inserted between Cys19 and Gly20 and two Arg residues inserted into the terminus of the B chain, as well as an Asn21Gly substitution in the A chain. Four plasmids were constructed: pROKII-Flag-FAIA, pROKII-Flag-LAIA, pCAMBIA2301-Oleosin-FAIA and pCAMBIA2301-Oleosin-LAIA. These plasmids were transferred into peanut to produce recombinant proinsulin. Western blot and GUS staining analysis indicated that some transgenic peanut successfully expressed exogenous proinsulin. Peanut seeds can act as insulin storage sites, which is the foundation for further production of recombinant proinsulin from peanut seeds.

Dihydrotestosterone regulating apolipoprotein M expression mediates via protein kinase C in HepG2 cells.

BACKGROUND: Administration of androgens decreases plasma concentrations of high-density lipid cholesterol (HDL-C). However, the mechanisms by which androgens mediate lipid metabolism remain unknown. This present study used HepG2 cell cultures and ovariectomized C57BL/6 J mice to determine whether apolipoprotein M (ApoM), a constituent of HDL, was affected by dihydrotestosterone (DHT). METHODS: HepG2 cells were cultured in the presence of either DHT, agonist of protein kinase C (PKC), phorbol-12-myristate-13-acetate (PMA), blocker of androgen receptor flutamide together with different concentrations of DHT, or DHT together with staurosporine at different concentrations for 24 hrs. Ovariectomized C57BL/6 J mice were treated with DHT or vehicle for 7d or 14d and the levels of plasma ApoM and livers ApoM mRNA were measured. The mRNA levels of ApoM, ApoAI were determined by real-time RT-PCR. ApoM and ApoAI were determined by western blotting analysis. RESULTS: Addition of DHT to cell culture medium selectively down-regulated ApoM mRNA expression and ApoM secretion in a dose-dependent manner. At 10 nM DHT, the ApoM mRNA levels were about 20% lower than in untreated cells and about 40% lower at 1000 nM DHT than in the control cells. The secretion of ApoM into the medium was reduced to a similar extent. The inhibitory effect of DHT on ApoM secretion was not blocked by the classical androgen receptor blocker flutamide but by an antagonist of PKC, Staurosporine. Agonist of PKC, PMA, also reduced ApoM. At 0.5 µM PMA, the ApoM mRNA levels and the secretion of ApoM into the medium were about 30% lower than in the control cells. The mRNA expression levels and secretion of another HDL-associated apolipoprotein AI (ApoAI) were not affected by DHT. The levels of plasma ApoM and liver ApoM mRNA of DHT-treated C57BL/6 J mice were lower than those of vehicle-treated mice. CONCLUSIONS: DHT directly and selectively down-regulated the level of ApoM mRNA and the secretion of ApoM by protein kinase C but independently of the classical androgen receptor.

Epigallocatechin gallate inhibits HBV DNA synthesis in a viral replication - inducible cell line.

AIM: To analyze the antiviral mechanism of Epigallocatechin gallate (EGCG) against hepatitis B virus (HBV) replication. METHODS: In this research, the HBV-replicating cell line HepG2.117 was used to investigate the antiviral mechanism of EGCG. Cytotoxicity of EGCG was analyzed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Hepatitis B virus e antigen (HBeAg) and hepatitis B virus surface antigen (HBsAg) in the supernatant were detected by enzyme-linked immunosorbent assay. Precore mRNA and pregenomic RNA (pgRNA) levels were determined by semi-quantitative reverse transcription polymerase chain reaction (PCR) assay. The effect of EGCG on HBV core promoter activity was measured by dual luciferase reporter assay. HBV covalently closed circular DNA and replicative intermediates of DNA were quantified by real-time PCR assay. RESULTS: When HepG2.117 cells were grown in the presence of EGCG, the expression of HBeAg was suppressed, however, the expression of HBsAg was not affected. HBV precore mRNA level was also down-regulated by EGCG, while the transcription of precore mRNA was not impaired. The synthesis of both HBV covalently closed circular DNA and replicative intermediates of DNA were reduced by EGCG treatment to a similar extent, however, HBV pgRNA transcripted from chromosome-integrated HBV genome was not affected by EGCG treatment, indicating that EGCG targets only replicative intermediates of DNA synthesis. CONCLUSION: In HepG2.117 cells, EGCG inhibits HBV replication by impairing HBV replicative intermediates of DNA synthesis and such inhibition results in reduced production of HBV covalently closed circular DNA.

A novel Locally Linear Embedding and Wavelet Transform based encoding method for prediction of MHC-II binding affinity.

The binding between peptides and MHC molecules is an important event to the cellular immunity against pathogens. The binding peptides are recognized as the epitopes, which are useful for the epitope-based vaccine design. Accurate prediction of the MHC-II binding peptides has long been a challenge in bioinformatics. Recently, most researchers are interested in predicting the binding affinity instead of categorizing peptides as "binders" or "non-binders". In this paper, we introduced a novel encoding scheme based on Locally Linear Embedding (LLE) and Wavelet Transform (WT), in which important amino acid properties were firstly selected from all properties (described in AAindex database) by using LLE, and then amino acids of peptides were replaced with these novel properties. Further, WT was adopted to extract the frequency attributes of the numerical sequences; thereby the peptides were transformed into homogeneous-length vectors. Finally, Support Vector machine Regression (SVR) was used to make quantitative prediction models based on these numerical vectors. When applied to the 16 datasets from IEDB database, our encoding scheme produced consistently better performance than other encoding schemes, indicating that our encoding scheme is an effective tool for the prediction of MHC-II binding affinity.

Preventive effect of estrogen on depression-like behavior induced by chronic restraint stress.

OBJECTIVE: To investigate the roles of estrogen and kalirin-7 in chronic restraint stress (CRS)-induced depression and the pathophysiological mechanism of depression. METHODS: Healthy female mice from Institute of Cancer Research (ICR) were randomly divided into 3 groups: control group, CRS group, and estrogen + CRS group. CRS was used to establish the animal model of depression. Forced swimming test and immunohistochemistry method were utilized to investigate the animal behavior and kalirin-7 expression in the hippocampus, respectively. RESULTS: Compared with the control group, the CRS mice displayed depression-like behaviors, including a significant reduction in body weight, a significant increase in immobility time in forced swimming test, and a dramatic decrease in kalirin-7 expression in the hippocampus. However, administration of estrogen attenuated the CRS-induced negative behaviors, and simultaneously increased kalirin-7 expression. CONCLUSION: Estrogen replacement therapy (ERT) could prevent CRS-induced depression-like behaviors in female ICR mice. Besides, kalirin-7 also plays a role in preventing CRS-induced depression-like behaviors.

[Clinical diagnosis and treatment of superior tibial fracture complicating with crotch injury of distal popliteal artery].

OBJECTIVE: To investigate the clinical effects of treatment for superior tibial fracture complicating with crotch injury of distal popliteal artery by an alternative micro-surgical operation. METHODS: During Feb. 2002 to Oct. 2007, there were 19 patients with superior tibial fracture complicating with crotch injury of distal popliteal artery included 15 males and 4 females aged from 21 to 48 years (means 35 years). There were 6 cases complicating with fracture of tibial plateau, 3 cases complicating with nerve injury. The tibial fracture were fixed with external fixator and the anterior tibial artery and vein and posterior tibial artery and vein were treated by transplantable great saphenous vein (Y-shape) combined with windowing for interosseous membrane by the posterior and anterolateral incision. Evaluations of clinical effect were performed according to Rasmussen functional score system. RESULTS: All patients survived after operation. All fracture achieved bony union, the union time was from 3 to 14 months (means 5.5 months). All patients were followed-up for from 8 to 23 months (means 13 months). The mean Rasmussen functional score was (27.0 + 2.9). The results were excellent in 11 cases, good in 7, and fair in 1. CONCLUSION: The surgical exploration should be done as soon as possible when diagnosis of injuries of large arteries is definite or highly suspected. Simultaneous reconstruction of both posterior tibial artery and anterior tibial artery associated with vein can reduce the rate of disability and recover function of limb.

HCV NS2 protein inhibits cell proliferation and induces cell cycle arrest in the S-phase in mammalian cells through down-regulation of cyclin A expression.

Chronic hepatitis C virus (HCV) infection often leads to liver cancer. NS2 protein is a HCV hydrophobic transmembrane protein that associates with several cellular proteins in mammalian cells. In this report, we investigated the functions of NS2 protein by examining its effects on cell growth and cell cycle progression. Stable NS2-expressing HeLa and Vero cell lines were established by transfection of the cells with pcDNA3.1(-)-NS2 followed by selection of the transfected cells in the presence of G418. We found that the proliferation rates of both NS2-expressing cell lines were inhibited by 40-50% compared with the control cells that were transfected with pcDNA3.1(-) control vector. Cell cycle analysis of these NS2-expressing cell lines shows that the proportion of cells in the S-phase increased significantly compared to that of control cells that do not express NS2 protein, suggesting NS2 protein induces cell cycle arrest in the S-phase. Further studies showed that the induction of cell cycle arrest in the S-phase by NS2 protein is associated with the decrease of cyclin A level. In contrast, the expression of NS2 protein does not affect the levels of cyclin-dependent kinase CDK2, CDK4, cyclin D1, or cyclin E. Our results suggest that HCV NS2 protein inhibits cell growth and induces the cell cycle arrest in the S-phase through down-regulation of cyclin A expression, which may be beneficial to HCV viral replication. Our findings not only provide information in the understanding mechanism of HCV infection, but also provide guidance for the future development of potential therapeutics for the prevention and treatment of the viral infection.

Hepatitis C virus non-structural 5A protein can enhance full-length core protein-induced nuclear factor-kappaB activation.

AIM: To study the effects of hepatitis C virus (HCV) core and non-structural 5A (NS5A) proteins on nuclear factor-kappaB (NF-kappaB) activity for understanding their biological function on chronic hepatitis caused by HCV infection. METHODS: Luciferase assay was used to measure the activity of NF-kappaB in three different cell lines cotransfected with a series of deletion mutants of core protein alone or together with NS5A protein using pNF-kappaB-Luc as a reporter plasmid. Western blot and indirect immunofluorescence assays were used to confirm the expression of proteins and to detect their subcellular localization, respectively. Furthermore, Western blot was also used to detect the expression levels of NF-kappaB/p65, NF-kappaB/p50, and inhibitor kappaB-a (IkappaB-a). RESULTS: The wild-type core protein (C191) and its mutant segments (C173 and C158) could activate NF-kappaB in Huh7 cells only and activation caused by (C191) could be enhanced by NS5A protein. Moreover, the full-length core protein and its different deletion mutants alone or together with NS5A protein did not enhance the expression level of NF-kappaB. The NF-kappaB activity was augmented due to the dissociation of NF-kappaB-IkappaB complex and the degradation of IkappaB-a. CONCLUSION: NF-kappaB is the key transcription factor that can activate many genes that are involved in the cellular immune response and inflammation. Coexpression of the full-length core protein along with NS5A can enhance the NF-kappaB activation, and this activation may play a significant role in chronic liver diseases including hepatocellular carcinoma associated with HCV infection.