Lymphocyte Antigen 6 Complex, Locus C+ Monocytes and Kupffer Cells Orchestrate Liver Immune Responses Against Hepatitis B Virus in Mice.

To understand the mechanism(s) of age-dependent outcomes of hepatitis B virus (HBV) infection in humans, we previously established an age-related HBV mouse model in which 6-week-old (N6W) C3H/HeN mice exhibited virus tolerance whereas 12-week-old (N12W) counterparts presented virus clearance. By investigating the hepatic myeloid cell dynamics in mice of these two ages, we aim to identify factors associated with HBV clearance. C3H/HeN mice were transfected with an HBV plasmid by hydrodynamic injection. Serum HBV markers were monitored weekly. Hepatic leucocyte populations and their cytokine/chemokine productions were examined at baseline, day 3 (D3), day 7 (D7), and day 14 after injection. C-C chemokine receptor type 2 (CCR2) antagonist and clodronate (CLD) were respectively administered to N12W and N6W mice to study the roles of lymphocyte antigen 6 complex, locus C (Ly6C)+ monocytes and Kupffer cells (KCs) in viral clearance. N12W mice had a significantly higher number of TNF-α-secreting Ly6C+ monocytes and fewer IL-10-secreting KCs at D3 in the liver than their younger N6W counterparts after HBV transfection. In addition, the elevated number of interferon-γ+ TNF-α+ CD8+ T cells at D7 was only seen in the older cohort. The enhanced Ly6C+ monocyte induction in N12W mice resulted from elevated C-C motif chemokine ligand 2 (CCL2) secretion by hepatocytes. CCR2 antagonist administration hampered Ly6C+ monocyte recruitment and degree of KC reduction and delayed HBV clearance in N12W animals. Depletion of KCs by CLD liposomes enhanced Ly6C+ monocyte recruitment and accelerated HBV clearance in N6W mice. Conclusions: Ly6C+ monocytes and KCs may, respectively, represent the resistance and tolerance arms of host defenses. These two cell types play an essential role in determining HBV clearance/tolerance. Manipulation of these cells is a promising avenue for immunotherapy of HBV-related liver diseases.

[Consumptions of Meat, Dietary Fat, and Fatty-acids and Prevalence of Overweight/Obesity in Children and Adolescents-a Cross-sectional Survey in Chengdu].

OBJECTIVES: To determine the prevalence of overweight and obesity in children and adolescents aged 7-15 years in Longquanyi District of Chengdu and its association with consumptions of meat, dietary fat and fatty-acids. METHODS: A total of 1 811 children and adolescents aged 7-15 years in Longquanyi District were selected using stratified cluster sampling strategy. Their body mass, height and waist circumference were measured. The prevalence of overweight/obesity was estimated based on body mass index (BMI), body mass index standard deviation score (BMI SDS), and waist-to-height ratio (WHtR). Daily consumptions of meat, dietary fat and fatty-acids were calculated using data collected through a food frequency questionnaire and 3-d 24 h dietary recall. The children with overweight/obesity were compared with those with normal body mass in food/nutrient consumptions using Wilcoxon tests. The BMI SDS, WHtR, and prevalence of overweight and obesity were also compared between those having low, moderate and high food/nutrient consumptions using Chi-square tests or Kruskal-Wallis tests. RESULTS: About 10.34% and 6.59% of participants were found to be overweight and obese, respectively. Boys had higher prevalence of overweight (12.05%) and overweight/obesity (18.97%) than girls (8.55%, 14.80%) ( P<0.05). Girls consumed more meat (including red meat and white meat), saturated fatty-acid (SFA) and monounsaturated fatty-acid (MUFA) than boys ( P<0.05). The consumptions of meat (both red meat and white meat), SFA and MUFA increased with age ( P<0.05). Overweight/obese girls consumed more SFA, MUFA and fat (%EN) than those of normal weight. The BMI SDS and WHtR of girls increased with fat (%EN) consumptions ( P<0.05). The BMI SDS of girls also increased with MUFA consumptions ( P<0.05). CONCLUSIONS: Consumptions of red meat, dietary fat, SFA, and MUFA are associated with overweight/obesity of girls aged 7-15 years in Chengdu. Further studies are needed to understand the gender differences.

Longitudinal Contraction Venoplasty in Prevention of Internal Jugular Vein Thrombosis After Free Flap Vascular Anastomosis.

PURPOSE: The vascular anastomosis of free flap with the internal jugular vein stump is susceptible to thrombosis, thus resulting in flap compromise or failure. This study aims to explore the method of longitudinal contraction venoplasty in the prevention of internal jugular vein stump thrombosis after free flap vascular anastomosis and its feasibility. PATIENTS AND METHODS: A retrospective review was performed of 16 patients who underwent internal jugular vein resection because of cancer ablation and longitudinal contraction venoplasty to prevent internal jugular vein stump thrombosis after free flap vascular anastomosis from January 2013 through December 2014 at Second Xiangya Hospital. The methods for longitudinal contraction venoplasty and its efficacy are reported. RESULTS: Venous anastomosis of all flaps was performed with the internal jugular vein stump, and single venous anastomosis was made in all cases. Postoperatively, all flaps survived completely, without vascular compromise. All the patients were followed for approximately 9 to 33 months, and they were satisfied with the esthetic and functional results of the donor and recipient sites after reconstruction. CONCLUSIONS: Longitudinal contraction venoplasty may be an effective new method for prevention of internal jugular vein thrombosis after free flap vascular anastomosis.

[Dietary Fiber and Pubertal Development among Children and Adolescents--a Cross-sectional Study in Chengdu, Sichuan].

OBJECTIVE: To determine the association between intake of dietary fiber and pubertal development among children and adolescents in Chengdu. METHODS: A cross-sectional survey was undertaken in 1 340 children and adolescents aged 9-15 years. Data about dietary intake were collected through 24-h dietary self-recall. Pubertal development was measured by trained investigators using Tanner criteria. Consumptions of total fiber and fiber from different sources were compared among the participants with different stages of pubertal development. RESULTS: Data from 1 328 children and adolescents were analyzed. Boys (n = 667) at a later stage of pubertal development consumed less total fiber and fruit fiber than those at an earlier stage (P < 0.05). Similarly, girls (n = 651) at a later stage of pubertal development consumed less fruit fiber than those at an earlier stage (P < 0.05). CONCLUSION: Dietary fiber intake, especially fruit fiber, is lower in children and adolescents with early commencement of puberty development. Further studies are needed to establish the relationship between dietary fiber and pubertal development.

In Vivo Contact Characteristics of Distal Radioulnar Joint With Malunited Distal Radius During Wrist Motion.

PURPOSE: To determine whether distal radioulnar joint (DRUJ) contact characteristics were altered in patients with malunited distal radius fractures. METHODS: We obtained computed tomography scans at 5 positions of both wrists of 6 patients who had unilateral malunited distal radius fractures with dorsal angulation from 10° to 20° and ulnar variance less than 3 mm. We reconstructed 3-dimensional images and mapped contact regions of DRUJ by calculating the shortest distance between the 2 opposing bones. The contact areas of the DRUJ were measured and the contact region centers were calculated and analyzed. The values of the malunited side were compared with those of the contralateral uninjured side. RESULTS: In the uninjured wrist, the contact areas of the DRUJ increased slightly from wrist flexion to extension and ulnar deviation. In the malunited wrist, we found the contact areas of DRUJ to be progressively reduced from 20° flexion to neutral, 40° extension, and 20° extension, to ulnar deviation. The centroid of this area on the sigmoid notch moved to distal from flexion to extension. Compared with the contralateral uninjured wrist, the contact area significantly decreased during wrist extension and ulnar deviation, and significantly increased during wrist flexion. The centroids of this area on sigmoid notch all moved volarly in all selected wrist positions. CONCLUSIONS: The contact areas of the DRUJ and the centroid of contact area on sigmoid notch are altered in patients with malunited distal radius fractures. The contact area of the DRUJ increases during wrist flexion and decreases during wrist extension and ulnar deviation. The centroids of the contact area on sigmoid notch move volarly during wrist flexion-extension and ulnar deviation. CLINICAL RELEVANCE: The in vivo findings suggest that alterations in joint mechanics may have an important role in the dysfunction associated with these injuries.

[Consumptions of Meat and Dairy Products, Zinc Intake and Pubertal Development in Adolescents in Chengdu].

OBJECTIVE: To determine the associations between meat, dairy and zinc intake and pubertal development in adolescents in Chengdu. METHODS: A total of 1320 children and adolescents aged 9-15 years in Chengdu were recruited using a stratified cluster sampling strategy. Dietary intake was assessed by the food frequency questionnaire (FFQ) and 3-day 24-hour dietary recall. Pubertal development was evaluated through physical examinations. Consumptions of meat and dairy, and zinc intake were compared between groups with different levels of pubertal development according to the Tanner criteria. RESULTS: The median age of spermarche was 13. 00 years. The boys who had had spermarche consumed more meat (including red meat) and dairy products than those who had not yet (P<0. 05). Daily consumption of total meat was positively correlated with the level of pubertal development (P<0. 05). The median age of menarche was 12. 11 years. The girls who had had menarche consumed more meat and less diiry products than those who had not yet (P<0. 05). Daily consumption of dairy products was negatively associated with breast development and the level of pubertal development (P < 0. 05). CONCLUSION: Consumptions of meat, red meat and dairy products are associated with pubertal development in adolescents in Chengdu. However, the differences between boys and girls warrant further studies.

Changes in contact site of the radiocarpal joint and lengths of the carpal ligaments in forearm rotation: an in vivo study.

PURPOSE: To examine the contact site of the radiocarpal joint and lengths of carpal ligaments at different forearm rotations in vivo. Rotation of the forearm could exert noteworthy influence on mechanics of the wrist, and understanding how forearm rotation influences wrist mechanics may help treat carpal disorders because wrist position closely relates to forearm rotation. METHODS: We obtained computed tomography scans of the wrists of 8 volunteers at the following 7 positions of forearm rotation: neutral; 30°, 60°, and 90° of pronation; and 30°, 60°, and 90° of supination. Three-dimensional images of the carpals and distal radius and ulna were reconstructed with software. Subsequently, the contact site of the scaphoid and lunate on the radial articular surface and the lengths of 8 carpal ligaments between their respective origin and insertion points were measured and compared among different positions of forearm rotation. RESULTS: We found that the contact site of the scaphoid on the distal radius moved between 0.2 and 2.0 mm during forearm rotation. The lengths of the 3 ulnar carpal ligaments (ie, ulnocapitate [UC], ulnolunate [UL], and ulnotriquetral [UT] ligaments) showed the greatest and significant change. From neutral position to pronation, the UC, UL, and UT ligaments shortened significantly. From neutral position to supination, the UT ligament lengthened significantly, but the radioscaphocapitate, UC, UL, and dorsal intercarpal ligaments decreased significantly. CONCLUSIONS: During forearm rotation, the contact site of the scaphoid and the lunate on the distal radial articular surface changed minimally. The lengths of 3 ulnar carpal ligaments (UC, UL, and UT ligaments) changed substantially. CLINICAL RELEVANCE: Our findings will help elucidate carpal biomechanics during forearm rotation. The findings may inform decisions about how to reduce the load to these carpal ligaments when treating the carpal disorders.

In vivo gliding and contact characteristics of the sigmoid notch and the ulna in forearm rotation.

PURPOSE: To investigate shifting of the contact center over the surfaces of 2 opposing bones of the distal radioulnar joint during forearm rotation. METHODS: We recruited 8 volunteers and used their right wrists. Serial computed tomography scans were obtained with the forearm at neutral position and 6 other positions of forearm rotation. We reconstructed 3-dimensional images and mapped contact regions of both the sigmoid notch and ulnar head by calculating the shortest distance between the 2 opposing bones. The center of contact was also defined and plotted against the distal radioulnar joint rotation to determine the sliding distance over the surfaces of the 2 bones. RESULTS: During forearm rotation, the maximal sliding of the sigmoid notch over the ulnar head was 7.4 mm in forearm pronation and 9.2 mm in forearm supination, which occurred in volar-dorsal direction primarily. Sliding of the ulnar head over the sigmoid notch was more limited, measuring 4.7 mm during pronation and 2.3 mm during supination. Most of the motion occurred between 30° pronation and 60° supination. In the proximal-distal direction, the contact site of the sigmoid notch with the ulnar head translated distally 1.6 mm during pronation and proximally 0.7 mm during supination. CONCLUSIONS: During forearm rotation, the sigmoid notch slides substantially against the ulnar head at each part of the forearm rotation arc. The sliding of the ulnar head over the sigmoid notch is smaller, most of which is at the range from moderate forearm pronation to slight supination. The contact site of the sigmoid notch with the ulnar head moves slightly distally during forearm pronation and proximally during supination. CLINICAL RELEVANCE: The in vivo findings provide more detailed information and insight into distal radioulnar joint motion kinematics.

IRF8 and its related molecules as potential diagnostic biomarkers or therapeutic candidates and immune cell infiltration characteristics in steroid-induced osteonecrosis of the femoral head.

PURPOSE: Steroid-induced osteonecrosis of the femoral head (SONFH) was a refractory orthopedic hip joint disease in the young and middle-aged people, but the pathogenesis of SONFH remained unclear. We aimed to identify the potential genes and screen potential therapeutic compounds for SONFH. METHODS: The microarray was obtained for blood tissue from the GEO database, and then it identifies differentially expressed genes (DEGs). The DEGs were analyzed to obtain the differences in immune cell infiltration. The gene functional enrichment analysis of SONFH was analyzed. The PPI of DEGs was identified through the STRING database, and the cluster modules and hub genes were ascertained using MCODE and CytoHubba, and the ROC curve of hub genes was analyzed, and the tissues distribution of hub genes was understood by the HPA, Bgee and BioGPS databases. The hub genes and target miRNAs and corresponding upstream lncRNAs were predicted by TargetScan, miRDB and ENCORI database. Subsequently, we used CMap, DGIdb and L1000FWD databases to identify several potential therapeutic molecular compounds for SONFH. Finally, the AutoDockTools Vina, PyMOL and Discovery Studio were employed for molecular docking analyses between compounds and hub genes. RESULTS: The microarray dataset GSE123568 was obtained related to SONFH. There were 372 DEGs including 197 upregulated genes and 175 downregulated genes by adjusted P value < 0.01 and |log2FC|> 1. Several significant GSEA enrichment analysis and biological processes and KEGG pathway associated with SONFH were identified, which were significantly related to cytoskeleton organization, nucleobase-containing compound catabolic process, NOD-like receptor signaling pathway, MAPK signaling pathway, FoxO signaling pathway, neutrophil-mediated immunity, neutrophil degranulation and neutrophil activation involved in immune response. Activated T cells CD4 memory, B cells naïve, B cells memory, T cells CD8 and T cells gamma delta might be involved in the occurrence and development of SONFH. Three cluster modules were identified in the PPI network, and eleven hub genes including FPR2, LILRB2, MNDA, CCR1, IRF8, TYROBP, TLR1, HCK, TLR8, TLR2 and CCR2 were identified by Cytohubba, which were differed in bone marrow, adipose tissue and blood, and which had good diagnostic performance in SONFH. We identified IRF8 and 10 target miRNAs that was utilized including Targetsan, miRDB and ENCORI databases and 8 corresponding upstream lncRNAs that was revealed by ENCORI database. IRF8 was detected with consistent expression by qRT-PCR. Based on the CMap, DGIdb and L1000FWD databases, the 11 small molecular compounds that were most strongly therapeutic correlated with SONFH were estradiol, genistein, domperidone, lovastatin, myricetin, fenbufen, rosiglitazone, sirolimus, phenformin, vorinostat and vinblastine. All of 11 small molecules had good binding affinity with the IRF8 in molecular docking. CONCLUSION: The occurrence of SONFH was associated with a "multi-target" and "multi-pathway" pattern, especially related to immunity, and IRF8 and its noncoding RNA were closely related to the development of SONFH. The CMap, DGIdb and L1000FWD databases could be effectively used in a systematic manner to predict potential drugs for the prevention and treatment of SONFH. However, additional clinical and experimental research is warranted.

A study of clinical anatomy of the safe zone for the volar approach for wrist arthroscopy.

OBJECTIVE: The current study was to perform an anatomical observation of the volar approach for wrist arthroscopy and to establish the safe zone for this approach. METHODS: Eight preserved specimens and 2 fresh specimens were used to simulate the medial-to-lateral operation and to mark the volar approach. Based on anatomical measurements of the volar approach, the closest distances from the 1/2, 6R, 6U, VR, VR' and VU approaches to the adjacent important structures were established. RESULTS: The closest distance from the 1/2 approach to the superficial branch of the radial nerve was 2.4±1.5mm. The closest distances from the 6U and 6R approaches to the dorsal carpal branch of the ulnar nerve were 16.2±1.3mm and 9.0±2.4mm, respectively. The closest distances from the VR and VR' approaches to the palmar cutaneous branch of the median nerve were 6.7±1.1mm and 2.8±0.9mm, respectively; the closest distances to the radial artery were 6.3±4.0mm and 10.0±3.4mm, respectively. Moreover, both of the two approaches passed through the base of the flexor carpi radialis tendon. The closest distance from the VU approach to the ulnar artery and flexor digitorum profundus tendon were 3.3±1.4mm and 0.3±2.5mm, respectively. CONCLUSIONS: In conclusion, a safe zone could be located for the establishment of the volar approach for wrist arthroscopy. LEVEL OF EVIDENCE: III; retrospective study with no control group.

In vivo changes in contact regions of the radiocarpal joint during wrist hyperextension.

PURPOSE: Distal radius and scaphoid fractures commonly occur after a fall with the hand outstretched and wrist hyperextended. We investigated contact characteristics of the radiocarpal joint in neutral position, hyperextension, and hyperextension combined with radial deviation in vivo. METHODS: Eight volunteers without a known history of wrist injury were enrolled. We obtained computed tomography scans with 3-dimensional reconstructions of the subjects' right wrists in neutral, hyperextension, and hyperextension with 10° of radial deviation. The contact regions of the radiocarpal joint were mapped. The direction and distance of changes in the contact region centers were recorded and analyzed. RESULTS: From neutral position to hyperextension, the contact of the scaphoid substantially shifted from the middle to the dorsal part of the articular surface of the radius in 5 of the 8 wrists. With these wrists further deviated radially, the contact shifted to the surface over the radial styloid. In the other wrists, the contact of the scaphoid remained in the center of the radial articular surface. In all wrists, the contact of the radius on the scaphoid shifted from the proximal lateral surface of the scaphoid to the proximal dorsal surface of the scaphoid, and the contact of the radius on the lunate shifted dorsally. CONCLUSIONS: During wrist hyperextension, the contact of the scaphoid on the distal radius exhibited 2 possible types of changes: either shifting from the mid-portion to the dorsal ridge of the articular surface of the radius or remaining at the center of the articular surface. Combined wrist hyperextension with radial deviation caused the scaphoid to contact the radius over the radial styloid. The contact of the radius on the scaphoid shifted from proximal lateral to proximal dorsal scaphoid, and that on the lunate shifted dorsally. CLINICAL RELEVANCE: This study provided in vivo mechanical findings to improve our understanding of the mechanism of hyperextension injuries of carpus.

Identification of potential autophagy-related genes in steroid-induced osteonecrosis of the femoral head via bioinformatics analysis and experimental verification.

PURPOSE: Steroid-induced osteonecrosis of the femoral head (SONFH) is a refractory orthopaedic hip joint disease that occurs in young- and middle-aged people. Previous experimental studies have shown that autophagy might be involved in the pathological process of SONFH, but the pathogenesis of autophagy in SONFH remains unclear. We aimed to identify and validate the key potential autophagy-related genes involved in SONFH to further illustrate the mechanism of autophagy in SONFH through bioinformatics analysis. METHODS: The GSE123568 mRNA expression profile dataset, including 10 non-SONFH (following steroid administration) samples and 30 SONFH samples, was downloaded from the Gene Expression Omnibus (GEO) database. Autophagy-related genes were obtained from the Human Autophagy Database (HADb). The autophagy-related genes involved in SONFH were screened by intersecting the GSE123568 dataset with the set of autophagy genes. The differentially expressed autophagy-related genes involved in SONFH were identified with R software. In addition, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of the differentially expressed autophagy-related genes involved in SONFH were conducted by using R software. Then, the correlations between the expression levels of the differentially expressed autophagy-related genes involved in SONFH were confirmed with R software. Moreover, the protein-protein interaction (PPI) network was analysed by using the Search Tool for the Retrieval of Interacting Genes (STRING), significant gene cluster modules were identified with the MCODE Cytoscape plugin, and hub genes among the differentially expressed autophagy-related genes involved in SONFH were screened by using the CytoHubba Cytoscape plugin. Finally, the expression levels of the hub genes of the differentially expressed autophagy-related genes involved in SONFH were validated in hip articular cartilage specimens from necrotic femur heads (NFHs) by using the GSE74089 dataset and further verification by qRT-PCR. RESULTS: A total of 34 differentially expressed autophagy-related genes were identified between the peripheral blood samples of SONFH patients and non-SONFH patients based on the defined criteria, including 25 upregulated genes and 9 downregulated genes. The GO and KEGG pathway enrichment analyses revealed that these 34 differentially expressed autophagy-related genes involved in SONFH were particularly enriched in death domain receptors, the FOXO signalling pathway and apoptosis. Correlation analysis revealed significant correlations among the 34 differentially expressed autophagy-related genes involved in SONFH. The PPI results demonstrated that the 34 differentially expressed autophagy-related genes interacted with each other. Ten hub genes were identified by using the MCC algorithms of CytoHubba. The GSE74089 dataset showed that TNFSF10, PTEN and CFLAR were significantly upregulated while BCL2L1 was significantly downregulated in the hip cartilage specimens, which was consistent with the GSE123568 dataset. TNFSF10, PTEN and BCL2L1 were detected with consistent expression by qRT-PCR. CONCLUSIONS: Thirty-four potential autophagy-related genes involved in SONFH were identified via bioinformatics analysis. TNFSF10, PTEN and BCL2L1 might serve as potential drug targets and biomarkers because they regulate autophagy. These results expand the autophagy-related understanding of SONFH and might be useful in the diagnosis and prognosis of SONFH.

A 14-amino acids deletion in BmShadow results to non-moult on the 2nd instar in the bivoltine silkworm, Bombyx mori.

The Bombyx mori Shadow gene (BmShadow) belongs to the superfamily of cytochrome P450 genes. To elucidate the function of the BmShadow gene and its association with diapause, we employed the CRISPR/Cas9 system to knock out the BmShadow gene in the bivoltine strain Qiufeng. The mutant (BmShadow-/-) was obtained in G2, exhibiting a 42-base deletion corresponded exactly to the amino acids regions from positions 155 to 168. The larvae of BmShadow-/- cannot moult at the pre-moulting stage of the 2nd instar. When the BmShadow-/- larvae were fed with 20E analogue at the late stage of the 2nd instar, they were rescued and developed into the 3rd instar. Rescue experiments indicated that the 20E concentration of BmShadow-/- larvae was significantly lower than that in WT larvae, and the 20E concentration of BmShadow-/- larvae which fed 20E analogue was restored to normal levels. Interestingly, the BmShadow-/- larvae could not moult on the 1st instar when they hatched from eggs after being stored at 5 °C for 40 days or after hibernation, suggesting that the 20E transported from the mother was partially consumed in the diapause maintenance phase. Our study confirmed that BmShadow is involved in 20E synthesis and a 14-amino acids region from position 155 to 168 was essential for its function, also there appears to be no other compensation pathway in vivo, which offered an important potential target locus for the control of silkworm development and the biological control of agricultural and forestry pests.

HOXC10 promotes migration and invasion via the WNT-EMT signaling pathway in oral squamous cell carcinoma.

As a master regulator of embryonic morphogenesis, homeodomain-containing gene 10 (HOXC10) has been found to promote progression of human cancers and indicate poor survival outcome. Therefore, we concentrate on elucidating the role of HOXC10 in progression of oral squamous cell carcinoma (OSCC). In our study, the expression of HOXC10 was significantly increased in human OSCC samples and was significantly correlated with TNM stage and lymph node metastasis. Upregulation of HOXC10 indicated a poor overall survival of OSCC patients according to the Kaplan-Meier survival curves. Furthermore, HOXC10-knockdown dramatically suppressed migration, invasion, and expression of N-Cadherin, Vimentin and Snail, as well as increased E-cadherin level both in vivo and in vitro. Bioinformatics and cellular study further confirmed that HOXC10 may promote invasion and migration of OSCC cells by regulating the WNT/epithelial-mesenchymal transition (EMT) signaling pathway. These findings suggest that HOXC10 plays a pivotal role in the metastasis of OSCC and highlight its usefulness as a potential prognostic marker or therapeutic target in human OSCC.

[Detection and Diagnostic Values of JAK2, CALR, MPL Gene Mutations in 208 Cases of BCR/ABL1 Negative Chronic Myeloproliferative Diseases].

OBJECTIVE: To detect the JAK2, CALR and MPL gene mutations in patients with BCR/ABL1 negative chronic myeloproliferative diseases（BCR/ABL1-CMPD）and to evaluate their diagnostic value. METHODS: Two hundred and eight cases of BCR/ABL1-CMPD comprising of 146 cases of essential thrombocythemia（ET）, 37 cases of polycythemia vera（PV）and 25 cases of primary myelofibrosis（PMF）from March 2012 to December 2015 were enrolled in the BCR/ABL1-CMPD, while 124 cases of secondary thrombocythemia and 73 cases of secondary polycythemia were enrolled in the control group. The genomic DNA and total RNA Were isolated from bone marrow or peripheral blood, then the exons 12 to 20 of JAK2 gene, exon 10 of MPL gene and exons 3 to 9 of CALR gene were analyzed by using DNA sequencing. RESULTS: among 146 ET patients, the JAK2, CALR or MPL mutations were found in: 138 cases（94.5%）including 86 cases with JAK2V617F mutation（58.9%）and 2 cases（1.4%）with exon 12 of JAK2 mutations. CALR mutations were detected in 41 cases（28.1%）, among them type 1（c.1092_1143del）in 22 cases, type 2（c.1154_1155insTTGTC）in 11 cases, and type 5（c. 1091_1142del）, type 8（c.1104_1137del）, type 41(c.1107_1137del）, type 42(c.1125_1125del）in one case respectively. In addition, 4 cases were detected withother mutations of the CALR gene（c.1107_1115del, c.1111_1144 del, c.1101 A>C, c.1112_1117del）. Moreover, 9 cases harbored MPL mutations（6.2%）. Secondly, 31 patients were detected with JAK2V617F mutation（83.8%）in 37 cases of PV, and JAK2 exon 12 mutations were found in 2 cases（5.4%）. Besides, CALR mutations were detected in 2 cases（5.4%）, including 1 case of type I, the other of novel mutation of CALR. Thirdly, 19 in 25 cases of PMF were detected with JAK2V617F mutation（76%）, 2 cases with CALR mutations（8%）. 4 patients（16%）, JAK2, CALR or MPL mutations were not detected, but among them 3 cases were found harboring other genetic abnormalities. Fourthly, no mutations of JAK2, MPL and CALR genes were detected in 124 patients with secondary thrombocytosis and 73 cases with secondary polycythemia. CONCLUSION: Combined detection of JAK2, CALR and MPL gene mutations can cover the vast majority of patients with BCR/ABL1-negative myeloproliferative neoplasms. For higher frequencies of the mutations of CALR in ET patients, CALR mutation can be used as a new diagnostic marker in ET patients with JAK2 and MPL wild type.

Four complete mitochondrial genomes of living wild-type chinese giant salamander Andrias davidianus (Amphibia: Cryptobranchidae).

The Chinese giant salamander (CGS), Andrias davidianus (Amphibian, Caudata, Cryptobranchidae), is endemic to China. After overhunting in the 1990's, it is very difficult to find the CGS in the wild. Due to mating disorder, the captive breeding population is genetically confounded. The genetic backgrounds of all wild-release individuals in China are not explicit. Herein, we reported four living wild-type complete mitochondrial genomes of this species. The gene order and contents are identical to those found in typical vertebrates. Thirteen protein-coding genes (PCGs) of 7 A. davidianus (4 from this study, 3 retrieved from GenBank) and 11 other closely species retrieved from GenBank were used to reconstruct phylogenetic tree. The Maximum likelihood (ML) topology shown that the clade of CGS has two subclades with a high support (100%). This study provides partial fundamental information for further exploring the true genetic background of whole population of A. davidianus.

Non-malignant T-cells lacking multiple pan-T markers can be found in lymph nodes.

In order to observe and ascertain the properties of a sub-group of T cells in the lymph node (LN) from seven patients who did not suffer from T cell lymphoproliferative disorders (T-LPDs), the expression levels of several pan-T markers were evaluated by multiparameter flow cytometry (FC) and the clonality of these T-cells was evaluated by both FC analysis and PCR assessment. It turned out that multiple pan-T-cell markers such as CD2, CD5 and CD7 were found to be lost in these T cells. The majority of them were positive for TCRαß, only a minority of them being positive for TCRγδ. A subset of these T-cells were positive for CD4 or CD8 or dual-negative for CD4 and CD8. Oligoclonality was detected in one case by FC, while clonal TCR rearrangement was detected in three cases. Absence of multiple pan-T-cell markers could be found in benign T cells in LNs.

Transcriptome sequencing reveals potential mechanisms of diapause preparation in bivoltine silkworm Bombyx mori (Lepidoptera: Bombycidae).

In the bivoltine strain of the silkworm, Bombyx mori, embryonic diapause is induced transgenerationally as a maternal effect. Progeny diapause ability is determined by the environmental condition such as temperature and lightness that mothers experience during their own embryonic development. Diapause preparation is a crucial phase of this process; diapause-destined individuals undergo a series of preparatory events before the entry into developmental arrest. However, the molecular regulatory mechanisms of diapause preparation have largely remained unknown. In the present study, we sequenced the transcriptome of bivoltine silkworm Qiufeng's ovaries resulted in laying of diapause destined or non-diapause eggs, using high-throughput RNA-Seq technology. Differential expression analyses identified 183 genes with higher expression, and 106 with lower expression under diapause-inducing conditions. GO and KEGG analysis revealed that the enrichment of several functional terms related to peroxisome, glycerolipid metabolism, steroid biosynthesis, longevity regulating pathway - multiple species, three signaling transductions, insect hormone biosynthesis, and cytoskeleton components. We conducted a detailed comparison of transcript profile data of ovaries from diapause-inducing and non-diapause conditions, the results imply up-regulation of peroxisomal metabolism, triacylglycerides accumulation, cryoprotectant production, and ecdysteroid biosynthesis in diapause-inducing group. Differential expression of genes related to actin cytoskeleton implies the occurrence of shifts in cellular structure and composition between diapause-inducing and non-diapause-inducing groups. The Hippo and FOXO signaling pathways may play an important role in preparing for entering diapause. This study provides an insight into the molecular events of insect diapause, in particular for the preparatory phase.

[Removal of Cd from Soil by Aspergillus fumigatus in a Semi-solid Culture].

A method was proposed to remove Cd from contaminated soils by a semi-solid culture containing Aspergillus fumigatus that have a strong resistance to Cd. The removal efficiencies of Cd in different simulated Cd pollution levels were studied and the changes in Cd adsorption and the enrichment in Aspergillus fumigatus were measured. The results showed that Aspergillus fumigatus could remove some Cd from the soil in the semi-solid culture system. When the concentration of Cd was 10 mg·kg-1, the total removal rate of Cd was up to 31%. Meanwhile, the dry weight of Aspergillus fumigatus and the pH of the system were studied during the culture process. The results showed that the dry weight decreased with the increase in culture time and Cd concentration, the maximum decrease rate of dry weight was 64%. The removal efficiency was the best when the pH was varied from 5.6 to 6.0. The changes in different extraction fractions for Cd showed that the main fractions of Cd removed by Aspergillus fumigatus were the acid-soluble fraction and the reducible fraction and the oxidizable fraction of Cd remained essentially unchanged before and after the culturing. The proposed method would provide valuable information for the remediation of heavy metal-contaminated soil by fungi.

[Effects on Physico-chemical Parameters of Glycoside Hydrolase Family 6 Genes During Composting of Agricultural Waste].

Polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) was used to study the changes of glycoside hydrolase family 6 (GH6 family) genes during the composting of agricultural waste. In addition, a redundancy analysis (RDA) and Monte Carlo permutation test were applied to determine the relationship between the changes to the GH6 family genes and physico-chemical parameters. According to the relative magnitudes of the influence caused by these different parameters on the changes to GH6 family genes, the eight-ranked physico-chemical parameters were pH > pile temperature > total nitrogen (TN) > total organic matter (TOM) > C/N ratio > moisture content > ambient temperature > water soluble carbon (WSC). The eight physico-chemical parameters, which explained 83.1% of the variation of GH6 family genes, confirmed that these parameters had important effects on the GH6 family genes changes. The pH (P=0.002), pile temperature (P=0.004), and TN (P=0.004) had the most significant impacts on these changes. These three significant factors explained 24.92%, 15.57%, and 15.04% of the variation of GH6 family genes, respectively. There were different dominant species which contain GH6 family genes in different stages of composting. The diversity and abundance of GH6 family genes demonstrated these fluctuation trends. The t-value biplots based on the RDA showed that these three significant factors had either positive or negative correlations with the dynamic changes of GH6 family genes and the microbial species contained within GH6 family genes can be generally be divided into five types. The microbial community changes are reflected by the No. 4 to No. 10 bands and these had a significant positive correlation with pile temperature and a significant negative correlation with pH and TN. The microbial community changes reflected by the No. 20 to No. 23 bands had a significant positive correlation with pH and TN.