Thrombosis density ratio can predict the occurrence of pulmonary embolism and post-thrombotic syndrome in lower-extremity deep vein thrombosis patients.

OBJECTIVE: Deep vein thrombosis (DVT) formation of lower extremities can lead to serious complications including pulmonary embolism (PE) and chronic post-thrombotic syndrome (PTS). We aimed to explore the relationship between the ratio of thrombotic density and the occurrence of PE and PTS in patients with DVT of the lower extremities. METHODS: A retrospective analysis was conducted in patients who performed computed tomography venography, dividing into DVT with PE group (54 patients) and DVT-alone group (34 patients), The clinical data were recorded. Univariate and multivariate logistic regression analysis were used to analysis variables associated with PE. The ability of thrombosis density ratio and Wells score to diagnose PE was evaluated by using the receiver operating characteristic curve (ROC) area under the curve (AUC). According to the treatment and follow-up results, subgroup analysis was performed, and the Villata score was used to determine the presence or absence of PTS and its severity. RESULTS: Compare with the DVT-alone group, more patients had dyspnea and chest pain in the DVT with PE group. DVT with PE group had lower the percentage of neutrophils, white blood cell count and platelet count, while had higher blood cell count, D-dimer, wells score, thrombus and thrombus density ratio. Multivariate logistic analysis showed that percentage of neutrophils (OR(95% CIs)=1.15 (1.01,1.31), P = 0.040), platelets (OR(95% CIs)=0.96 (0.93,0.99), P = 0.011), and thrombus density ratio (OR(95% CIs)=5.99 (1.96,18.35), P = 0.002) are independent predictors of PE. The Wells score and thrombosis density ratio were consistent in the diagnostic efficacy of PE. In the subgroup analysis, there was a relevance between the ratio of thrombosis density and the Villalta score. CONCLUSION: Percentage of neutrophils, platelets, and thrombus density ratio are independent predictors of PE. The thrombosis density of DVT patients may be an index to predict the risk of PE and PTS in DVT patients.

A new species of the genus Achalinus (Squamata: Xenodermidae) from Nanning, Guangxi, China.

We describe a new species of the genus Achalinus Peters, 1869 from Daming Mountain, Shanglin County, Nanning City, Guangxi Zhuang Autonomous Region, China, based on a single adult male specimen. It can be distinguished from all the other species in Achalinus by a combination of the following morphological characters: (1) a bright yellow collar around the neck, extending forward to the ventral of the head; (2) tail length comparatively long, TaL/Tol ratio 0.25; (3) DSR 23-23-23, moderately keeled; (4) VS 3+162; (5) SC 74, unpaired; (6) cloacal plate entire; (7) SPL 6, the fourth and fifth in contact with the eye; (8) IFL 6, the first three touching the first pair of chin shields; (9) a single loreal; (10) length of suture between internasal significantly longer than that between prefrontal, LSBI/LSBP ratio 1.34; (11) two pairs of chin shields; (12) longitudinal vertebral line absent. In addition, the uncorrected p-distances between the new species and other known congeners ranged from 6.3% to 25.4% for the cytochrome c oxidase subunit 1 (CO1). With the addition of the new species the total number of described Achalinus species is increased to 23 of which 17 are found in China.

A new species of the Genus Hebius Thompson, 1913 (Squamata: Colubridae) from Baise, Guangxi, China.

A new species of the genus Hebius Thompson, 1913 is described from Youjiang District, Baise City, Guangxi Zhuang Autonomous Region, China, based on a single adult female specimen. It can be distinguished from its congeners by the following combination of characters: (1) dorsal scale rows 19-17-17, feebly keeled except the outermost row; (2) tail length comparatively long, TAL/TL ratio 0.30 in females; (3) ventrals 160 (+ 3 preventrals); (4) subcaudals 112; (5) supralabials 9, the fourth to sixth in contact with the eye; (6) infralabials 10, the first 5 touching the first pair of chin shields; (7) preocular 1; (8) postoculars 2; (9) temporals 4, arranged in three rows (1+1+2); (10) maxillary teeth 30, the last 3 enlarged, without diastem; (11) postocular streak presence; (12) background color of dorsal brownish black, a conspicuous, uniform, continuous beige stripe extending from behind the eye to the end of the tail; (13) anterior venter creamish-yellow, gradually fades to the rear, with irregular black blotches in the middle and outer quarter of ventrals, the posterior part almost completely black. The discovery of the new species increases the number of species in the genus Hebius to 51.

Upregulation of Nav1.6 expression in the rostral ventrolateral medulla of stress-induced hypertensive rats.

The rostral ventrolateral medulla (RVLM) plays a key role in mediating the development of stress-induced hypertension (SIH) by excitation and/or inhibition of sympathetic preganglionic neurons. The voltage-gated sodium channel Nav1.6 has been found to contribute to neuronal hyperexcitability. To examine the expression of Nav1.6 in the RVLM during SIH, a rat model was established by administering electric foot-shocks and noises. We found that Nav1.6 protein expression in the RVLM of SIH rats was higher than that of control rats, peaking at the tenth day of stress. Furthermore, we observed changes in blood pressure correlating with days of stress, with systolic blood pressure (SBP) found to reach a similarly timed peak at the tenth day of stress. Percentages of cells exhibiting colocalization of Nav1.6 with NeuN, a molecular marker of neurons, indicated a strong correlation between upregulation of Nav1.6 expression in NeuN-positive cells and SBP. The level of RSNA was significantly increased after 10 days of stress induction than control group. Compared with the SIHR, knockdown of Nav1.6 in RVLM of the SIHR decreased the level of SBP, heart rate (HR) and renal sympathetic nerve activity (RSNA). These results suggest that upregulated Nav1.6 expression within neurons in the RVLM of SIH rats may contribute to overactivation of the sympathetic system in response to SIH development.

[Application of damage control orthopedics for the treatment of severe multiple fractures].

OBJECTIVE: To investigate the application effect of damage control orthopedics for the treatment of severe multiple fractures. METHODS: From January 2014 to December 2016, 23 patients with severe multiple fractures were treated with the damage control orthopedics (DCO), included 14 males and 9 females with an average age of (41.57±8.29) years old ranging from 28 to 60 years old; the NISS averaged(27.70±5.44) points ranging from 18 to 40 points. As the control group, 27 patients with severe multiple fractures were treated by the early total care(ETC) technology from Jan. 2007 to Dec. 2019, included 16 males and 11 females with an average age of (38.33±9.99) years old ranging from 19 to 55 years old, the NISS averaged (31.07±6.46) points ranging from 20 to 43 points. The ICU recovery time, blood transfusion, total operation time, mortality, complication and length of hospital stay were observed and compared between two groups. RESULTS: In the DCO group, there were 22 cases surviving and 1 case death, 3 cases of postoperative complication contained 2 cases of adult respiratory distress syndrome, 1 case pin of infection in external fixation. In ETC group, there were 25 cases surviving and 2 cases death, 10 cases of postoperative complication contained 4 cases of adult respiratory distress syndrome and 3 cases of pin infection in external fixation, 1 case of wound infection and 2 cases of multiple organ failure. There was statistically significant difference between two groups in blood transfusion in operation, the ICU recovery time, and complications(P<0.05). There was no statistically significant difference in total operation time, length of hospital stay and mortality between two groups(P>0.05). CONCLUSIONS: For patients with severe multiple fractures, application of damage control orthopedics can significantly reduce the postoperative complications, ICU recovery time and intraoperative blood transfusion, provide a certain basis for clinical treatment of such patients.

[Effect of wortmannin on endothelial cell proliferation and migration induced by high glucose Müller cell conditioned medium].

OBJECTIVE: To investigate the effect of wortmannin on endothelial cells proliferation and migration induced by high glucose Müller cell conditioned medium (HGMCM). METHODS: Immunofluorescence, flow cytometry and Boyden chamber migration models were used to analyze the effect of wortmannin on endothelial cells. RESULTS: 50 nmol/L wortmannin could significantly inhibit the proliferation and migration of endothelial cells induced by HGMCM. The percentage of endothelial cells in S phase was obviously decreased [from (37.82 +/- 0.57)% to (21.91 +/- 0.23)%, P < 0.01], while there was an increase in the percentage of cells in G(0)/G(1) phase [from (54.57 +/- 1.19)% to (65.59 +/- 0.41)%, < 0.01] and G(2)/M phase (< 0.05). CONCLUSION: Wortmannin can inhibit proliferation and migration of endothelial cells induced by HGMCM, suggesting that wortmannin carries an anti-angiogenetic ability in diseased retina.

[Changes of expression of HIF-1alpha in the human retinal pigment epithelium induced by hypoxia].

OBJECTIVE: To investigate the expression of HIF-1alpha in human retinal pigment epithelium (hRPE) induced by hypoxia at different time points and to study the mechanism of choroidal neovascularization. METHODS: hRPE were isolated, cultured and identified. The changes of level of mRNA and protein of HIF-1alpha in hRPE at different time points after exposed to hypoxia were measured by semi-quantitative RT-PCR and immunofluorescence techniques. RESULTS: A significant increase of HIF-1alpha level in cultured hRPE was induced by hypoxia. After 8 hours exposed to hypoxia, the level of HIF-1alpha in the hRPE reached a peak and sustained for more than 16 hours. After a prolonged exposure to hypoxia, the morphology of hRPE began to change and the HIF-1alpha level was decreased. CONCLUSIONS: The level of HIF-1alpha in hRPE rises obviously after exposed to hypoxia. This may play a role in the occurrence of choroidal neovascularization.

Inhibition of vascular endothelial growth factor gene expression by T7-siRNAs in cultured human retinal pigment epithelial cells.

BACKGROUND: Retinal pigment epithelial (RPE) cells play an important role in the occurrence of choroidal neovascularization (CNV). Vascular endothelial growth factor (VEGF) as a positive regulatory growth factor is produced by the RPE in an autocrine or paracrine manner, promoting CNV development. Duplexes of 21 nt RNAs, known as short interfering RNAs (siRNAs), efficiently inhibit gene expression by RNA interference when introduced into mammalian cells. We searched for an efficient siRNA to interfere with VEGF expression in RPE cells and shed light on the treatment of CNV. METHODS: Human primary RPE (hRPE) cells were cultured and identified. Three pairs of siRNAs were designed according to the sequence of VEGF 1-5 extrons and synthesized by T7 RNA polymerase transcription in vitro. To evaluate the inhibitory activity of T7-siRNAs, hRPE cells were transfected via siPORT Amine. The interfering effect of T7-siRNAs in hRPE cells was examined by semiquantitative reverse transcription-polymerase chain reaction and immunofluorescence. RESULTS: Three pairs of T7-siRNAs synthesized by in vitro transcription with T7 RNA polymerase suppressed VEGF gene expression with efficiency from 65% to 90%. T7-siRNA (B), targeted region at 207 nt to 228 nt and double stranded for 21 nt with 2 nt UU 3' overhangs, was the most effective sequence tested for inhibition of VEGF expression in hRPE cells. Compared with nontransfected cells, the mean fluorescence in hRPE cells transfected with T7-sRNAs was significantly less (P < 0.01). siRNA with a single-base mismatch and ssRNA(+) did not show suppressing effect. Furthermore, it was found that siRNAs had a dose dependent inhibitory effect (5 to 10 pmol). CONCLUSION: T7-siRNA can effectively and specifically suppress VEGF expression in hRPE cells and may be a new way to treat CNV.

[Effects of high concentrations insulin on VEGF expression in cultured Müller cells].

OBJECTIVE: To investigate the effects of high concentrations of insulin on the expression of vascular endothelial growth factor (VEGF) in cultured rabbit retinal Müller cells. METHODS: Immunocytochemistry, in situ hybridization and the ELISA method were used to study the effects of different concentrations of insulin on the expression of vascular endothelial growth factor (VEGF) in cultured rabbit retinal Müller cells qualitatively and quantitatively. RESULTS: VEGF expression was enhanced obviously by the insulin, which regulated the expression of VEGF at the transcription level. CONCLUSIONS: VEGF expression in cultured Müller cells can be enhanced by high concentrations of insulin. Müller cells play a potential role in the development of neovascularization of diabetic retinopathy.

Diabetic retinopathy: VEGF, bFGF and retinal vascular pathology.

BACKGROUND: Previous research indicated that the development of diabetic retinopathy (DR) is closely related to the excessive expression of growth factors. This paper was to study the relationship of DR with vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and the retinal vascular pathological change. METHODS: Fifty-five Wistar rats, weighing 100 - 200 g, were selected and randomly divided into four groups: control group (no streptozocin injection, n = 10), M1 group (streptozocin induced diabetes for 1 month, n = 15), M3 group (streptozocin induced diabetes for 3 months, n = 15), and M5 group (streptozocin induced diabetes for 5 months, n = 15). In situ hybridization and immunohistochemistry were used to investigate the expressions of bFGF and VEGF on retinal vascular, and retinal vessels were observed by transmission electron microscope. RESULTS: There was no difference in the number of pericytes between M1 and control group (P > 0.05), but the number of pericytes decreased obviously in M3 and M5 groups compared with the control group (P < 0.01, P < 0.001, respectively). Capillary embolization and non-cell capillary were seen in M5 group. Positive expression of VEGF was found in M5 group using in situ hybridization and immunohistochemistry respectively. Positive expression of bFGF could be seen in M3 (78%) and M5 group (89%). Most remarkable changes of vessels were observed in M5 group including fragmental thickness, split of basement membrane, swelling and distortion of endothelial cells. CONCLUSIONS: In retinal vascular of the streptozocin (STZ) rats, there shows the expression of bFGF at the third month and that of VEGF at the fifth month.

[Effects of vascular endothelial growth factor and basic fibroblast growth factor in early stage diabetic retinopathy and their molecular pathological mechanism].

OBJECTIVE: To study the effects of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in early stage diabetic retinopathy and their mechanisms, so as to guide the clinical work theoretically. METHODS: Fifty-five Wistar rats were randomly divided into four groups. Ten rats were used as controls (M group). Streptozocin was injected intraperitoneally to induce diabetes for I month (M(1) group, n = 15), 3 months (M(3) group, n = 15), and 5 months (M(5) group, n = 15). At the experimental ends of each group, the rats were over-anesthetized and their eyeballs were extracted to make digest preparation. In situ hybridization and immunohistochemistry were used to investigate the expression of bFGF and VEGF on retinal vascular. Transmission electron microscopy was used to observe the histology of the retinal vessels. RESULTS: There was no difference in the number of pericytes between M(1) group and M group (P > 0.05), but the number of pericytes was significantly lower in M(3) and M(5) groups than in the M and M(1) groups (P < 0.001). VEGF in situ hybridization showed an expression rate of 34% in the M(5) group. VEGF immunohistochemistry showed an expression rate of 56% in M(5) group. bFGF in situ hybridization showed an expression rate of 78% in M(3) group and an expression rate of 89% in M(5) group. Transmission electron microscopy showed no change in M group. However, it showed swelling of endothelial cells, finger-like process into the capillary cavity, and uneven distribution of heterochromatin in pericytes in M(1) group. Obvious fragmental thickening and splitting of basement membrane, swelling and deformation, finger like process to the capillary cavity, and concentration and margination of heterochromatin in endothelial cells and swelling and deformation of rough surfaced endoplasmic reticulum and mitochondrion in pericytes were seen in the M(3) and M(5) groups, especially the latter. CONCLUSION: During the course of diabetic retinopathy, morphologic changes of vessels occurs prior to the expression of growth factors, in which the expression of VEGF follows the expression of bFGF.