Objective To investigate the inactivating effect of heat and ultraviolet(UV) light on HCV JFH-1 strain using the cell culture system. Methods The HCV JFH-1 virus stock, with an initial titer of 2.5 × 104 FFU/ml, was exposed in 56℃ water bath or to UV light for varying durations of time for explo-ring their inactivating effects on the virus. The kinetics of virus titer reduction was determined by an indirect immuno-fluorescence assay (IFA). If the cells infected with the exposed virus stock were IFA negative after three blind passages, the virus stock was considered to be inactivated completely. Results After incubation of the HCV JFH-1 virus stock (2.5 × 104 FFU/ml)in 56℃ water bath for 10 min, 20 min and 30 min, the virus titers were reduced to 1.6 × 103 FFU/ml, 3.1 × 102 FFU/ml and 3.3 × 10 FFU/ml, respectively. The exposure of the virus stock to UV light (wavelength 253.7 nm, intensity ≥60 μW/cm2, 30 cm below the UV lamp) for 15 s, 30 s and 45 s resulted in virus fiter reduction to 1.0 × 103 FFU/ml, 1.1 × 102 FFU/ml and 2.7 × 10 FFU/ml, respectively. After 40 min incubation of the virus stock at 56℃, or 1 min exposure to UV light (wavelength 253.7 nm, intensity ≥60 μW/cm2) the virus infectious titer was reduced below the detection limit of IFA, and the IFA was still negative even after three blind passages, indicating that the virus was inactivated completely. Conclusion HCV is sensitive to heat and UV light treatment. For HCV JFH-1 virus stock containing 2.5 × 104 FFU/ml virus, heat treatment at 56℃ for 40 min, or UV light expo-sure at an intensity of ≥60 μW/cm2 for 1 min, resulting in complete virus inactivation.

Objective To evaluate the efficacy and safety in the treatment of acetabular fractures via anterior approaches,including ilioinguinal,stoppa,pararectus approach.Methods Systematic literature retrieval was carried out to obtain two-arm and one-arm researches on the treatment of acetabular fractures via any one of the three approaches before May 2017,from pubmed,embase,cochrane library Databases.Data extraction and quality evaluation of studies were performed by 3 investigators independently.A meta-analysis was performed by RevMan 5.3 and Meta-Analyst beta 3.13.Results Totally 22 low to moderate quality studies,including 7 two-arm and 15 one-arm were included.There were 980 patients,including 581 in ilioinguinal approach group,264 in stoppa approach group and 135 in pararectus approach group.Meta-analysis showed the rate of anatomic reduction in stoppa approach was higher than that in ilioinguinal approach (OR=0.58,95％CI:0.36-0.94,P=0.03),which in pararectus approach was higher than that in ilioinguinal approach (0R=2.95,95％CI:1.22-7.10,P=0.02).Compared to the ilioinguinal approach,the operation time in the Stoppa approach was shorter (MD=48.01,95％CI:17.08-78.95,P=0.002),there was no statistically significant difference between pararectus approach and ilioinguinal approach.In addition,there were no statistically significant differences among three approaches in intra-operative blood loss,postoperative functional outcomes and complications.Conclusion The available limited evidence suggests that the rate of anatomical reduction in stoppa and pararectus approach may be higher than ilioinguinal approach.Compared to the ilioinguinal approach,the operation time in the stoppa approach may be shorter.In this field,further rigorous design,baseline parallel,direct comparison controlled studies on this topic are still needed.

Reversed phase high performance liquid chromatography with diode array detector was employed for simultaneous determination of five main index components and specific chromatograms analysis in Xiaochaihu granules with a linear gradient elution of acetonitrile-water (containing 0.1% phosphoric acid) as mobile phase. The results showed that five main index components (baicalin, baicalein, wogonoside, wogonin, enoxolone) were separated well under the analytical condition. The linear ranges of five components were 0.518 - 16.576, 0.069 - 2.197, 0.167 - 5.333, 0.009 - 0.297 and 0.006 - 0.270 mg x g(-1), respectively. The correlation coefficients were 0.999 9, and the average recoveries ranged from 95% to 105%. Twelve common peaks were selected as the specific chromatograms of Xiaochaihu granules with baicalin as the reference peak. There were good similarities between the reference and the ten batches of samples. The similarity coefficients were no less than 0.9. The analytical method established is highly sensitive with strong specificity and it can be used efficiently in the quality control of Xiaochaihu granules.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Flavanones ; analysis ; Flavonoids ; analysis ; Glucosides ; analysis ; Glycyrrhetinic Acid ; analysis ; Quality Control ; Sensitivity and Specificity

BACKGROUNDThe CKLF-like MARVEL transmembrane domain-containing family (CMTM) is a novel family of proteins linking chemokines and TM4SF. Different members exhibit diverse biological functions. In this study, the effect of intracellular CMTM2 on regulating human immunodeficiency virus type-1 (HIV-1) transcription was evaluated.

METHODSThe effects of CMTM2 on regulating full-length HIV-1 provirus and the HIV-1 long terminal repeat (LTR)-directed transcription were assessed by luciferase assay. Transcription factor assays, using the luciferase reporter plasmids of AP-1, CRE, and NF-κB were conducted to explore the signaling pathway(s) that may be regulated by CMTM2. The potential relationship between CMTM2 and the transcription factor AP-1 was further analyzed by Western blotting analyses to investigate the effect of CMTM2 on PMA-induced ERK1/2 phosphorylation.

RESULTSThe results from the current study revealed that CMTM2 acts as a negative regulator of HIV-1 transcription. CMTM2 exerted a suppressive action on both full-length HIV-1 provirus and HIV-1 LTR-directed transcription. Transcription factor assays showed that CMTM2 selectively inhibited basal AP-1 and CREB activity. Co-expression of HIV-1 Tat, a potent AP-1 and CREB activator, can not reverse CMTM2-mediated AP-1 and CREB inhibition, suggesting a potent and specific effect of CMTM2 on negatively regulating these two signaling pathways.

CONCLUSIONIntracellular CMTM2 can negatively regulate HIV-1 transcription, at least in part, by targeting the AP-1 and CREB pathways. Exploring the mechanisms further may lead to new ways to control HIV-1 replication.

Chemokines ; physiology ; Cyclic AMP Response Element-Binding Protein ; antagonists & inhibitors ; HIV Long Terminal Repeat ; HIV-1 ; genetics ; Humans ; Intracellular Space ; metabolism ; Jurkat Cells ; MARVEL Domain-Containing Proteins ; Tetradecanoylphorbol Acetate ; pharmacology ; Transcription Factor AP-1 ; antagonists & inhibitors ; Transcription, Genetic ; U937 Cells

Objective To assess the effect of preoperative administration of tranexamic acid (TXA) on the hidden blood loss in the surgery of intertrochanteric fractures with proximal femoral nail anti-rotation (PFNA).Methods Eighty patients with intertrochanteric fracture were treated with PFNA in our hospital from November 2015 to July 2016.They were 15 men and 65 women,with a mean age of 72.6 years.Of them,39 were included into TXA group where TXA was administered preoperatively and 41 were assigned into the control group where no TXA was used preoperatively.Blood routine examinations were carried out on one day before operation,the first and third days after operation.The surgical blood loss,operative blood transfusion,24-hour drainage after operation,and postoperative 3-day blood transfusion were recorded.The total and hidden blood losses were calculated according to the formula.The 2 groups were compared in terms of blood loss and complications.Results In TXA group,the total blood loss (1,632.3 ± 849.2 mL),the hidden blood loss (1,270.9 ± 623.3 mL) and the transfusion rate (28.2％) were significantly lower than those in the control group (2,014.8 ± 924.7 mL,1,549.1 ± 624.9 mL and 56.1％) (P ＜ 0.05).There was no significant difference between the 2 groups in visible blood loss (361.4 ± 154.3 mL for TXA group versus 465.7 ± 191.3 mL for the control group) (P ＞ 0.05).Deep venous thrombosis occurred in 2 patients,limb swelling in one patient and wound dehiscence in one patient in TXA group while deep venous thrombosis occurred in one patient,limb swelling in 3 patients and wound dehiscence in none in the control group,showing no significant differences between the 2 groups (P ＞ 0.05).No infection was observed in either group.Conclusion Preoperative administration of TXA can reduce surgical hidden blood loss and transfusion rate as well but will not increase the risk of deep vein thrombosis in the surgery of intertrochanteric fractures with PFNA.

OBJECTIVESTo evaluate the correlation between signal/cutoff (S/CO) ratios of anti-HCV EIA and their true positivity for determining the predictive value of S/CO ratios.

METHODSOne hundred and fifty-nine samples of blood from donors positive for anti-HCV at the initial screening were collected from Beijing, Guangzhou, Hangzhou, Kunming and Urumchi. All the samples were retested by Ortho and 6 Chinese domestic anti-HCV EIA kits in duplicate, and detected for HCV RNA (NAT) using Chiron Procleix HIV/HCV system (transcription mediated amplification, TMA). The HCV RNA negative samples were further tested for anti-HCV by Chiron RIBA 3.0. Either NAT or RIBA positive samples were interpreted as the true positive.

RESULTSAll 7 anti-HCV EIA kits had a significant correlation between S/CO ratios and true positivity. The S/CO ratio of Ortho > or = 3.8 predicted the true positivity in 96.1% of the samples tested. The S/CO ratios of BGI-GBI, GWK, SABC, KHB, InTec, and Wantai were > or = 7.0, > or = 10.0, > or = 6.0, > or = 10.0, > or = 8.6, > or = 14.0 and predicted 96.1%, 96.1%, 97.3%, 96.0%, 96.1%, 96.0% of the true positivity, respectively.

CONCLUSIONSThe S/CO ratios of anti-HCV EIA kits are associated with the true positivity. S/CO ratios of Ortho, BGI-GBI, GWK, SABC, KHB, InTec and Wantai predicting > or = 95% true positivity are > or = 3.8, > or = 7.0, > or = 10.0, > or = 6.0, > or = 1 0.0, > or = 8.6 and > or = 14.0, respectively.

Blood Donors ; Hepacivirus ; isolation & purification ; Humans ; Immunoenzyme Techniques ; methods ; Predictive Value of Tests ; Reagent Kits, Diagnostic ; Sensitivity and Specificity

Objective Methylglyoxal can cause the injury of human proximal tubular epithelial cell line(HK-2 cells),but the exact mechanism is still unclear. The present study aimed to explore the influence of oxidative stress and the expression of NLRP3 inflammasome in HK-2 cells induced by methylglyoxal. Methods HK-2 cells at logarithmic phase were divided into six groups:control group and 100,200,400,800,1600 μmol/L methylglyoxal groups (cells were cultured in 100,200,400,800,1600 μmol/L methylg-lyoxal concentration for 24 h). Superoxide dismutase(SOD)levels were assayed by thibabituric acid method. Release of lactate dehydro-genase(LDH)activity was detected by assay kit.ROS production was measured by DCFH-DA staining. The expression levels of NLRP3,caspase-1,IL-1β and NF-κB were evaluated by western blot. Results Compared with control group,different methylglyoxal concen-trations could enhance ROS level and LDH activity in HK-2 cells(P<0.05)and reduce SOD level significantly(P<0.05). The results of western blot showed the protein levels of NLRP3,caspase-1,IL-1β and NF-κB were significant up-regulated after the addition of methylglyoxal(P<0.05). Conclusion Methylglyoxal may induce the injury of HK-2 cells by oxidant stress and activating of NLRP3 inflammasome signaling.

Objective To compare the perioperative incidence of deep venous thrombosis (DVT) and level of plasma D-dimer between different ages in patients with lower limb fracture in Northwest China.Methods The data were retrospectively analyzed of the 1,185 patients with lower limb fracture in 5 provinces in Northwest China from July 2014 to March 2017.They were 583 males and 602 femals,with an average of 60.3 years(from 18 to 102 years).Of them,723 were from Shaanxi Province,148 from Gansu Province,137 from Qinghai Province,128 from Ningxia Hui Nationality Autonomous Regions and 49 from Xinjiang Uygur Autonomous Region.They were divided into a young group (≤44 years old) (n =288),a middle-aged group (from 45 to 59 years old) (n =256),a junior elderly group (from 60 to 74 years old) (n =263),and a senior elderly group (≥75 years old) (n =378).The incidences of perioperative deep venous thrombosis (DVT) and levels of plasma D-dimer were compared between the 4 age groups.Results The DVT incidences before and after operation in the young group were significantly lower than in the other 3 groups (P ＜ 0.05);the DVT incidence after operation in the middle-aged group was significantly lower than that in the junior elderly group (P ＜ 0.05).The pre-and post-operative levels of plasma D-dimer increased with the age of the patients;the pre-and post-operative levels of plasma D-dimer in the senior elderly group were significantly higher than in the other 3 groups (P ＜ 0.05).Conclusions The perioperative incidence of DVT was high in the elderly patients with lower limb fracture in Northwest China,peaking in the age range from 60 to 74 years old.The pre-and post-operative levels of plasma D-dimer increased with the age of the patients.

Mitochondrion is a semi-autonomous organelle, important for cell energy metabolism, apoptosis, the production of reactive oxygen species (ROS), and Ca homeostasis. Mitochondrial DNA (mtDNA) mutation is one of the primary factors in mitochondrial disorders. Though much progress has been made, there remain many difficulties in constructing cell models for mitochondrial diseases. This seriously restricts studies related to targeted drug discovery and the mechanism and therapy for such diseases. Here we summarize the characteristics of patient-specific immortalized lymphoblastoid cells, fibroblastoid cells, cytoplasmic hybrid (cybrid) cell lines, and induced pluripotent stem cells (iPSCs)-derived differentiation cells in the study of mitochondrial disorders, as well as offering discussion of roles and advances of these cell models, particularly in the screening of drugs.

Objective:To observe the effect of acupuncture on constipation in children with autism spectrum disorder (ASD). Methods:From January, 2019 to November, 2020, 50 ASD children with constipation from Rehabilitation Center of the Second Affiliated Hospital of Heilongjiang University of Chinese Medicine were admitted and randomly divided into acupuncture-rehabilitation group (n = 25) and medicine-rehabilitation group (n = 25). For constipation, the acupuncture-rehabilitation group used scalp acupuncture, body acupuncture combined with rehabilitation; the medicine-rehabilitation group took probiotics orally combined with rehabilitation. Both groups received conventional rehabilitation for constipation and the core symptoms. Constipation was treated for eight weeks, and ASD symptoms was treated for twelve weeks. After eight weeks and twelve weeks, the efficacy of constipation and the scores of main symptoms of constipation were compared, and they were also evaluated with Autism Child Behavior Scale (ABC) and Autism Treatment Evaluation Scale (ATEC). Results:Before treatment, there was no significant difference in the main symptoms of constipation, nor the scores of ABC and ATEC between two groups (P > 0.05). Eight weeks after treatment, the main symptoms of constipation significantly improved (t > 5.473, P < 0.001); twelve weeks after treatment, the efficacy of constipation and the main symptoms of constipation were better in the acupuncture-rehabilitation group than in the medicine-rehabilitation group (Z = 2.848, t = -2.139, P < 0.05). Eight weeks after treatment, the scores of ABC and ATEC significantly decreased (t > 7.139, P < 0.001), and there was no significant difference between two groups (P > 0.05); the score of behavior of ATEC significantly decreased in the acupuncture-rehabilitation group (t = 3.849, P < 0.01), and was lower than that of the medicine-rehabilitation group (t = -2.643, P < 0.05), and no significant difference in other items was found between two groups (P > 0.05); twelve weeks after treatment, the scores of ABC and ATEC were lower in the acupuncture-rehabilitation group than in the medicine-rehabilitation group (|t| > 2.156, P < 0.05). Conclusion:Acupuncture-rehabilitation can improve the longer-lasting effect of constipation, and also improve the core symptoms of ASD, which is superior to medicine-rehabilitation.