Background/Aims: Existing hepatocellular carcinoma (HCC) prediction models are derived mainly from pretreatment or early on-treatment parameters. We reassessed the dynamic changes in the performance of 17 HCC models in patients with chronic hepatitis B (CHB) during long-term antiviral therapy (AVT). Methods: Among 987 CHB patients administered long-term entecavir therapy, 660 patients had 8 years of follow-up data. Model scores were calculated using on-treatment values at 2.5, 3, 3.5, 4, 4.5, and 5 years of AVT to predict threeyear HCC occurrence. Model performance was assessed with the area under the receiver operating curve (AUROC). The original model cutoffs to distinguish different levels of HCC risk were evaluated by the log-rank test. Results: The AUROCs of the 17 HCC models varied from 0.51 to 0.78 when using on-treatment scores from years 2.5 to 5. Models with a cirrhosis variable showed numerically higher AUROCs (pooled at 0.65–0.73 for treated, untreated, or mixed treatment models) than models without (treated or mixed models: 0.61–0.68; untreated models: 0.51–0.59). Stratification into low, intermediate, and high-risk levels using the original cutoff values could no longer reflect the true HCC incidence using scores after 3.5 years of AVT for models without cirrhosis and after 4 years of AVT for models with cirrhosis. Conclusions The performance of existing HCC prediction models, especially models without the cirrhosis variable, decreased in CHB patients on long-term AVT. The optimization of existing models or the development of novel models for better HCC prediction during long-term AVT is warranted.

Objective:This study retrospectively analyzed an outbreak of dengue fever in Puyang of Henan province in 2019, in order to find the sources of infection.Methods:Dengue virus IgM/IgG and NS1 antigen were tested by colloidal gold method. E gene was amplified by PCR. MegaX was used for sequences alignment to construct evolutionary distance trees.Results:After clinical and laboratory confirmation, there were 81 cases of dengue fever, 17 of which were imported case who were local farmers and worked in Combadia and Thailand, and 64 of which were indigenous cases. The E gene alignment results showed that the pathogen of this epidemic was Vietnamese 1 and highly homologous with the Vietnamese strain. After the local outbreak, dengue virus E gene developed a nucleotide site mutation which can be steadily transmission.Conclusion:The dengue fever outbreak in Puyang was a local outbreak caused by dengue virus type 1, which was associated with imported cases. Gene sequencing showed that the imported pathogen had a relatively stable and transmissible nucleotide mutation after the local epidemic.

Objective:This study retrospectively analyzed an outbreak of dengue fever in Puyang of Henan province in 2019, in order to find the sources of infection.Methods:Dengue virus IgM/IgG and NS1 antigen were tested by colloidal gold method. E gene was amplified by PCR. MegaX was used for sequences alignment to construct evolutionary distance trees.Results:After clinical and laboratory confirmation, there were 81 cases of dengue fever, 17 of which were imported case who were local farmers and worked in Combadia and Thailand, and 64 of which were indigenous cases. The E gene alignment results showed that the pathogen of this epidemic was Vietnamese 1 and highly homologous with the Vietnamese strain. After the local outbreak, dengue virus E gene developed a nucleotide site mutation which can be steadily transmission.Conclusion:The dengue fever outbreak in Puyang was a local outbreak caused by dengue virus type 1, which was associated with imported cases. Gene sequencing showed that the imported pathogen had a relatively stable and transmissible nucleotide mutation after the local epidemic.

Objective:To investigate the causes of immune failure in the population with high vaccination rate of measles and rubella vaccine in Beijing by detecting the IgG antibody affinity in suspected cases of measles and rubella.Methods:Serum samples of 276 suspected cases of measles and rubella were tested for IgM and IgG antibodies by enzyme-linked immunosorbent assay (ELISA). The affinity of IgG antibody was detected, and the relative affinity index was calculated.Results:Among the 276 suspected cases, 104 were measles and 108 were rubella. Six measles cases had vaccination history and were caused by primary immunization failure ( n=3) and secondary immunization failure ( n=3). Twelve rubella cases had vaccination history and were due to primary immunization failure ( n=4) and secondary immunization failure ( n=8). Specific high-affinity antibodies were detected in nine measles cases and seven rubella cases without vaccination history, which indicated that these cases were reinfected. In the cases without measles or rubella, other pathogenic infections including mixed infections were detected, which were mainly caused by EB virus. Conclusions:Both primary and secondary immunization failure occurred in the population with immunization history. Reinfection was found in the patients who had not received vaccination against measles or rubella. Other pathogenic infections were existed among the cases without measles or rubella. Thus, misdiagnosis was responsible for the increased proportion of measles and rubella patients with immunization history in suspected cases in recent years. Full-course vaccination was conducive to produce high-affinity antibodies against measles and rubella. A supplementary vaccination campaign should be launched to consolidate the immune barrier against measles and rubella in key population or high-risk population, aiming to block the circulation of measles virus and achieve the goal of eliminating measles.

Objective:To investigate the value of CT angiography (CTA) point sign in the treatment of patients with moderate amount basal ganglia hemorrhage at ultra-early stage by trephination and drainage or craniotomy, and its influence in the prognoses.Methods:One hundred and twenty-six patients with moderate amount basal ganglia hemorrhage (30-60 mL) admitted to our hospital from March 2017 to March 2019 were chosen in our study; these patients were evaluated and conformed to have the same tolerance of craniotomy or drainage; and their families agreed to the ultra-early surgical treatments; their clinical data were retrospectively collected. They all accepted CTA before operation. Among them, 68 were into the craniotomy group, including 38 into CTA spot sign negative sub-group and 30 into positive sub-group; 58 were into the trephination and drainage group, including 39 into CTA spot sign negative sub-group and 19 into positive sub-group. The differences of favorable prognosis rate and postoperative re-hemorrhage rate were compared between the craniotomy group and trephination and drainage group, as well as each two sub-groups.Results:The favorable prognosis rate and postoperative re-hemorrhage rate of patients in the craniotomy group (61.8% and 2.9%) were significantly lower as compared with those in the trephination and drainage group (82.8% and 15.5%, P<0.05). In the craniotomy group, the favorable prognosis rate and postoperative re-hemorrhage rate in the CTA spot sign positive sub-group (60.0% and 4.8%) were higher than those in the negative sub-group (63.2% and 2.1%), without significant differences ( P>0.05); in the trephination and drainage group, the favorable prognosis rate and postoperative re-hemorrhage rate in the CTA spot sign positive sub-group (63.2% and 36.8%) were significantly different as compared with those in the negative sub-group (92.3% and 5.1%, P<0.05). Conclusion:Among patients with moderate amount basal ganglia hemorrhage, prognoses can be effectively improved in the following treatments: if the patients have negative CTA spot sign, are evaluated to have low risk of postoperative re-hemorrhage after craniotomy or drainage, and are considered that the prognosis by drainage is better than that by craniotomy, trephination and drainage should be selected; if the patients have positive CTA spot sign, and are evaluated to have lower risk of postoperative re-hemorrhage by craniotomy than that by drainage, craniotomy should be selected.

An in vitro blood-brain barrier (BBB) model is critical for enabling rapid screening of the BBB permeability of the drugs targeting on the central nervous system. Though many models have been developed, their reproducibility and renewability remain a challenge. Furthermore, drug transport data from many of the models do not correlate well with the data for in vivo BBB drug transport. Induced-pluripotent stem cell (iPSC) technology provides reproducible cell resources for in vitro BBB modeling. Here, we generated a human in vitro BBB model by differentiating the human iPSC (hiPSC) line GM25256 into brain endothelial-type cells. The model displayed BBB characteristics including tight junction proteins (ZO-1, claudin-5, and occludin) and endothelial markers (von Willebrand factor and Ulex), as well as high trans-endothelial electrical resistance (TEER) (1560 Ω.cm ± 230 Ω.cm) and γ-GTPase activity. Co-culture with primary rat astrocytes significantly increased the TEER of the model (2970 Ω.cm to 4185 Ω.cm). RNAseq analysis confirmed the expression of key BBB-related genes in the hiPSC-derived endothelial cells in comparison with primary human brain microvascular endothelial cells, including P-glycoprotein (Pgp) and breast cancer resistant protein (BCRP). Drug transport assays for nine CNS compounds showed that the permeability of non-Pgp/BCRP and Pgp/BCRP substrates across the model was strongly correlated with rodent in situ brain perfusion data for these compounds (R = 0.982 and R = 0.9973, respectively), demonstrating the functionality of the drug transporters in the model. Thus, this model may be used to rapidly screen CNS compounds, to predict the in vivo BBB permeability of these compounds and to study the biology of the BBB.

Objective To observe the clinical effects of arthroscopic reconstruction for anterior cruciate ligament ( ACL ) tear using tension-relieving technique and enhanced recovery after surgery ( ERAS ) . Methods Between May 2014 and June 2016, 80 patients with ACL tear were randomly di-vided into 2 equal groups. The experimental group was treated with arthroscopic reconstruction using hamstring tendon autograft and tension-relieving technique followed by ERAS; the control group was treated with arthroscopic reconstruction using hamstring tendon autograft only followed by conventional postoperative re-covery. The 2 groups were compared in terms of knee flexion angles at postoperative 2nd, 4th and 8th weeks, and the IKDC ( International Knee Documentation Committee ) , HSS ( Hospital for Special Surgery ) , and Lysholm scores at postoperative 3rd, 6th and 12th months. Results The 80 patients obtained an average follow-up of 12. 3 ± 1. 7 months. Arthroscopy one year postoperation revealed fine ACL growth without laxity or other complications in the experimental group. Joint stiffness appeared in one case at 2 months postoperation and ACL laxity was observed in 3 cases by arthroscopy at 12 months postoperation in the control group. The knee flexion angles at postoperative 2nd, 4th and 8th weeks, and the IKDC, HSS and Lysholm scores at postoperative 3rd, 6th and 12th months in the experimental group were significantly better than those in the control group ( P <0. 05 ) . In both groups, the IKDC, HSS and Lysholm scores at postoperative 3rd, 6th and 12th months and the knee flexion angles at postoperative 2nd, 4th and 8th weeks were all significantly im-proved than the preoperative values ( P <0. 05 ) . Conclusion In arthroscopic reconstruction for ACL tear, tension-relieving technique combined with ERAS can promote functional recovery of the knee and reduce postoperative complications, facilitating early recovery of sports function.

Objective To observe the effects of perfusion of rosiglitazone (RSG) in lesion areas on the expression levels of the perihematomal tight junction-associated proteins occludin and zonula occluden-1 (ZO-1) mRNA,the permeability of blood-brain-barrier (BBB),and neurological function score in a rabbit model of cerebral hemorrhage (ICH).Methods A total of 45 healthy male rabbits were selected (a body mass of 2.0 to 2.5 kg).They were divided into 3 groups,a control group,a ICH model group,and a RSG treatment group (n =15,5 of them for BBB determination) according to the random number table.The control group was use to simulate the process of making intracranial hematoma.After successful puncture,the target was iujected with isotonic saline 0.3 ml and isotonic saline 0.1 ml was injected again after 6 h;after successful puncture,the ICH model group was injected with 0.3 ml of autologous non-anticoagulant arterial blood,and the target was injected into isotonic saline 0.1 ml after 6 h;RSG 0.5 mg was infused into the hematoma area (dissolved in 0.1 ml isotonic saline) in the RSG treatment group at 6 h after the ICH model was successfully induced.All rabbits in each group were sacrificed on day 7 after the neurological deficit scale score (Purdy score).Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression levels of perihematomal oecludin and ZO-1 mRNA.The formamide method was used to measure the Evans blue (EB) content in the perihematomal tissue in order to evaluate the permeability of BBB.Results (1) Neurological function scores:Purdy scores of the control group,ICH model group,and RSG treatment group were 2.53 ± 0.05,8.13 ± 0.06),and 6.67 ± 0.08,respectively.There were significant differences among the groups (F =459.116,P ＜ 0.01).Compared with the control group,Purdy scores of the ICH model group and RSG treatment group were increased significantly (all P ＜ 0.01).Compared with the ICH model group,Purdy scores of the RSG treatment group were decreased (P ＜ 0.05).(2) The expression levels of occludin and ZO-1 mRNA:The differences were statistically significant in occludin and ZO-1 mRNA in the control group,ICH model group,and RSG treatment group (1.013 ±0.051,1.001 ± 0.045;0.221 ± 0.017,0.247 ± 0.019;0.498 ± 0.041,and 0.613 ± 0.045,respectively in each group;F =443.924 and 381.929 respectively,all P ＜ 0.01).Compared with the control group,the expression levels of occludin and ZO-1 mRNA were significantly decreased in the ICH model group and RSG treatment group (all P ＜ 0.01).Compared with the ICH model group,the expression levels of occludin and ZO-1 mRNA were increased in the RSG treatment group (all P ＜ 0.05).(3) The permeability of BBB:The EB content in the control group,ICH model group,and RSG treatment group were 12.0 ± 1.0,51.6 ± 0.9,and 36.4 ± 1.0 μg/g,respectively.The differences were statistically significant among the groups (F =223.516,P ＜ 0.01).Compared with the control group,the EB content was significantly increased in the ICH model group and RSG treatment group (all P ＜ 0.01).Compared with the model group,the EB content was significantly decreased in the RSG treatment group (P ＜ 0.01).Conclusion The perfusion of RSG in the lesion area can significantly improve the neurological function of rabbits after ICH,increase the expression levels of occludin and ZO-1 mRNA in the perihematomal tissue,and decrease the permeability of BBB.

Objective To observe the effects of hydrogen sulfide (H2S) on structure and function of mitochondria of lung in rats with acute lung injury (ALI) induced by lipopolysaccharide (LPS).Methods Forty healthy male Sprague-Dawley (SD) rats were randomly divided into control group, LPS injury group, and low-, middle-, and high-dose NaHS groups, with 8 rats in each group. The rats in LPS injury group were given LPS 5 mg/kg via sublingual vein, and those in low-, middle-, and high-dose NaHS groups were challenged by LPS for 3 hours followed by intraperitoneally injection of 0.78, 1.56 and 3.12 mg/kg NaHS respectively in a volume of 2 mL/kg. The rats in control group were given 2 mL/kg normal saline via sublingual vein. The rats were sacrificed at 6 hours after model reproduction, and the lung tissues were harvested on time. The mitochondria in lung tissues were isolated with differential centrifugation. The lung mitochondria ultra structures were observed with electron microscope. The content ofmalondialdehyde (MDA) in mitochondria was determined with thiobarbituric acid method, and the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and adenosine triphosphatase (ATPase) were determined with xanthine oxidase method. The mitochondrial activity and swelling were determined by multiskan spectrum.Results It was shown by transmission electron microscope that the mitochondrial structure in the control group was normal. The mitochondria in rat lung cells was swollen with disrupted or disintegrated cristae, the osmiophilic lamellar bodies had fused or disappeared, and rough endoplasmic reticulum degranulation phenomenon was obvious in LPS injury group. The mitochondrial damage was slightly mitigated in the low-dose NaHS group, and it was significantly mitigated in the middle-dose and high-dose NaHS groups. Compared with control group, the MDA content in lung mitochondria in LPS injury group was significantly increased (nmol/mg: 26.30±1.45 vs. 11.16±1.20), andSOD, GSH-Px, and ATPase activities were significantly decreased [SOD (U/mg): 18.78±1.13 vs. 27.44±1.97, GSH-Px (U/mg): 63.91±1.99 vs. 128.15±3.47, ATPase (U/mg): 4.83±0.25 vs. 9.92±0.65]; as well as the activity of the mitochondria was significantly decreased (A value: 0.164±0.025 vs. 0.319±0.045), and the swelling of the mitochondria was significantly increased (A value: 0.182±0.012 vs. 0.273±0.023), all with significantly statistical differences (allP < 0.01). Compared with LPS injury group, the MDA contents in low-, middle-, and high-dose NaHS groups were significantly decreased (nmol/mg: 21.89±1.23, 17.63±1.56, 12.19±1.30 vs. 26.30±1.45), and the SOD, GSH-PX, and ATPase activities were significantly increased [SOD (U/mg): 20.13±0.85, 21.38±1.22, 24.05±1.56 vs. 18.78±1.13; GSH-Px (U/mg): 82.06±1.65, 101.45±2.14, 117.80±2.12 vs. 63.91±1.99; ATPase (U/mg): 5.34±0.23, 7.06±0.37, 8.78±0.44 vs. 4.83±0.25]; as well as the activity of the mitochondria was markedly increased (A value: 0.194±0.018, 0.230±0.032, 0.297±0.038 vs. 0.164±0.025), and the swelling of mitochondria was markedly decreased (A value: 0.195±0.008, 0.219±0.017, 0.249±0.018 vs. 0.182±0.012), all with significantly statistical differences (allP < 0.05). Moreover, the protective effect of NaHS showed a dose-dependent manner.Conclusion It could be concluded that LPS induce mitochondrial structural damage and functional impairment in rats with ALI induced by LPS, and H2S have a beneficial effect against ALI induced by LPS with decreasing the mitochondrial lipid peroxidation level and protecting the cell structure and function, and the effect is correlated with the dosage.

Objective To evaluate the effect of exogenous hydrogen sulfide (H2S) on mitochondrion-dependent apoptosis in lung tissues of rats with endotoxemia.Methods Forty healthy male SpragueDawley rats,aged 2-3 months,weighing 250-310 g,were divided into 5 groups (n=8 each) using a random number table:control group (group C),lipopolysaccharide (LPS) group,low dose sodium hydrosulphide (NaHS) group (group L-NaHS),moderate dose NaHS group (group M-NaHS) and high dose NaHS group (group H-NaHS).Endotoxemia was induced by Ⅳ LPS 5 mg/kg in chloral hydrate-anesthetized rats.The equal volume of 0.9％ sodium chloride solution was intravenously injected in group C.NaHS (an exogenous donor of H2S) 0.78,1.56 and 3.12 mg/kg were intraperitoneally injected at 3 h after LPS injection in L-NaHS,M-NaHS and H-NaHS groups,respectively.The rats were sacrificed at 6 h after injection of LPS or 0.9％ sodium chloride solution,and lungs were removed for examination of the mitochondrial ultrastructure of lung tissues and for determination of apoptosis in lung cells (by flow cytometry) and expression of caspase-3,caspase-9,Bcl-2 and Bax protein and mRNA in lung tissues (by Western blot or real-timne polymerase chain reaction).The apoptosis rate and ratio of Bcl-2 expression to Bax expression (Bcl-2/Bax ratio) were calculated.The expression of cytochrome c (Cyt c) in cytoplasm and mitochondria of lung tissues was detected by Western blot.Results The apoptosis rate was significantly increased,the expression of caspase-3,caspase-9,Bcl-2 and Bax protein and mRNA was up-regulated,Bcl-2/Bax ratio was decreased,the expression of Cyt c in cytoplasm was up-regulated,and the expression of Cyt c in mitochondria was down-regulated in group LPS (P ＜0.05 or 0.01).Compared with group LPS,the apoptosis rate was significantly decreased,the expression of caspase-3 and Bax protein and mRNA was down-regulated,the expression of Bcl-2 protein and mRNA was up-regulated,Bcl-2/Bax ratio was increased,the expression of Cyt c in cytoplasm was down-regulated,and the expression of Cyt c in mitochondria was up-regulated in L-NaHS,M-NaHS and H-NaHS groups,the expression of caspase-9 protein and mRNA was significantly down-regulated in M-NaHS and H-NaHS groups,and the expression of caspase-9 was significantly down-regulated (P ＜0.05 or 0.01),and no significant change was found in caspase-9 mRNA expression in group L-NaHS (P＞0.05),and the damage to mitochondrial ultrastructure was significantly mitigated in MNaHS and H-NaHS groups.Conclusion The mechanism by which exogenous H2S inhibits cell apoptosis in lung tissues may be related to inhibition of mitochondrion-dependent apoptosis in rats with endotoxemia.