Objective:To analyze the whole genome sequence and key site mutations of hepatitis B virus (HBV) in patients with different stages of disease progression, and to understand the relationship between HBV genetic characteristics and disease progression.Methods:Serum samples and basic information of hepatitis B patients with asymptomatic HBV carrier, chronic hepatitis B patients, cirrhosis patients and primary hepatocellular carcinoma patients were collected. Nested PCR was used to amplify the samples to obtain HBV whole gene sequences. Phylogenetic trees were constructed to determine the genotype of the samples, and gene mutations of the samples were analyzed combined with reference sequences of each type.Results:A total of 256 samples were successfully amplified, including 68 asymptomatic HBV carrier patients, 118 CHB patients, 15 LC patients and 55 HCC patients, and five genotypes (B, C, D, I and C/D) were detected. The result of comparative analysis showed that the mutation rate of 56 nucleotide sites was significantly different among the four groups ( P<0.05). In addition to the discovery of C105T, A1762T/G1764A and G1899A and other previously reported key site mutations, the mutation rates of T53A, C1485T and C1628T in newly diagnosed HCC group were significantly higher than those in other groups, and the mutation rates of T2150G and T2151C in asymptomatic HBV infection group were significantly higher than those in other groups. A total of 26 sequences were deleted, mainly distributed in the pre-C and pre-S regions. The deletion mutation rate in the HCC group was significantly higher than that in the other groups. Conclusions:The data of this study indicate that some nucleotide substitution mutations and deletion mutations may be closely related to the occurrence and development of HBV-related diseases, and HCC patients are more likely to have gene mutations than non-HCC patients. These result provide a reference for understanding the relationship between viral mutation and the progression of HBV infection-related diseases.

Objective To investigate the correlation between muscle mass and gait parameters in patients with cerebral small vessel disease(CSVD),as well as the impact of reduced muscle mass on the occurrence of falls in CSVD patients.Methods This study was employed a cross-sectional design.Ninety-five inpatients with CSVD confirmed by the Department of Neurology of the Seventh Medical Center of Chinese People's Liberation Army General Hospital from January 1,2022 to June 1,2023 were included consecutively.The 95 patients with CSVD were divided into two groups,namely the reduced muscle mass group and the normal muscle mass group,based on the criteria of appendicular skeletal muscle mass(ASM)≤7.0 kg/m2 for males and ASM ≤5.7 kg/m2 for females as reduced muscle mass.Baseline data(sex,age,years of schooling,number of accompanying diseases[hypertension,hyperlipidemia,diabetes,angina pectoris,myocardial infarction,and migraines]),cognitive function assessment results(mini-mental status examination[MMSE],verbal fluency test[VFT],clock drawing test[CDT],and trail-making test part-B[TMT-B]),gait characteristics(basic gait parameters[gait speed,stride time,stride length,stride frequency]and reanalysis gait parameters[variation coefficient of gait speed,stride time,stride length,stride frequency,and time-phase coordination index,gait asymmetry index]),CSVD imaging findings(cerebral microbleeds,lacunar infarcts,and white matter hyperintensities),and history of falls.The differences in baseline data,cognitive function assessment results,and gait characteristics between the reduced muscle mass group and the normal muscle mass group were compared and analyzed.Linear regression was used to analyze the correlation between muscle mass and gait parameters.The 95 CSVD patients were divided into fall group and non-fall group,and the differences in baseline data,cognitive function assessment results,gait characteristics,CSVD imaging findings,and muscle mass between the two groups were compared.Binary Logistic regression analysis was used to evaluate the impact of reduced muscle mass on falls.Results(1)The majority of patients in the reduced muscle mass group were females(67.7％[21/31]).There was a statistically significant difference in the sex distribution between the reduced muscle mass group and the normal muscle mass group(x2=6.143,P=0.013).There were no statistically significant differences in the other baseline characteristics and cognitive function between the two groups(all P＞0.05).(2)Compared to the normal muscle mass group,patients in the reduced muscle mass group had slower gait speed([0.72±0.16]m/s vs.[0.94±0.15]m/s),longer stride time([1.22±0.12]s vs.[1.08±0.08]s),shorter stride length([0.84±0.19]m vs.[1.00±0.14]m),and lower step frequency([100±9]steps/min vs.[112±8]steps/min).The coefficients of variation for gait speed(11.579[8.163,15.870]％vs.7.304[5.873,9.959]％),stride time(3.876[2.778,5.769]％vs.2.480[1.874,3.001]％),stride length(7.800[5.400,10.700]％vs.5.600[4.100,7.950]％),step frequency(5.313[3.568,7.272]％vs.3.674[3.099,5.082]％),and time-phase coordination index(5.894[4.392,9.080]％vs.3.828[3.031,5.972]％)were all increased,and the differences were statistically significant(all P＜0.05).There was no statistically significant difference in gait asymmetry index between the two groups(P＞0.05).Further analysis with sex and lacunar infarction as potential confounding factors showed that there were statistically significant differences in baseline gait parameters between the normal muscle mass group and the reduced muscle mass group(all P＜0.01).In the reanalysis of gait parameters,only the differences in the coefficients of variation for gait speed and stride time were statistically significant(both P＜0.05).(3)When analyzing ASM as a continuous variable,age and CDT as potential confounders,and stratifying by sex,the results showed that in male patients,baseline gait parameters(gait speed,stride time,stride length,and step frequency with 95％CI ranging from 0.057 to 0.152,-0.105 to-0.023,0.013 to 0.097,and 1.686 to 8.854,respectively),as well as coefficients of variation for stride time(95％CI-0.016 to-0.003)and stride length(95％CI-0.026 to-0.006),were correlated with muscle mass reduction(all P＜0.05).In female patients,gait speed(95％CI0.034 to 0.166)and coefficient of variation for gait speed(95％CI-0.059 to-0.010),stride time(95％CI-0.110 to-0.011),coefficient of variation for stride time(95％CI-0.025 to-0.001),and stride length(95％CI 0.018 to 0.163)were correlated with muscle mass reduction(all P＜0.05).(4)Muscle mass reduction was an independent risk factor for falls(OR,5.044,95％CI 1.840 to 13.827,P=0.002).Conclusions The preliminary analysis of this study suggests that there is a certain correlation between muscle mass and gait parameters in patients with CSVD.Additionally,the study indicates that a decrease in muscle mass among CSVD patients may increase the risk of falls.Therefore,it is important to prioritize the management of muscle mass in CSVD patients.

Objective A comparative study was conducted on rapid high-altitude models established in SD rats under two hypoxic stress modes,namely,a high-altitude field and simulated high-altitude environment,to evaluate the reliability of the simulated high-altitude test chamber.Methods SD rats were placed in a simulated rapid high-altitude animal experimental chamber(4000 m)or rapid high-altitude field laboratory(4010 m)to establish a rapid high-altitude rat model.After 24 or 72 h of exposure,physiological and pathological indicators related to high-altitude changes were collected and measured,mainly routine blood parameters,blood biochemistry,blood gas,oxidative damage indicators(superoxide dismutase(SOD),malondialdehyde(MDA),glutathione peroxidase(GSH-Px)),and inflammation indicators(interleukin 1β(IL-1 β),interferon-γ(IFN-γ),monocyte chemotactic protein 1(MCP-1)and interleukin 6(IL-6)),and pathological tissue analysis and hypoxia sensitive gene(hypoxia inducible factor-1α(Hif-1α)and vascular endothelial growth factor A(Vegfa))testing were performed.Finally,differential analysis was conducted on the result to obtain a differential evaluation report.Results At the same altitude,both high-altitude field and simulated high-altitude exposure for 72 h caused significant lung and brain damage.Under the same exposure time,the routine blood parameter,blood biochemistry,and blood gas result for the rats were similar.There were no significant differences in the detection of inflammation indicators(IL-6,IL-1β,MCP-1,and IFN-y),oxidative damage indicators(MDA,SOD,and GSH),or hypoxia-sensitive gene expression(Hif-1α and Vegfa)in the brain.However,partial pressure of carbon dioxide(PaCO2)and base excess(BE)were significantly higher in the simulated-72 h group than the other treatment group.The lung hypoxia-sensitive genes(Hif-1α and Vegfa)in the simulated-72 h group showed no significant expression difference with control group,and the brain coefficient of the high-altitude field treatment group was significantly higher than that of the simulated high-altitude treatment group.These result indicate that there may be slight differences between models prepared in high-altitude field and simulated high-altitude environments.Conclusions The simulated high-altitude animal experimental chamber can successfully establish a rapid high-altitude animal model.The simulated altitude can be appropriately increased on the basis of 4000 m.If an altitude of 4000 meters is used,the exposure time should be greater than 24 h but slightly shorter than 72 h.The simulated high-altitude experimental module has good reliability,but it is advisable to use plateaus for on-site experiments as much as possible,if conditions permit.

Objective By screening and passaging G0 generation Wistar rats,we obtained hypoxia-sensitive and hypoxia-tolerant G1 generation rats,and then the differences in hypoxia sensitivity among these rats were preliminarily explored.Methods 200 Wistar rats(half male and half female)were selected as G0 generation and placed in a controlled oxygen concentration system.The hypoxia tolerance time,which refers to the time from placement to near death,was recorded for the G0 generation rats at an oxygen volume fraction of 3％.30 rats(half male and half female)with the shortest hypoxia tolerance time were selected for mating and passage to obtain G1 generation hypoxia-sensitive rats.Similarly,30 rats(half male and half female)with the longest hypoxia tolerance time were selected for mating and passage to obtain G1 generation hypoxia-tolerant rats.An additional 24 standard Wistar rats were randomly divided into two groups:a control group and a model group,with 12 rats in each group(half male and half female).The control group was kept in a normoxic environment,while the model group,along with the G1 generation hypoxia-sensitive rats(G1 sensitive group)and G1 generation hypoxia-tolerant rats(G1 tolerant group),were placed in a hypobaric hypoxia chamber(simulating an altitude of 5 000 m).After 12 hours,various indicators,including blood gas,complete blood count,blood biochemistry,pathological sections,and hypoxia-related genes were detected or observed to compare the differences in hypoxia sensitivity among the 4 groups.Results Compared with the G0 generation standard rats,the hypoxia tolerance time of G1 generation rats was significantly prolonged(P＜0.01).Compared with the model group,the oxygen saturation(SatO2)in G1 tolerant group was significantly higher(P＜0.05).In the G1 sensitive group,the levels of white blood cell(WBC)count,neutrophil(NEUT)count,hemoglobin(HGB)concentration,hematocrit(HCT),red blood cell distribution width(RDW),platelet(PLT),and creatinine(Cr)significantly increased(P＜0.05 or P＜0.01),while actual bicarbonate(AB)content significantly decreased(P＜0.05),and the brain and lung coefficients were significantly elevated(P＜0.05).In addition,pathological section results showed that the brain and lung tissues in the model group,G1 sensitive group,and G1 tolerant group all suffered from significant damage,with no evident differences in the gene expression levels of hypoxia-inducible factor-1 α(HIF-1α)and vascular endothelial growth factor A(VEG FA)in brain tissues amongthe three groups(P＞0.05).Conclusion Compared with standard rats,G1 generation hypoxia-sensitive/tolerant rats exhibit good signs of hypoxia sensitivity/tolerance traits,but further screening and passage are still needed to purify them.

Objective To compare the organ coefficients and expressions of hypoxia-related genes in Bama and Juema pigs.Method Real-time quantitative PCR was used to detect the changes of hypoxia gene expressions in the heart,liver,spleen,lung,and kidney of Juema and Bama miniature pigs.Results The organ coefficients of kidney and spleen of Juema pigs were significantly lower than Bama miniature pigs (P<0.05 for both).The heart and lung coefficients of Juema pigs were significantly higher than that of Bama miniature pigs (P<0.05 for both).The VEGF and HIF-1α expressions in the lung and kidney in Juema pigs were significantly higher than Bama pigs (P<0.05 or P<0.01).Only the EPO expression in in the lung of Juema pigs was significantly higher than that of the Bama miniature pigs (P<0.05).Conclusions These results indicate that the variation in organ coefficients may be resulted from evolutionary factors such as adaptiveness to environmental physical and energy conditions,pathogens,and energy metabolism demands,etc.in combination.Juema miniature pigs showing a significantly higher expression of hypoxia-related genes than that in Bama minipigs indicate that it has a strong plateau adaptability by higher gene expressions.

Objective To investigate the efficacy of combining ultrasonic with three-dimensional imaging guided microwave ablation in treating large hepatocellular carcinoma.Method The clinical data of 262 patients with large unresectable liver cancer who were admitted to the Air Force General Hospital from Jan 2011 to Jun 2014 were retrospectively analyzed.Of these patients,136 underwent transcatheter arterial chemoembolization (TACE),and the remaining 126 patients underwent combined ultrasonic with three-dimensional imaging guided microwave ablation (MWA).The AFP levels,tumor ablation rate,postoperative complication rates and survival rates between the two groups were compared.Result There were significant differences in the AFP levels in the two groups before and after surgery (P ＜0.05),but no significant differences were found between the 2 groups (P ＞ 0.05).A significant difference was also observed on tumor ablation rate.In the MWA group,11 patients (8.73％) developed complications,while 21 patients (15.44％) in the TACE group developed complications,(P ＜ 0.05).The 3-month,9-month,1-year,2-year and 3-year survival rates were 99％,95％,81％,70％ and 57％ in the MWA group,and 98％,94％,63％,36％ and 28％ in the TACE group.There were significant differences in the 1-year,2-year and 3-year survival rates,but no significant difference were observed in the 3-month and 9-month survival rates.Conclnsion Combined ultrasonic with three-dimensional imaging guided microwave ablation increased the rate of tumor ablation and prolonged the survival time of patients with large hepatocellular carcinoma.

Food processing and cooking is very practical, and requires students to master certain knowledge and operational skills of nutrition. Therefore, we introduced the experience teaching design, con-crete experiment, reflective observation, abstract generalization and application, focused on practice-appli-cation-further practice in the teaching process, readjusted and set practical objectives, contents, teaching methods, evaluation, and used questionnaire and interview method to evaluate the teaching effect, which enriched the teaching content of the course and improved the effectiveness of students'!practice learning.

Objective To compare the Bama minipig and Juema minipig models of high altitude multi-organ dysfunction syndrome.Methods Six plateau-origin Juema minipigs and plain-origin Bama minpigs in each group received intravenous infusion of 0.35 mg/kg lipopolysaccharide ( LPS) , respectively.Blood samples were taken at 0 h, 3 h, 6 h, 12 h, 24 h, 48 h and 72 h after LPS infusion.Routine blood test was performed, blood CK, AST, ALT, TBIL, CRE were assayed, and histopathological examination of the lung tissues was performed at 24 h, 48 h after LPS infusion.Results The mortality of Bama minipigs was 33.3%, higher than that of 16.7%of Juema minipigs.The trend of physiological and biochemical changes was similar, but was milder in the Juema minipigs than in Bama minipigs.The lung injuries of the Bama minipigs at 24 h and 48 h were more severe than those in the Juema minipigs.Conclusions Both Bama and Juema minipig models of high altitude multi-organ dysfunction syndrome can be successfully established.Juema minipig models can be more closely and safely established, due to its own plateau biological properties, and avoid the influence by extrinsic injurious effects of plateau environment.

Objective To investigate the difference of the expression of hypoxia reaction genes of vascular endothelial growth factor(VEGF),erythropoietin (EPO) and hypoxia-inducible factor-1 alpha (HIF-1α)) at different times of rats that were induced epilepsy by kainic acid (KA),and analyze their correlation.Methods The epileptic rat model was established by intraperitoneal injection of kanic acid.The expression of VEGF,EPO and HIF-1α gene were detected by real-time fluorescent quantitative PCR assay with TaqMan probe in different times after intraperitoneal injection of KA.Results Compared with normal sodium (NS) group,the expression VEGF was higher at 12 h((8.38±1.27) ×10-3 ng/μl,P＜0.05)) and 24 h((8.30±5.08) ×10-3 ng/μl,P＜0.05)),EPO was higher at 12 h((8.42±0.90) × 10-5 ng/μl,P＜0.05)) and 48 h ((1.50±3.25) × 10-2 ng/μl,P＜0.01)) while the HIF-1α was higher at 24 h((2.11±0.21) × 10-2 ng/μl,P＜0.01)),48 h((1.50±0.33) × 10-2 ng/μl,P＜0.05))and 72 h((1.64±0.16) × 10-2 ng/μl,P＜0.01)).Furthermore,the expression of EPO showed significant correlations with HIF-1α and VEGF (r=0.573,0.471,P＜0.05),VEGF and HIF-1α had eminent correlations (r=0.803,P＜0.01).Conclusions The expression of VEGF,EPO and HIF-1α participate in the seizure procedure and there is certain correlation between the three genes.

Objective To observe the damage of liver cells and to investigate the distribution and expression of glu-cose-regulated protein 78 (Glucose regulated protein78, GRP78/Bip) in liver tissue under the positive acceleration (+Gz) exposure.Methods Totally 24 wistar rats were randomly assigned to four groups:blank control,+6Gz,+9Gz and +12Gz.Each rat was clamped to the centrifuge arm , prone position, with the head of the rat facing the axis of the centrifuge for +Gz orientation.The onset rate was +0.5 Gz/s, which was used trapezoidal acceleration curve effect and controlled by computer .Blank control group rats were placed on the arm of centrifuge and under-went a process similar to that described above , but they were not exposed to acceleration .+6Gz group,+9Gz group and +12Gz group were subjected at peak time 3 min in animal centrifuge , acceleration rate 0.5 G/s, five times with interval 30 min between times.In addition, liver tissue of rats were respectively observed by H .E.staining.Mean while, plasma aspartate aminotransferase ( AST) and alanine aminotransferase ( ALT) were tested determine the damageofliverfunction.Results +GzaccelerationstressinjuryincreasedserumASTandALTlevel.Compared with the stress control, +9Gz group and +12Gz group significantly increased in plasma ALT and AST as compared with control group ( P<0.05 ) .+12 Gz stress induced the highest level in these groups .The level of ALT in+2 Gz group was higher than that in +6 Gz group ( P<0.05 ) .HE staining showed derangement of liver cells , irregular shape, the cell gap is not clear, vacuolar changes in +Gz groups, and with the increase of G value.Compared with the control group, the expression of GRP78/Bip was focused in the cytoplasm;the expression of GRP78 in the experimental group is higher than that in the control group ( P<0.05 ) .+12 Gz group was significantly higher than+6Gz group and the control group (P<0.05).The expression of GRP78/Bip in liver tissue increased with the in-creasing of G value levels;the expression level of GRP78/Bip in +12Gz and +9Gz groups were higher than that in +6Gz and control group (P<0.05).Conclusions There is positively related expression of GRP78/Bip, which was associated with exposure of increasing G values .