[A clinical study of high dependency units to reduce ICU readmission rates in patients with severe liver disease].

Objective: To investigate the real-world difference in the ICU readmission rate between the high-dependency unit (HDU) and the general ward so as to reflect the role of HDU in the diagnosis and management of patients with SLD. Methods: Patients with severe liver disease who were consecutively enrolled were step-downed to HDU and general wards in the ICU of the Fifth Medical Center of the People's Liberation Army General Hospital between July 2017 and December 2021. The main liver function indicators, MELD scores, and other were compared between the two groups. SLD severity, ICU readmission rates, and others differences were analyzed among the patients transferred to different wards. The HDU role was clarified for SLD patients' grade management. The area under the curve of the receiver operating characteristic curve (AUROC) was used to calculate and explore the feasibility of a baseline Model for End-Stage Liver Disease (MELD) score to define the treatment scope of HDU. Results: The SLD group of patients who were transferred to HDU had significantly higher levels of the international normalized ratio, bilirubin, alanine aminotransferase, MELD score, and other factors compared to those in the general ward (P < 0.05). 70.7% of SLD patients in the HDU group had a MELD score > 17, while 61.9% of SLD patients in the general ward group had a MELD score ≤ 17. The overall ICU readmission rate in this cohort was 11.4%. The ICU readmission rate was significantly higher with a MELD score of > 23 (20.0%) than that with a MELD score of ≤ 23 (8.6%) in patients with SLD, according to the MELD score quartile P75 (P = 0.020). The ICU readmission rate was 8.2% when MELD score ≤ 23, and 9.1% when MELD score>23 in the HDU group, with no statistically significant difference (P = 1.000). However, in the general ward group, the ICU readmission rate in patients with a MELD score ≤ 23 was 8.8%, and when the MELD score was >23, the ICU readmission rate significantly increased to 36.4% (P = 0.001). The optimal cut-off value of the MELD score for predicting ICU readmission in patients with SLD in the general ward group was 23.5. Conclusion: The high-dependency unit can better undertake ICU step-down patients with SLD and significantly reduce the ICU readmission rate with MELD scores > 23 in practice. Additionally, ICU step-down SLD patients with a MELD score > 23 are suitable for transfer to HDU treatment.

[Analysis of the clinical predictive value of lactate on the prognosis of patients with acute-on-chronic liver failure combined with infection].

Objective: To explore the predictive value of lactic acid for the adverse prognostic outcomes in patients with acute-on-chronic liver failure combined with infection. Methods: A retrospective analysis was conducted on the clinical data of 208 cases of ACLF combined with infection who were hospitalized from January 2014 to March 2016. Patients were divided into a survival group (n = 83) and a mortality group (n = 125) according to the results of a 90-day follow-up. The clinical data were statistically analyzed between the two groups. Multivariate logistic regression with two categorical variables was used to analyze the independent risk factors for 90-day disease mortality and establish a new prediction model. The receiver operating characteristic curve (ROC curve) was used to evaluate the predictive value of lactic acid, the MELD score, the MELD-Na score, lactic acid combined with the MELD score, lactic acid combined with the MELD-Na score, and the new model. Results: The 90-day mortality rate of 208 cases of ACLF combined with infection was 60.1%. There were statistically significant differences in white blood cell count, neutrophil count, total bilirubin (TBil), serum creatinine (Cr), blood urea nitrogen (BUN), blood ammonia, the international normalized ratio (INR), lactic acid (LAC), procalcitonin, the MELD score, the MELD-Na score, hepatic encephalopathy (HE), acute kidney injury (AKI), and bleeding between the two groups. Multivariate logistic regression analysis showed that TBil, INR, LAC, HE, and bleeding were independent risk factors for 90-day mortality in patients with ACLF combined with infection. After the establishment of MELD-LAC, MELD-Na-LAC, and a new prediction model, the ROC curve revealed that the AUC (95% confidence interval) of MELD-LAC and MELD-Na LAC were 0.819 (0.759 ~ 0.870) and 0.838 (0.780 ~ 0.886), respectively, and was superior than the MELD score [0.766 (0.702 ~ 0.823)] and MELD-Na score [0.788 (0.726 ~ 0.843)], with P < 0.05, while the new model had an AUC of 0.924, the sensitivity of 83.9%, specificity of 89.9%, and accuracy of 87.8%, which was higher than LAC, MELD score, MELD-Na score, MELD-LAC, and MELD-Na-LAC (P < 0.01). Conclusion: Lactic acid is an independent risk factor for mortality in patients with ACLF combined with infection, and it improves the clinical predictive value of MELD and MELD-Na for the prognosis of mortality.

[Recompensation of complications in patients with hepatitis B virus-related decompensated cirrhosis treated with entecavir antiviral therapy].

Objective: To analyze the occurrence of recompensation conditions in patients with chronic hepatitis B virus-related decompensated cirrhosis after entecavir antiviral therapy. Methods: Patients with hepatitis B virus-related decompensated cirrhosis with ascites as the initial manifestation were prospectively enrolled. Patients who received entecavir treatment for 120 weeks and were followed up every 24 weeks (including clinical endpoint events, hematological and imaging indicators, and others) were calculated for recompensation rates according to the Baveno VII criteria. Measurement data were compared using the Student t-test or Mann-Whitney U test between groups. Categorical data were compared by the χ (2) test or Fisher's exact probability method between groups. Results: 283 of the 320 enrolled cases completed the 120-week follow-up, and 92.2% (261/283) achieved a virological response (HBV DNA 20 IU/ml). Child-Pugh and MELD scores were significantly improved after treatment (8.33 ± 1.90 vs. 5.77 ± 1.37, t = 12.70, P < 0.001; 13.37 ± 4.44 vs. 10.45 ± 4.58, t = 5.963, P < 0.001). During the 120-week follow-up period, 14 cases died, two received liver transplants, 19 developed hepatocellular cancer, 11 developed gastroesophageal variceal bleeding, and four developed hepatic encephalopathy. 60.4% (171/283) (no decompensation events occurred for 12 months) and 56.2% (159/283) (no decompensation events occurred for 12 months and improved liver function) of the patients had achieved clinical recompensation within 120 weeks. Patients with baseline MELD scores > 15 after active antiviral therapy achieved higher recompensation than patients with baseline MELD scores ≤15 [50/74 (67.6%) vs. 109/209 (52.2%), χ (2) = 5.275, P = 0.029]. Conclusion: Antiviral therapy can significantly improve the prognosis of patients with hepatitis B virus-related decompensated cirrhosis. The majority of patients (56.2%) had achieved recompensation. Patients with severe disease did not have a lower probability of recompensation at baseline than other patients.

[Minimally invasive liver surgery:progress in the last decade and evolving trends].

With the promotion and popularity of minimally invasive surgery and instruments,minimally invasive technologies have been widely used in the diagnosis and treatment of liver disease. In the past decade,with the development of relevant instruments,improvement of skills,and perfection of theories,the concept of individualized minimal invasion and precision has been popularized. Minimally invasive liver surgery is moving toward the goal of innovation-driven high-quality development.

[Distribution and reference intervals of daytime intraocular pressure in the eye health screening population of Handan].

Objective: To describe the distribution and establish reference intervals (RI) of daytime intraocular pressure (IOP) in the eye health screening population of Handan. Methods: This cross-sectional study included subjects who participated in eye health screening at the Physical Examination Center of Handan First Hospital from May 2021 to June 2022. A complete general and ocular examination was performed, including measurements of visual acuity and IOP (using Goldmann tonometry), slit lamp microscopy, fundus photography, and anterior and posterior segment optical coherence tomography. Subjects with factors that could cause significant changes in IOP or affect the accuracy of IOP measurement, or with an inability to measure IOP were excluded. Simple random sampling was used to select participants, who were grouped by gender and age (18 to <30, 30 to <40, 40 to <50, 50 to <60, 60 to <70, and ≥70 years). Central corneal thickness and IOP at 8 to 11 o'clock in one eye of each participant were recorded. The independent sample t test and ANOVA were used for statistical analysis, and the RI of IOP values was calculated by x¯±1.96s. Results: A total of 9 310 subjects had their IOP measured, and 3 491 participants (3 491 eyes) were randomly selected from 7 886 healthy subjects. The age of the participants was (47.74±14.47) years old, ranging from 18 to 90 years old. There were 1 694 males and 1 797 females. The central corneal thickness of all participants was (525.56±49.39) µm. The daytime IOP of all participants was (15.40±2.54) mmHg (1 mmHg=0.133 kPa), and the RI was 10.42 to 20.39 mmHg. The IOP was (15.49±2.58) mmHg for males and (15.29±2.49) mmHg for females, and the gender difference was statistically significant (P<0.05). The RI of daytime IOP values was 10.43 to 20.54 mmHg for males and 10.41 to 20.18 mmHg for females. There were significant differences in daytime IOP [(15.13±2.58), (15.33±2.53), (15.49±2.50), (15.53±2.55), (15.39±2.62), and (15.28±2.52) mmHg] among 6 age groups (P<0.05). Conclusions: The distribution of daytime IOP in different gender and age groups in the eye health screening population of Handan and the RIs derived from the distribution were roughly the same as the international normal IOP RI (10 to 21 mmHg). It is recommended to refer to the RI of daytime IOP values of different genders and ages for clinical decision.

[Investigation on status of dyslipidemia in Chinese females aged 35 years or above].

Objective: To investigate the prevalence, awareness, treatment and control status of dyslipidemia among females aged ≥35 years old across China. Methods: Participants were selected by stratified multistage random sampling method in the "Twelfth Five-Year Plan" National Science and Technology Support Project "Survey on the Prevalence of Important Cardiovascular Diseases and Key Technology Research in China" project. This study is a retrospective, cross-sectional study. A total of 17 418 females aged 35 years and over were included in the current study. The basic information such as age, medical history and menopause was collected by questionnaire. The blood lipid parameters were derived from clinical laboratory examinations. The prevalence of dyslipidemia and the rate of awareness, treatment, and control of dyslipidemia were analyzed in females aged 35 years and over. Results: The age of participants was (56.2±13.0) years old, and the prevalence of dyslipidemia was 33.1% (5 765/17 418). The prevalence rates of high total cholesterol, hypertriglyceridemia, low HDL-C and high LDL-C were 9.7% (1 695/17 418), 11.1% (1 925/17 418), 10.9% (1 889/17 418) and 7.3% (1 262/17 418), respectively. The prevalence of dyslipidemia increased with age and the prevalence of dyslipidemia in women who were not married, Han, menarche age>16 years, obesity, central obesity, alcohol consumption, diabetes, hypertension and family history of cardiovascular disease were higher than those without such characteristics (P<0.05). There were 10 432 (59.9%) menopausal females in this cohort and prevalence of dyslipidemia of these participants was 38.8% (4 048/10 432), which was higher than that of non-postmenopausal females (24.6%, 1 717/6 986) (P<0.05). The awareness rates, treatment rates and control rates of dyslipidemia were 33.9% (1 953/5 765), 15.1% (870/5 765) and 2.5% (143/5 765) respectively among females aged 35 years and over in China. Conclusion: The prevalence of dyslipidemia in Chinese females aged 35 years and over is high, and its awareness, treatment, and control rates need to be optimized.

[Resting-state functional magnetic resonance imaging analysis of abnormal directional functional connectivity of the nucleus accumbens in patients with chronic tinnitus].

Objective: To investigate abnormal directional functional connectivity of the nucleus accumbens (NAc) in chronic tinnitus patients using resting-state functional magnetic resonance imaging (fMRI), and to determine the relationship between the degree of this connectivity and tinnitus characteristics. Methods: The resting-state fMRI data of 40 patients with bilateral chronic tinnitus (12 males and 28 females, aged from 26 to 63(50.6±11.6) years) and 40 healthy controls with normal hearing (16 males and 24 females, aged from 26 to 70(45.9±12.4)years) were retrospectively enrolled from the Department of Otolaryngology, Nanjing First Hospital from January 2017 to January 2020. The bilateral NAc were selected as seeds to detect the directional functional connectivity with the whole brain, then the effective connectivity values between the two groups were compared using Granger Causality Analysis (GCA), and the correlation between the effective connectivity and the characteristics of tinnitus was calculated. Results: Compared with healthy controls, the effective connectivity from the left NAc to left middle frontal gyrus in patients with bilateral chronic tinnitus was increased [(1.0±0.2)vs(0.6±0.3)], the effective connectivity from the right NAc to left inferior frontal gyrus was enhanced [(0.9±0.3)vs(0.6±0.4)], the effective connectivity from the right middle temporal gyrus to left NAc was enhanced [(1.0±0.2)vs(0.5±0.3)], the effective connectivity from the right middle frontal gyrus to right NAc was also enhanced[(1.0±0.2)vs(0.5±0.3)](all P<0.05). After adjusting for age, gender, education level, and gray matter volume, positive correlations was observed between the Tinnitus Handicap Questionnaire (THQ) scores and increased effective connectivity values from the left NAc to the left middle frontal gyrus (r=0.386, P=0.020). Additionally, enhanced effective connectivity values from the right middle frontal gyrus to the right NAc was also positively associated with tinnitus duration (r=0.390, P=0.019). Conclusion: The directional functional connectivity between the NAc and prefrontal cortex in patients with chronic tinnitus is enhanced.

[Navigation-guided nasal endoscopy for removal of the cavernous hemangioma of the orbital apex through the sphenoid approach].

Objective: To explore the feasibility of navigation-guided nasal endoscopy for removal of the cavernous hemangioma of the orbital apex through the sphenoid approach. Methods: Retrospective case series study. From May 2012 to December 2019, 12 patients (12 eyes) with imaging findings of cavernous hemangioma in the orbital apex were collected at the Eye Hospital Affiliated to Nanchang University, including 3 males and 9 females aged 32 to 59 years. All patients underwent navigation-guided sinusoscopy through the sphenoid approach to remove the cavernous hemangioma of the orbital apex (video attached). Changes of visual function and complications after operation were analyzed. Results: In 3 patients without visual impairment, the postoperative visual function was still normal. Among the remaining 9 patients with preoperative visual impairment, visual function was fully recovered in 3 patients after operation, was improved in 2 patients, and had no change in 4 patients. There were no complications in 3 of the 12 patients, and 9 patients had transient, mildly limited intraocular rotation with diplopia after operation, which all returned to normal within 1 month. Conclusion: Navigation-guided sinus endoscopy through the sphenoid approach is effective and feasible in the removal of the cavernous hemangioma of the orbital apex. (Chin J Ophthalmol, 2021, 57: 837-843).

[Clinical characteristics of infection-induced acute renal injury in acute-on-chronic liver failure].

Objective: To investigate the incidence rate, influencing factors and prognosis of infection-induced acute renal injury (AKI) in patients with acute-on-chronic liver failure (ACLF). Methods: 516 cases with acute-on-chronic liver failure complicated with infection that were hospitalized in our hospital during 2014 to 2016 were retrospectively studied. General conditions and clinical characteristics of the patients were collected, and grouped according to the presence or absence of incidence and severity of AKI. General conditions, laboratory results, occurrence of complications and survival were compared and analyzed. Results: The main causes were HBV infection (67.8%) and alcoholic liver disease (20.0%). The most common sites of infection were abdominal cavity, lung and blood. Multivariate analysis showed that neutrophil count, TBIL, lactate and septic shock were independent risk factors for infection-induced AKI in ACLF patients. The cumulative mortality in patients with AKI after infection at 28, 90 and 360 days was significantly higher than those without AKI (51.6% and 20.5%, 70.2% and 40.3%, 73.4% and 45.9%; P < 0.01). In both groups, deaths had occurred mainly in the early (0 ~ 28 d) and middle (29 ~ 90 d) stage of follow-up period. In the late follow-up period (91-360 d), there was no statistically significant difference in mortality rate between the two groups. Conclusion: Infection is an important inducing cause of AKI in ACLF patients. The underlying liver disease and the severity of infection are significantly related to the infection-induced AKI in ACLF patients, and once AKI occurs after infection, the mortality rate of the patients is significantly increased.

[Effects of clinical medicine on salt sensitivity hypertension: a systematic review and meta-analysis].

Objective: To assess the effects of clinical medicine on salt sensitive hypertension. Methods: The PubMed, EMBASE, Cochrane Library, CBM, WanFang Data, VIP and CNKI databases were searched to collect randomized controlled trials (RCTs) on clinical medicine in treating salt sensitive hypertension from inception to December 2018. Two reviewers independently screened the literature, extracted data, and another investigator assessed the risk of bias included in the study. Then meta-analysis was performed using RevMan 5.3 software. Results: A total of 16 RCTs studies involving 1 355 patients were included. Meta-analysis showed that angiotensin-converting enzyme inhibitors (ACEIs) combined with diuretics could effectively reduce 24 h systolic blood pressure variability [mean difference (MD)=4.45, 95%CI: 3.47-5.43, P<0.001] and 24 h diastolic blood pressure variability (MD=3.71, 95%CI:2.83-4.59, P<0.001) in salt-sensitive hypertension patients. Angiotensin Ⅱ receptor antagonists (ARBs) combined with diuretics had no antihypertensive effect on salt-sensitive hypertension patients. Indapamide alone can reduce systolic blood pressure (MD=-14.70, 95%CI:-18.57--10.83, P<0.001) and diastolic blood pressure (MD=-8.73, 95%CI:-11.57--5.89, P<0.001). The use of ACEIs alone in salt-sensitive hypertension patients can not reduce systolic pressure (MD=2.20, 95%CI:-1.48-5.88, P=0.240) and diastolic pressure (MD=2.95, 95%CI: 1.37~4.54, P<0.001). Amlodipine combined with metformin had therapeutic effect on salt-sensitive hypertension (RR=1.23, 95%CI: 1.14~1.33, P<0.001). Conclusions: ACEIs combined with diuretics can effectively reduce blood pressure variability in salt-sensitive hypertensive patients. The use of amlodipine in combination with metformin and indapamide alone have antihypertensive effect in salt-sensitive hypertensive patients.

Effects of various cryoprotectants on the quality of frozen-thawed immature bovine (Qinchuan cattle) calf testicular tissue.

To investigate the effects of different concentrations of various cryoprotectants (CPs) on the cell viability as well as expression of spermatogenesis-related genes, such as CREM, Stra8 and HSP70-2 in frozen-thawed bovine calf testicular tissue, immature bovine (Qinchuan cattle) calf testicular tissue was collected and cryopreserved in the cryomedia containing different concentrations (5%, 10%, 15% and 20%) of the following three CPs: glycerol, ethylene glycol (EG) and dimethyl sulphoxide (DMSO) respectively. After 1 month cryopreservation in liquid nitrogen, cell viability was evaluated using Trypan blue exclusion under a bright-field microscope. The mRNA expression of the three genes was also evaluated using qRT-PCR. The results indicated that different concentrations of glycerol, EG and DMSO in cryomedia during cryopreservation could protect bovine calf testicular tissue in various ways to avoid freezing or cryopreservation-induced expression changes in spermatogenesis-related genes. The highest cell viability and the three spermatogenesis-related genes (CREM, Stra8 and HSP70-2) expression level came from the cryomedia containing glycerol, EG and DMSO at 10% concentration respectively (p < .05). Meanwhile, compared with the other CPs, the frozen-thawed bovine calf testicular tissue treated with 10% DMSO exhibited the highest cell viability and mRNA expression level of the spermatogenesis-related genes (CREM, Stra8 and HSP70-2).

Effects of iodine methionine on boar sperm quality during liquid storage at 17°C.

Microbial environment is one of the important factors that affect the quality of preserved semen. Iodine methionine (IM), participating in the production and activation of metabolic enzymes, is a new type of amino acid chelate. To date, there has been no report to evaluate the effects of IM on boar semen preservation at 17°C. This study was designed to investigate the effects of IM on boar sperm quality and reproductive performance during liquid storage at 17°C and its antibacterial effect. Semen samples collected from six Yorkshire boars were diluted with basic liquid containing different concentrations of IM (0, 20, 40, 80, 160 and 320 µM). Subsequently, sperm motility, plasma membrane integrity and acrosome integrity were determined. After 6 days of preservation, the difference in microbial composition between control group and 80 µM IM group was compared using 16S rDNA sequencing, and the effects of IM on reproductive performance were also compared and analysed between the two groups. The results demonstrated that 20, 40 and 80 µM IM improved boar sperm motility, plasma membrane integrity and acrosome integrity. 80 µM IM was the optimum concentration. Conversely, 160 and 320 µM IM resulted in deleterious consequences to boar sperm quality compared to the control group and other treatment groups (p < .05). After 6 days of preservation, sperm motility, plasma membrane integrity and acrosome integrity were 56.0%, 51.8% and 59.4%, respectively. There was no significant difference in non-return rate between the two groups (p > .05). But the litter size of 80 µM IM group was significantly higher than that of control group (p < .05). 80 µM IM inhibited proliferation of the phylum Proteobacteria and the genus Staphylococcus as well as Pseudomonas (p < .05). Further studies are required to understand the antibacterial mechanism of IM in liquid-preserved boar semen.

Characterization of hemocyanin from the mud crab Scylla paramamosain and its expression analysis in different tissues, at various stages, and under Vibrio parahaemolyticus infection.

Hemocyanin is an important respiratory protein in many arthropod and mollusk species. Here, four cDNAs (SpHc1, SpHc2, SpHc3, and SpHc4), encoding distinct hemocyanin subunits from Scylla paramamosain were cloned using EST analyses and the rapid amplification of cDNA ends. The four full-length cDNA fragments (SpHc1-4) were 2281, 2002, 2184, and 2069 bp, respectively, and they encoded four putative proteins (570-676 amino acids) with a molecular mass of ~65.0-76.8 kDa. Quantitative real-time PCR analyses revealed that the four genes were mainly expressed in the hepatopancreas, testis, and hemocytes. SpHc mRNA expression during continuous developmental stages in zoeal phases (Z1, Z2, Z3, Z4, and Z5), megalopa, and juvenile crab I stages were also detected. The expression levels of SpHc3 and SpHc4 were higher than that of SpHc1 and SpHc2 during the first six stages, and they sharply declined during the juvenile stage. After infection with Vibrio parahaemolyticus, the temporal expression of both the four SpHc mRNAs in the megalopa stage rapidly declined during the first 3 h, followed by upregulation and peak expression at 12 h after the challenge. The expression levels of the four SpHc subunits were upregulated at 48 h after the challenge, and were then gradually downregulated. These findings suggest that hemocyanin may potentially be involved in the crab immune response, and that the role of the four subunits may differ in different tissues and during various developmental stages.

Protective effects of ascorbic acid and vitamin E on antioxidant enzyme activity of freeze-thawed semen of Qinchuan bulls.

The aim of this study was to determine the protective effects of the combination of ascorbic acid (Vc) and vitamin E (VE) on antioxidant enzyme activity, sperm motility, viability, and acrosome integrity of Qinchuan bulls after freeze-thaw. In this study, we determined the effects of Vc and VE on the activity of the antioxidant enzyme defense system comprising glutathione peroxidase (GSH-Px), glutathione reductase (GR), catalase (CAT), and superoxide dismutase (SOD). The combination of Vc and VE had protective effects on sperm motility and viability. With respect to acrosome integrity and the activity of GR and SOD, differences were observed between the experimental groups with added Vc (7 mg/mL) and VE (0.12 IU/mL) and the control group. The activity of GSH-Px in the experimental group (1400 IU/mL Vc and 0.12 IU/mL VE) was not different (P > 0.05) compared with that in the control group, while the activity of CAT showed a significant difference between the 2 groups (P < 0.05). Therefore, we inferred that the combination of Vc (1400 IU/mL) and VE (0.12 IU/mL) protected the sperm quality in the freeze-thaw process.

Effects of bovine serum albumin on boar sperm quality during liquid storage at 17°C.

This study aimed to investigate the effects of bovine serum albumin (BSA) on boar sperm quality during liquid storage at 17°C. Boar semen samples were collected and diluted with Modena containing different concentrations (0, 1, 2, 3, 4, 5 and 6 g/l) of BSA, and sperm motility, plasma membrane integrity, acrosome integrity, total antioxidative capacity (T-AOC) activity and malondialdehyde (MDA) content were measured and analysed. The results showed that Modena supplemented with 3, 4 and 5 g/l BSA could improve boar sperm motility, effective survival time and plasma membrane integrity (p < 0.05), decrease MDA content (p < 0.05), while no statistical difference was observed for sperm acrosome integrity and T-AOC activity among these three groups (p > 0.05). The semen sample diluted with Modena containing 4 g/l BSA could achieve optimum effect, and sperm survival time was 7.5 days. After 7 days preservation, sperm motility, plasma membrane integrity and acrosome integrity were 54%, 49% and 78%, respectively. T-AOC activity and MDA content were 1.03 U/ml and 17.5 nmol/ml, respectively. In conclusion, Modena supplemented with BSA reduced the oxidative stress and improved the sperm quality of boar semen during liquid storage at 17°C, and 4 g/l BSA was the optimum concentration. Further studies are required to obtain more concrete results on the determination of antioxidant capacities of BSA in liquid preserved boar semen.

293FT is a highly suitable mammalian cell line for the in vitro enzymatic activity analysis of typical P450 proteins.

Mammalian cells have been widely used for the in vitro evaluation of the functional effect of allelic variants of cytochrome P450 (CYP). The aim of this study was to determine the most suitable mammalian cell line for the in vitro drug metabolism analysis of CYP variants. Three reported cell lines (COS-7, HepG2, 293T) and one fast-growing variant of the 293 cell line 293FT were transfected with vectors expressing green fluorescent protein or typical variants of CYP2C9, CYP2C19 or CYP2D6 to investigate the protein expression levels and the catalytic activity of expressed CYP allelic variants. The transfected 293FT cells had the highest protein expression level and exhibited the highest enzymatic activity, while HepG2 cells showed the lowest activity among the four tested cell lines. Simultaneously, 293FT cells still maintained the similar relative enzymatic ratio among three typical CYP2C9 variants to that of the commonly used COS-7 cells. In addition, 293FT cells could also be used for the in vitro functional evaluation of two other typical P450 proteins, CYP2C19 and CYP2D6. Therefore, the 293FT cell line is more suitable for the in vitro enzymatic activity analysis of typical P450 proteins than any other reported mammalian cell lines.

Molecular characterization of thioredoxin-1 and thioredoxin reductase activity in mud crab Scylla paramamosain.

The thioredoxin (Trx) system consists of thioredoxin reductase (TrxR), Trx, and nicotinamide adenine dinucleotide phosphate (NADPH). TrxR is an NADPH-dependent oxidoreductase. Trx is a ubiquitous small protein with a redox-active disulfide bridge that plays important regulatory roles in some vital metabolic reactions. In this study, a cDNA sequence (SpTrx1) showing high identity to the first Trx gene was isolated from a hepatopancreas cDNA library of the mud crab Scylla paramamosain. The full-length cDNA of SpTrx1 consisted of 672 bp and contained a complete open reading frame of 318 bp encoding a polypeptide of 105 amino acids. Quantitative real-time polymerase chain reaction analysis revealed that SpTrx1 expression was ubiquitous in various organs of S. paramamosain, including the gill, muscle, heart, hemolymph, testis, and hepatopancreas. SpTrx1 expression was upregulated significantly after Vibrio parahaemolyticus challenge: it obviously rose at 48 h and reached the highest level at 72 h. Furthermore, TrxR activity was detected in the gill, heart, muscle, hemolymph, and hepatopancreas. The relative TrxR activity in different tissues after V. parahaemolyticus injection had the same tendency in each tissue (P < 0.01) as SpTrx1 expression. The TrxR activity increased 2 h after injection, peaked at 8 h, slowly decreased from 12 to 24 h, and returned to normal levels at 48 h. The consistency of the expression between the Trx transcript and TrxR activity demonstrated that Trx was closely related to TrxR in the Trx system in S. paramamosain, suggesting that it may participate in the immune system of mud crabs.

Lactobacillus coryniformis subsp. torquens inhibits bone loss in obese mice via modification of the gut microbiota.

High-fat diet (HFD)-induced obesity results in bone loss associated with an imbalanced gut microbiota and altered immune status. Probiotics are live microorganisms that are beneficial to the host and are important in maintaining bone health and gut homeostasis. In this study, the probiotic Lactobacillus coryniformis subsp. torquens (T3L) was isolated from traditional yak milk cheese produced in Lhasa and showed distinct acid and bile salt resistance as potential probiotics. Our data indicated that T3L not only reversed HFD-induced gut dysbiosis, as indicated by decreased Firmicutes-to-Bacteroidetes ratios but also reduced bone loss. The anti-obesity, microbiome-modulating, and bone-protective effects were transmissible via horizontal faeces transfer from T3L-treated mice to HFD-fed mice. The protective effects of T3L on bone mass were associated with regulatory T (Treg) cell-mediated inhibition of osteoclast differentiation. Our data indicate that T3L is a regulator of the gut microbiota and bone homeostasis in an animal model.