Huperzine A attenuates nonalcoholic fatty liver disease by regulating hepatocyte senescence and apoptosis: an in vitro study.

OBJECTIVE: This study was undertaken to detect if free fatty acids (FFA) induce hepatocyte senescence in L-02 cells and if huperzine A has an anti-aging effect in fatty liver cells. METHODS: L-02 cells were treated with a FFA mixture (oleate/palmitate, at 3:0, 2:1, 1:1, 1:2 and 0:3 ratios) at different concentrations. Cell viability and fat accumulation rate were assessed by a Cell Counting Kit 8 and Nile Red staining, respectively. The mixture with the highest cell viability and fat accumulation rate was selected to continue with the following experiment. The L-02 cells were divided into five groups, including the control group, FFA group, FFA + 0.1 µmol/L huperzine A (LH) group, FFA + 1.0 µmol/L huperzine A (MH) group and FFA + 10 µmol/L huperzine A (HH) group, and were cultured for 24 h. The expression of senescence-associated ß-galactosidase (SA-ß-gal) was detected by an SA-ß-gal staining kit. The expression levels of aging genes were measured by qRT-PCR. The expression levels of apoptosis proteins were detected by a Western blot. ELISA kits were used to detect inflammatory factors and oxidative stress products. The expression of nuclear factor (NF-κB) and IκBα were detected by immunofluorescence. RESULTS: The FFA mixture (oleate/palmitate, at a 2:1 ratio) of 0.5 mmol/L had the highest cell viability and fat accumulation rate, which was preferable for establishing an in vitro fatty liver model. The expression of inflammatory factors (TNF-α and IL-6) and oxidants Malonaldehyde (MDA), 4-hydroxynonenal (HNE) and reactive oxygen species (ROS) also increased in the L-02 fatty liver cells. The expression levels of aging markers and aging genes, such as SA-ß-gal, p16, p21, p53 and pRb, increased more in the L-02 fatty liver cells than in the L-02 cells. The total levels of the apoptosis-associated proteins Bcl2, Bax, Bax/Bcl-2, CyCt and cleaved caspase 9 were also upregulated in the L-02 fatty liver cells. All of the above genes and proteins were downregulated in the huperzine A and FFA co-treatment group. In the L-02 fatty liver cells, the expression of IκBα decreased, while the expression of NF-κB increased. After the huperzine A and FFA co-treatment, the expression of IκBα increased, while the expression of NF-κB decreased. CONCLUSION: Fatty liver cells showed an obvious senescence and apoptosis phenomenon. Huperzine A suppressed hepatocyte senescence, and it might exert its anti-aging effect via the NF-κB pathway.

Effect of posture on (13)C-urea breath test in partial gastrectomy patients.

AIM: To investigate whether posture affects the accuracy of (13)C-urea breath test ((13)C-UBT) for Helicobacter pylori (H. pylori) detection in partial gastrectomy patients. METHODS: We studied 156 consecutive residual stomach patients, including 76 with H. pylori infection (infection group) and 80 without H. pylori infection (control group). H. pylori infection was confirmed if both the rapid urease test and histology were positive during gastroscopy. The two groups were divided into four subgroups according to patients' posture during the (13)C-UBT: subgroup A, sitting position; subgroup B, supine position; subgroup C, right lateral recumbent position; and subgroup D, left lateral recumbent position. Each subject underwent the following modified (13)C-UBT: 75 mg of (13)C-urea (powder) in 100 mL of citric acid solution was administered, and a mouth wash was performed immediately; breath samples were then collected at baseline and at 5-min intervals up to 30 min while the position was maintained. Seven breath samples were collected for each subject. The cutoff value was 2.0‰. RESULTS: The mean delta over baseline (DOB) values in the subgroups of the infection group were similar at 5 min (P > 0.05) and significantly higher than those in the corresponding control subgroups at all time points (P < 0.01). In the infection group, the mean DOB values in subgroup A were higher than those in other subgroups within 10 min and peaked at the 10-min point (12.4‰ ± 2.4‰). The values in subgroups B and C both reached their peaks at 15 min (B, 13.9‰ ± 1.5‰; C, 12.2‰ ± 1.7‰) and then decreased gradually until the 30-min point. In subgroup D, the value peaked at 20 min (14.7‰ ± 1.7‰). Significant differences were found between the values in subgroups D and B at both 25 min (t = 2.093, P = 0.043) and 30 min (t = 2.141, P = 0.039). At 30 min, the value in subgroup D was also significantly different from those in subgroups A and C (D vs C: t = 6.325, P = 0.000; D vs A: t = 5.912, P = 0.000). The mean DOB values of subjects with Billroth I anastomosis were higher than those of subjects with Billroth II anastomosis irrespectively of the detection time and posture (P > 0.05). CONCLUSION: Utilization of the left lateral recumbent position during the procedure and when collecting the last breath sample may improve the diagnostic accuracy of the (13)C-UBT in partial gastrectomy patients.

Clinical analysis of pharyngeal musculature and genioglossus exercising to treat obstructive sleep apnea and hypopnea syndrome.

OBJECTIVE: To evaluate the effect of pharyngeal musculature and genioglossus exercising on obstructive sleep apnea and hypopnea syndrome (OSAHS). METHODS: We conducted a non-randomized retrospective clinical trial of 75 patients with OSAHS. Fifty-four patients were managed by exercising of the pharyngeal musculature and genioglossus (exercising group). Twenty-one patients, who refused to undertake any treatment, were defined as the control group. We took the Epworth Sleepiness Scale (ESS), checked patients' polysomnography, and took 320-detector computed tomography (CT) before treatment. Six and twelve months later, we made records of apnea hypopnea index (AHI), lowest arterial oxygen saturation (LSaO2), body mass index (BMI), the shortest sagittal diameter, and transverse diameter, and the effective rates of exercising were calculated and compared with the 21 patients without any treatment (control group) at the same time. SPSS 10.0 was used to analyze the data. RESULTS: Before treatment, the ESS value was 7.67; 6 and 12 months later, the values were 3.54 and 3.25, respectively in the exercising group. AHI was decreased to 15.36 after 6 months and 13.79 after 12 months from 22.84 at the beginning. LSaO2 values were up to 81.18% after 6 months and 81.93% after 12 months from 74.05% at the beginning. There were significant differences in ESS scores, AHI, and LSaO2 between pre-treatment and post-treatment in the exercising group (P<0.05). However, there was no statistical difference in all the parameters between 6 and 12 months of exercising. The effective rates were 70.37% and 74.07% after 6- and 12-month exercising, respectively. There were significant differences between the exercising and control groups (P<0.0001). There was no statistical difference in the effective rate of the exercising group between 6 and 12 months of exercising (P>0.05). At 12 months of exercising, the compliance of the anteroposterior pharyngeal wall of the retropalatal area was lower (P<0.01) than that before treatment. There was no significant change of BMI in either group. CONCLUSIONS: Exercising pharyngeal musculature and genioglossus is a kind of non-invasive and cost-effective method to treat some OSAHS patients, especially those who are old, without surgical complications, and especially mild and moderate OSAHS patients who do not want to take surgery and continuous positive airway pressure (CPAP) treatment. In addition, exercising pharyngeal musculature and genioglossus can be considered as remedial treatment of OSAHS to surgery and other therapies.

Clinical research on liver reserve function by 13C-phenylalanine breath test in aged patients with chronic liver diseases.

BACKGROUND: The objective of this study was to investigate whether the 13C-phenylalanine breath test could be useful for the evaluation of hepatic function in elderly volunteers and patients with chronic hepatitis B and liver cirrhosis. METHODS: L-[1-13C] phenylalanine was administered orally at a dose of 100 mg to 55 elderly patients with liver cirrhosis, 30 patients with chronic hepatitis B and 38 elderly healthy subjects. The breath test was performed at 8 different time points (0, 10, 20, 30, 45, 60, 90, 120 min) to obtain the values of Delta over baseline, percentage 13CO2 exhalation rate and cumulative excretion (Cum). The relationships of the cumulative excretion with the 13C-%dose/h and blood biochemical parameters were investigated. RESULTS: The 13C-%dose/h at 20 min and 30 min combined with the cumulative excretion at 60 min and 120 min correlated with hepatic function tests, serum albumin, hemoglobin, platelet and Child-Pugh score. Prothrombin time, total and direct bilirubin were significantly increased, while serum albumin, hemoglobin and platelet, the cumulative excretion at 60 min and 120 min values decreased by degrees of intensity of the disease in Child-Pugh A, B, and C patients (P < 0.01). CONCLUSIONS: The 13C-phenylalanine breath test can be used as a non-invasive assay to evaluate hepatic function in elderly patients with liver cirrhosis. The 13C-%dose/h at 20 min, at 30 min and cumulative excretion at 60 min may be the key value for determination at a single time-point. 13C-phenylalanine breath test is safe and helpful in distinguishing different stages of hepatic dysfunction for elderly cirrhosis patients.

[Assessment of the health-related quality of life of patients with minimal hepatic encephalopathy].

OBJECTIVE: The Medical Outcome Study of 36-item Short-Form Health Survey (SF-36) is a well-validated generic questionnaire widely used to assess health-related quality of life (HRQOL), and the Chronic Liver Disease Questionnaire (CLDQ) is a specific HRQOL assessment designed for patients with liver diseases. The aim of our study is to evaluate the HRQOL based on SF-36 and CLDQ (Chinese version) in patients with chronic hepatitis B and liver cirrhosis, especially in the status of minimal hepatic encephalopathy (MHE). METHODS: The SF-36 and CLDQ were answered by 160 healthy volunteers, 20 patients with chronic hepatitis B and 106 patients with cirrhosis. HRQOL scores of the groups with different liver disease severities and with or without MHE were compared. The SF-36 includes one multi-item scale that assesses eight health categories: physical functioning, role-physical, body pain, general health, vitality, social functioning, role-emotion, and mental health. CLDQ assesses 6 categories: abdominal symptoms, fatigue, systemic symptoms, activity, emotional function and worry. RESULTS: Compared with the healthy controls, patients with chronic hepatitis B and liver cirrhosis at baseline had a lower HRQOL on all scales of the SF-36 and CLDQ (P < 0.01 for all). Increased severity of liver cirrhosis (based on the Child-Pugh score but with MHE or without) was associated with a decrease in most components, both in SF-36 and in CLDQ. However, patients with Child-Pugh B and C disease had similar HRQOL scores on both the SF-36 and CLDQ (P > 0.05), except role-physical and vitality on SF-36. There was a significant difference between patients with and without MHE on the SF-36 score (P < 0.01), and no significant difference (P > 0.05) on CLDQ scores except in abdominal symptoms. CONCLUSION: The Chinese version of SF-36 along with CLDQ are valid and reliable methods for testing MHE in patients with liver cirrhosis.

Assessment of health-related quality of life in Chinese patients with minimal hepatic encephalopathy.

AIM: To evaluate the health-related quality of life (HRQOL) based on the Chinese version of SF-36 and Chronic Liver Disease Questionnaire (CLDQ) in subjects with chronic hepatitis B, liver cirrhosis, including patients with minimal hepatic encephalopathy (MHE). METHODS: The SF-36 and CLDQ were administered to 160 healthy volunteers, 20 subjects with chronic hepatitis B and 106 patients with cirrhosis (33 cases exhibited MHE). HRQOL scores were compared among the different study groups. The SF-36 includes eight health concepts: physical functioning, role-physical, body pain, general health, vitality, social functioning, role-emotion, and mental health. Six domains of CLDQ were assessed: abdominal symptoms, fatigue, systemic symptoms, activity, emotional function and worry. RESULTS: Compared with healthy controls (96.9 +/- 4.5, 86.6 +/- 18.4, 90.1 +/- 12.5, 89.0 +/- 5.7, 87.5 +/- 4.3, 95.8 +/- 7.1, 88.5 +/- 15.9, 88.7 +/- 5.2 in SF-36 and 6.7 +/- 0.5, 6.1 +/- 0.6, 6.3 +/- 0.6, 6.5 +/- 0.5, 6.3 +/- 0.5, 6.8 +/- 0.4 in CLDQ), patients with chronic hepatitis B (86.3 +/- 11.0, 68.8 +/- 21.3, 78.9 +/- 14.4, 60.8 +/- 10.5, 70.8 +/- 8.6, 76.1 +/- 12.6, 50.0 +/- 22.9, 72.2 +/- 10.6 and 5.5 +/- 1.0, 4.5 +/- 1.0, 5.2 +/- 1.1, 5.3 +/- 0.9, 4.8 +/- 0.9, 4.9 +/- 1.0) and cirrhosis (52.8 +/- 17.4, 32.8 +/- 27.9, 61.6 +/- 18.9, 30.2 +/- 18.3, 47.9 +/- 20.1, 54.0 +/- 19.2, 28.9 +/- 26.1, 51.1 +/- 17.8 and 4.7 +/- 1.2, 3.9 +/- 1.2, 4.7 +/- 1.2, 4.7 +/- 1.3, 4.7 +/- 1.0, 4.4 +/- 1.1) had lower HRQOL on all scales of the SF-36 and CLDQ (P < 0.01 for all). Increasing severity of liver cirrhosis (based on the Child-Pugh score/presence or absence of MHE) was associated with a decrease in most components of SF-36 and CLDQ, especially SF-36. CONCLUSION: The Chinese version of SF-36 along with CLDQ is a valid and reliable method for testing MHE in patients with liver cirrhosis. Cirrhosis and MHE are associated with decreased HRQOL.

[QTL mapping of five agronomic traits in maize].

Agronomic traits have significant influence on stability and adaptability in maize production. In this investigation, using a population with 266 F2:3 families from Yuyu22 (Zong3 x 87-1), two-location field tests were conducted in Wuhan and Xiangfan in 2001, with a randomized complete block design, to characterize five agronomic traits: ear height, tassel branch number, stalk diameter, days to pollen, and days to silk. Correlation analysis of field performance indicated that ear height, tassel branch number and stalk diameter were significantly positive correlative with single-plant yield, days to pollen and days to silk were highly positive correlative with each other, and tassel branch number was significantly positive correlative with stalk diameter too. Utilizing data of field tests and molecular markers, Composite Interval Mapping (CIM) method was used to localize the quantitative trait loci of these traits and 500 times permutation test was conducted to have proper LOD threshold value. As the results, total seven QTL of ear height, nine QTL of tassel branch number, eight QTL of stalk diameter, nine QTL of days to pollen, and seven QTL of days to silk were mapped on 10 chromosomes of maize; all of these QTL distributed unevenly on chromosomes and trended to cluster together. According to analysis of this investigation, the phenotype correlations of quantitative traits may result from the correlations of QTL controlling those traits. Those will be helpful to further understand genetic basis of agronomic traits in maize.

[Comparative analyses of QTL for important agronomic traits between maize and rice].

The objective of this study was to assess syntenic relationships of quantitative trait loci (QTL) for important agronomic traits between maize and rice based on the comparative genomic map of maize and rice using two F(2:3) populations. Through the comparisons, it was observed that there were extensive conserved relationships of maize QTL affected plant height, row number, and kernels per row with rice QTL affected plant height, tillers per plant, and grains per panicle respectively. Sixteen of 45 QTL affecting five different maize traits were conserved compared with 12 of 38 QTL affecting five different rice traits, which provided some useful information for locating, isolating and cloning maize QTL by using the rice genomic data. In this study, one QTL in rice usually had two conserved QTL in maize, further supporting the hypothesis that there is a polyploidization event during maize evolution. It was interested in observing that there were QTL rich regions on chromosomes in maize and rice, where QTL affecting different traits were usually clustered. These results revealed that the QTL affected the same or similar traits in maize and rice may have the common origin. These results will be helpful to map, isolate and clone QTL in large genome crops, such as maize, by using rice genome information, as well as to understand the evolutionary forces that structured the organization of the grass genomes.

[Genetic analysis of segregation distortion of molecular markers in maize F2 population].

A genetic linkage map of maize was constructed using 150 SSR and 24 RFLP markers, with F2 population from an elite hybrid (Zong3 x 87-1). Among 174 markers, covering whole maize 10 chromosomes, 49 markers (28.1%) showed the genetic distortion (P < 0.05). Of the total segregation distortion markers, 11 markers (22.5%) deviated toward male parent, Zong3, while 12 markers (24.5%) deviated toward female parent, 87-1, besides 25 markers (51.0%) distorted to heterozygote. Only one marker distorted to both parents. Totally, 14 segregation distortion regions (SDRs) were detected among 9 different chromosomes. Four of them were located in near regions where gametophyte genes were mapped, indicating that segregation distortion may be caused by gametophyte genes partially. Two segregation distortion regions, SDR6-1 and SDR7-1, detected in this study, seemed to be new segregation distortion regions. In this paper, reasons for segregation distortion and effects of segregation distortion on genetic mapping and QTL analysis were discussed. Regarding to QTL analysis with single locus, segregation distortion would not affect QTL mapping, but regarding to analysis of digenic interactions for epistasis, the fewer distortion markers and larger size population would be needed.