[Effect of Chlorambucil Combined with Ibrutinib on Mantle Cell Lymphoma Cell Line Jeko-1 and Its Related Mechanism].

OBJECTIVE: To investigate the toxic effect of chlorambucil combined with ibrutinib on mantle cell lymphoma (MCL) cell line Jeko-1 and its related mechanism. METHODS: The MCL cell line Jeko-1 was incubated with different concentrations of chlorambucil or ibrutinib or the combination of the two drugs, respectively. CCK-8 assay was used to detect the proliferation of the cells, and Western blot was used to measure the protein expression levels of BCL-2, caspase-3, PI3K, AKT and P-AKT. RESULTS: After Jeko-1 cells were treated with chlorambucil (3.125, 6.25, 12.5, 25, 50 µmol/L) and ibrutinib (3.125, 6.25, 12.5, 25, 50 µmol /L) alone for 24, 48, 72h respectively, the cell proliferation was inhibited in a time- and dose-dependent manner. Moreover, the two drugs were applied in combination at low doses (single drug inhibition rate<50%), and the results showed that the combination of two drugs had a more significant inhibitory effect (all P < 0.05). Compared with the control group, the apoptosis rate of the single drug group of chlorambucil (3.125, 6.25, 12.5, 25, 50 µmol/L) and ibutinib (3.125, 6.25, 12.5, 25, 50 µmol/L) was increased in a dose-dependent manner. The combination of the two drugs at low concentrations (3.125, 6.25, 12.5 µmol/L) could significantly increase the apoptosis rate compared with the corresponding concentration of single drug groups (all P < 0.05). Compared with control group, the protein expression levels of caspase-3 in Jeko-1 cells were upregulated, while the protein expression levels of BCL-2, PI3K, and p-AKT/AKT were downregulated after treatment with chlorambucil or ibrutinib alone. The combination of the two drugs could produce a synergistic effect on the expressions of the above-mentioned proteins, and the differences between the combination group and the single drug groups were statistically significant (all P < 0.05). CONCLUSION: Chlorambucil and ibrutinib can promote the apoptosis of MCL cell line Jeko-1, and combined application of the two drugs shows a synergistic effect, the mechanism may be associated with the AKT-related signaling pathways.

Transcriptome and metabolome analyses revealed the response mechanism of pepper roots to Phytophthora capsici infection.

BACKGROUND: Phytophthora root rot caused by the oomycete Phytophthora capsici is the most devastating disease in pepper production worldwide, and current management strategies have not been effective in preventing this disease. Therefore, the use of resistant varieties was regarded as an important part of disease management of P. capsici. However, our knowledge of the molecular mechanisms underlying the defense response of pepper roots to P. capsici infection is limited. METHODS: A comprehensive transcriptome and metabolome approaches were used to dissect the molecular response of pepper to P. capsici infection in the resistant genotype A204 and the susceptible genotype A198 at 0, 24 and 48 hours post-inoculation (hpi). RESULTS: More genes and metabolites were induced at 24 hpi in A204 than A198, suggesting the prompt activation of defense responses in the resistant genotype, which can attribute two proteases, subtilisin-like protease and xylem cysteine proteinase 1, involved in pathogen recognition and signal transduction in A204. Further analysis indicated that the resistant genotype responded to P. capsici with fine regulation by the Ca2+- and salicylic acid-mediated signaling pathways, and then activation of downstream defense responses, including cell wall reinforcement and defense-related genes expression and metabolites accumulation. Among them, differentially expressed genes and differentially accumulated metabolites involved in the flavonoid biosynthesis pathways were uniquely activated in the resistant genotype A204 at 24 hpi, indicating a significant role of the flavonoid biosynthesis pathways in pepper resistance to P. capsici. CONCLUSION: The candidate transcripts may provide genetic resources that may be useful in the improvement of Phytophthora root rot-resistant characters of pepper. In addition, the model proposed in this study provides new insight into the defense response against P. capsici in pepper, and enhance our current understanding of the interaction of pepper-P. capsici.

[Effect of Chlorambucil on the Apoptotic Signaling Pathway in Lymphoma Cells].

OBJECTIVE: To study whether chlorambucil has apoptotic effect on the B cell lymphoma A20 cells and its exact mechanisms in apoptotic signaling pathway. METHODS: The experimental cells were treated with 20 µmol/L chlorambucil, the control cells were treated with PBS. Annexin V-FITC Cell Apoptosis Detection Kit was used to examine cell apoptosis. Western blot was used to detect the expressions of active caspase-3, Survivin, NF-κB and pAKT. Real-time fluorescent quantitative PCR was performed to examine the mRNA expression of Survivin. RESULTS: Compared with the control group, the proportion of FITC+/PI+ apoptotic cells and the expression of active caspase-3 (t=7.384, P=0.000) in the chlorambucil treatment group was significantly elevated. However, the expression of Survivin mRNA (t=4.384, P=0.000), protein expressions of survivin (t=12.360, P=0.000), NF-κB (t=5.462, P=0.000) and pAKT (t=7.183, P=0.000) in the chlorambucil-treated group all significantly decreased. CONCLUSION: The chlorambucil can induce the apoptosis of lymphoma cells, its mechanism may related with inhibition of PI3K/AKT signaling pathway, and expression of NF-κB and survivin.

[Mutation Rate and Clinical Characteristics of CALR, JAK2 V617F and MPL W515K Genes in Patients With Primary Thrombocythemia].

OBJECTIVE: To analyze the mutation rate and clinical characteristics of CALR, MPL W515K and JAK2 V617F genes in patients with primary thrombocythemia (PT). METHODS: Fifty-six patients with PT were selected as the research objects in our hospital. The CALR and MPL W515K gene mutations were determined by genomic DNA-PCR direct sequencing of the PCR products, and the JAK2 V617F gene mutation was detected by allele specific PCR method. RESULTS: Among the 56 patients with PT there were 14 cases of CALR gene mutation with the incidence rate of 25%, including 6 cases of type I, 5 cases of type II and 3 cases of type III. The sex, age, platelet(Plt) count, white blood cell (WBC) count and hemoglobin (Hb) level in the type I case of CALR gene mutation all were not significantly different from that in type II and III(all P>0.05); the WBC level in type III group significantly increased in comparison of type II group (P<0.05), while the sex, age, Hb and Plt levels showed no significant difference between the type III and type II groups (P>0.05). There were 3 cases of MPL W515K gene mutation with the incidence rate of 5.36%; 21 cases of JAK2 V617F gene mutation with the incidence rate of 37.50%. There were 13 cases of CALR gene mutation in negative patients with MPL W515K and JAK2 V617F (18 cases) with 72.22% incidence rate (13/18), and there was no cases of 1 or 2 gene mutations coexisted. The levels of Hb and WBC in peripheral blood of patients with CALR mutation were significantly lower than those of JAK2 V617F mutation (both P<0.05). In 56 cases, there were 3 cases of abnormal karyotype, with the incidence rate of 5.36%. The mutation rate of CALR gene in abnormal karyotypes (66.67%) was significantly higher than that of normal karyotypes (20.75%) (P<0.01). CONCLUSION: The incidence of JAK2 V617F gene mutation increases in the patients with primary thrombocythemia; CALR mutation rate is higher in the patients with negative MPL W515K and JAK2 V617F gene mutation, which may closely correlate with abnormal karyotype; the levels of peripheral Hb and WBC in PT the patients with CALR gene mutation are significantly lower than those in patients with JAK2 V617F mutation.

[Curative Effect of Decitabine Combined with IAG Regimen for Senile Patients with Myelodysplastic Syndrome (MDS) Transformed Acute Myeloid Leukemia].

OBJECTIVE: To investigate the curative effect and safety of decitabine combined with IAG regimen for treating senile MDS-transformed AML patients. METHODS: Two cases of senile MDS-transformed AML were treated with decitabine combined with IAG regimen (decitabine 25 mg/d,qd,ivgtt,d1-5,Idarubicin 10 mg/d,qd,ivgtt,d6,Ara-C 10 mg/m2,q12h, sc,d 6-19,G-CSF 300 µg,qd,ih,d6-19). The efficacy and adverse reactions were observed in these cases. RESULTS: 1 case for 2 courses and 1 case for 1 course obtained complete remission(CR). The myelosuppression and infections due to neutropenia were the most frequent adverse effects, the severe nonhematologic toxicity, such as liver and kidney and gastrointestinal reactions, were not observed in these patients. CONCLUSION: Decitabine combined with IAG regimen is an effective for treating senile MDS-transformed AML patients.

[Action Mechanism of Chlorambucil against Mantle Cell Lymphoma Cell Line Jeko-1].

OBJECTIVE: To explore the action mechanism of chlorambucil against mantle cell lymphoma cell line Jeko-1. METHODS: The effect of chlorambucil on Jeko-1 cell proliferation was measured by MTT method. The effect of chlorambucil on the apoptosis of Jeko-1 cell was detected by Hoechst staining and Annexin V-FITC dual staining. The activation of PI3K/AKT signaling pathway and the expression of BAX, BCL-2, procaspase 3, procaspase 8 and procaspase 9 were detected by Western blot. RESULTS: 0, 5, 10, 20 µmol/L chlorambucil could inhibit Jeko-1 cell proliferation at 24, 48, 72 h in a time- and dose-dependent manner. Chlorambucil of 0, 5, 10, 20 µmol/L increased the apoptotic rate of Jeko-1 cells, upregulated the expression of BAX, procaspase 3, procaspase 8, procaspase 9 and PI3K, increased the phosphorylation of AKT and down-regulated the expression of BCL-2. CONCLUSION: The chlorambucil can induce the apoptosis of mantle cell lymphoma Jeko-1 cells via blocking PI3K/AKT signaling pathway.

Prevalence of insomnia and its risk factors in older individuals: a community-based study in four cities of Hebei Province, China.

OBJECTIVE: To examine the epidemiology of sleep problems and insomnia among the community older individuals in Hebei Province, China, and to investigate the potential sociodemographic and clinical correlates and medication use. METHODS: This cross-sectional study was conducted with community adults, aged 60 years or older, who resided in four major cities in Hebei province. Basic sociodemographic and clinical data were collected and analyzed. A total of 3176 older adults (1292 men, 70.2 ± 6.8 years; 1884 women, 68.8 ± 6.7 years) were interviewed. All of the participants were interviewed with a standardized questionnaire and underwent insomnia screening. RESULTS: The prevalence of insomnia was 37.75%. The most common type of sleep disturbance was difficulty maintaining sleep, followed by difficulty initiating sleep and early morning awakening. Never smoking, experiencing the loss of a parent, a history of coronary heart disease, and depression symptoms were independent risk factors for insomnia in men. Occasional drinking was an independent protective factor against insomnia in men. Older age, depression symptoms, a history of cerebral hemorrhage, hyperlipidemia, living without a spouse, and having mild cognitive impairment were independent risk factors for insomnia in women. Only 11.1% of the sample with insomnia were taking sleeping medications regularly. CONCLUSION: Insomnia is highly prevalent among the community older population in Hebei Province. The percentage of individuals regularly taking sleeping medication is low among those with insomnia. Individuals with complaints of insomnia frequently have poor physical and mental health and may need more medical attention. Comprehensive measures that involve psychosocial and personal behaviors should be implemented to alleviate insomnia in older individuals.

[Inhibitory Effect of High Concentration Insulin on K562 Cell Proliferation and Its Mechanism].

OBJECTIVE: To investigate the inhibitory effect of high concentration insulin on K562 cell proliferation and its underlying mechanism. METHODS: K562 cells were treated by different concentration of insulin and/or anti-IGF-1R antibody (IGF-1R-Ab), MTT assay and flow cytometry were used to detect the K562 cells proliferation and apoptosis, respectivety; Western blot was used to measure the expression and phosphorylation level of IGE-IR, Akt, Erk1/2 in K562 cells under the different concentration of insulin. RESULTS: MTT assay showed that less than 40 mU/ml insulin could promote K562 cell proliferation, while high concentration (> 40 mU/ml) insulin has been shown to inhibit K562 cell proliferation; Flow cytometry showed that 40 mU/ml insulin suppressed K562 cell apoptosis (P < 0.05), while 200 mU/ml insulin could significantly induce K562 cell apoptosis (P < 0.01); 0.01 to 1.0 µg/ml IGF-1R-Ab has significantly enhanced the inhibitory and inducing effects of high concentration (> 40 mU/ml) of insulin on K562 cell proliferation and apoptosis respectively (r = 0.962, P < 0.001); Western blot showed that after K562 cells were treated with different concentrations of insulin ERK, and the p-ERK expression did not change significantly, after K562 cells were treated with 200 mU/ml insulin, the expression of IGF-1R and AKT also not were changed obviously, while the phosphorylation level of IGF-1R and AKT increased. CONCLUSION: High concentration (>40 mU/ml) of insulin inhibits K562 cell proliferation and induces its apoptosis, and its mechanism may be related with the binding IGF-1R by insulin, competitively inhibiting the binding of IGF-1 and IGF-1R, the blocking the transduction of PI3K/AKT signal pathway.

[Relationship between intestinal mucosal inflammation and mental disorders in patients with irritable bowel syndrome].

OBJECTIVE: To examine the relationship between inflammation and the comorbidity of mental disorders with irritable bowel syndrome (IBS) by comparing intestinal mucosa inflammatory biomarkers in patients with and without mental disorders. METHODS: A total of 43 consecutive IBS patients fulfilling the Rome III criteria and 15 volunteers serving as controls without digestive symptoms were recruited and interviewed with Composite International Diagnostic Interview (CIDI) by the well-trained staff and thus classified as with or without mental disorders. All subjects underwent colonoscopy and biopsies were acquired from the mucosa of distal ileum and colon. CD3(+) lymphocytes, mast cells, 5-HT positive cells and (indoleamine 2,3-dioxygenase) IDO positive cells were identified immunohistologically in mucosa biopsies in volunteers (n = 13), IBS patients without mental disorder (n = 24) and IBS patients with mental disorder (n = 19). RESULTS: The incidence of mental disorders in IBS patients was significantly higher than that in the volunteers (19/43 vs 2/15, P = 0.012), including 9 patients with anxiety disorders and 8 with mood disorders. (1) The number of mast cells in IBS patients with mental disorder and that in IBS patients without mental disorder has no statistical significance ((16.7 ± 3.6)/HP vs (15.4 ± 3.1)/HP in distal ileum, (12.8 ± 2.2)/HP vs (12.3 ± 2.5)/HP in sigmoid, both P > 0.05). Similar results were seen in 5-HT positive cells ((3.7 ± 0.9)/HP vs (3.4 ± 0.8)/HP in distal ileum, (6.1 ± 1.8)/HP vs (5.2 ± 1.8)/HP in sigmoid, both P > 0.05). In distal ileum, the number of CD3(+) cells in IBS patients with mental disorder has no statistical significance with that in the IBS patients without mental disorder ((62 ± 16)/HP vs (55 ± 22)/HP, P > 0.05). Similar results were seen in IDO positive cells (6(2, 8)/HP vs 2(1, 5)/HP, P > 0.05). (2) The number of IDO positive cells from distal ileum in IBS patients with anxiety disorder was significantly higher than that in the IBS patients without mental disorder (6 (4,8) vs 2 (1,5), P = 0.018). The number of mast cells from distal ileum in the IBS patients with mood disorder were significantly higher than that in those without mental disorders ((18.3 ± 3.2)/HP vs (15.4 ± 3.1)/HP, P = 0.032). CONCLUSIONS: Mental disorders in the IBS patients may be associated with intestinal mucosal inflammation. The activation of IDO may cause the comorbidity of IBS with anxiety disorder while the activation of mast cells probably leads to the comorbidity of IBS with mood disorder.

[Advancement of insulin effecting signaling pathway of leukemia cell proliferation].

Many reports have documented a role of insulin and insulin-like growth factor 1 (IGF-1) as growth factors in many cancers. The sequence and structure of insulin receptor (IR) and IGF receptor (IGF-1R) are highly similar. Both receptors are overexpressed in leukemia cells.Studies indicate that insulin can enhance the signal of the phosphoinositide 3-kinase/Akt pathways by activating IR or IGF-1R or hybrid IR/IGF-IR receptors, resulting in the proliferation of leukemia cells. High concentration of insulin may inhibit the growth of leukemia cells, the mechanism of which remains to be unclear. Inhibiting IR and IGF-IR can diminish the proliferation of leukemia cells. Therefore, the assumption of IR/IGF-1R as a potential therapeutic target in leukemia appears reasonable. This article summarizes the recent advancement associated with the signaling pathway of insulin effecting the proliferation of leukemia cells.

[Significance of sleep problems in the diagnosis of depressive and anxiety disorders].

OBJECTIVE: To explore the significance of sleep problems in the diagnosis of depressive and anxiety disorders. METHODS: A cross-sectional study was conducted through stratified convenient sampling in outpatients at our hospital from November 19, 2009 to January 19, 2010. A total of 203 outpatients (≥ 16 years old) were recruited from Department of Internal Medicine, Department of Surgery, Department of Obstetrics & Gynecology and other departments. Sleep Questionnaire was used as a screening tool for depressive and anxiety disorders. Composite international diagnostic interview (CIDI) was conducted by trained interviewers. According to diagnostic and statistical manual of mental disorders, Fourth Edition (DSM-IV) using the outcome from the logistic diagnostic procedure of CIDI as a golden standard for depressive and anxiety disorders, the validities of Sleep Questionnaire were calculated. RESULTS: According to the Sleep Questionnaire, the 12-month prevalence of any sleep problem was 31.0% (63/203). The Cronbach's α coefficient of the questionnaire was 0.764 and the area under the ROC (receiver operating characteristic) curve 0.741. With a cut-off point of 1, the sensitivity was 68.0% and the specificity 74.2% respectively. And it achieved the best composite diagnostic capacity. The positive predictive values were at least 27.0%. CONCLUSION: Sleep problem is a common complaint at a general hospital. When the duration and severity of sleep problem meet the standards of Sleep Questionnaire, a physician should be aware of possible depressive and anxiety disorders and more detailed information should be collected.

Prevalence and correlates of active and ever-smokers in metropolitan China.

This study investigated the prevalence and demographic correlates of active and ever-smokers in Beijing and Shanghai. Using a multi-stage household probability sampling method, 5201 participants aged 18-70 underwent face-to-face interviews using the Composite International Diagnostic Interview. 67.1% and 55.5% of male and 7.1% and 5.5% of female respondents were ever-smokers and active smokers respectively. Quitting was less common by proportion among those no longer married or never-married, middle-aged or working adults. After adjusting for other sociodemographic factors, the oldest age-group (>54 years) showed a significantly negative association with active smoking while those no longer married had significantly positive association with active smoking, among ever-smokers. Although the high prevalence of male smoking in Beijing and Shanghai was expected, the prevalence of female smoking was significantly higher than those found in previous surveys. This pattern of more female smoking not accompanied by an obvious decrease in male smoking defies the expectation of an orderly transition of smoking patterns and may foreshadow smoking patterns in other parts of China.

[Efficient method for extraction of high quality RNA from microtubers of Pinellia ternata in vitro].

OBJECTIVE: To extract RNA from Pinellia ternata and lay a foundation for studying the formation mechanism of P. ternata. METHOD: By modifying the method recommended by Guanidinium for extracting total RNA from plant tissues rich in phenolic and polysaccharidic compounds, a simple and convenient method for extraction of total RNA from the tubers, stems and leaves of P. ternate containing abundant polyphenols and polysaccharides was established. High concentrated p-mercaptoethanol was added in the RNA extracted buffer to remove polyphenols, phenol and chloroform were used to eliminate proteins, and isopropanol and sodium acetate were used to precipitate polysaccharides. RESULT: The A260/A230 value of RNA extracted with improved method were all over 2.0 and the values of A260/A280 were between 1.7 and 2.0. The electrophoresis bands were cleared on agarosegel and integrity of RNA was good. CONCLUSION: The results showed that RNA obtained from the tubers, stems and leaves of P. ternate with this method had high purity and quality and could be used in molecular biological research, as DDRT-PCR and reverse Northern blotting analysis directly. This method is simple, economic, stable performance, and has a good repeatability as well as is suitable for extracting total RNA of medicinal plants with high concentrations of phenolics and polysaccharides.

[Study on the factors influencing suicidal ideation among medical students in Beijing].

OBJECTIVE: To evaluate the effects of personality-related health risk factors on suicidal ideation among medical students. METHODS: 1204 medical students at first grade were selected in Beijing, using random cluster sampling method. Data were obtained through health risk behaviors questionnaire, personality diagnostic questionnaire-4 (PDQ-4) and were analyzed by logistic regression method. RESULTS: There were 12 risk factors selected from single factor analysis, including physical fight, physical abuse, physically forced to have sexual intercourse, sexual risk behaviors, tobacco and alcohol use behaviors, loneliness, bad mood, insomnia, feeling hopeless, higher PDQ-4 score and internet abuse behaviors. Data from Unconditional logistic regression showed that the main risk factors of suicide ideation were insomnia (OR = 4.98), physical abuse (OR = 4.43), sexual risk behaviors (OR = 2.63), bad mood (OR = 2.32), feeling hopeless (OR = 1.98), higher PDQ-4 score (OR = 1.09) in male students; while fighting (OR = 7.10), loneliness (OR = 4.42), physically forced to have sexual intercourse (OR = 4.19), internet abuse behaviors( OR = 1.39) in female students. CONCLUSION: Suicidal ideation was associated with various factors, with significant gender difference.

[Change of endogenous hormone around sprout tumble of Pinellia ternata under high temperature stress].

OBJECTIVE: To study the change of endogenous hormone (ABA, IAA, JA, GA3, ZR) in the leaves, petioles, tubers of Pinellia ternate around sprout tumble. It also provided some valuable information to prevent sprout tumble and increase production. METHOD: Tubers of P. ternata were cultured firstly at (23 +/- 1) degree C for certain days, and then they were coerced under (30 +/- 1 ) degree C stress in the same artificial climate boxes. The endogenous hormones in leaves, petioles and tubers during different stages of high temperature stress were determined with Enzyme-linked Immunosorbent Assays (ELISA). RESULT: After under high temperature stress, ABA content in leaves, petioles and tubers increased obviously. Similarly, JA content rose all in the leaves, petioles and tubers. But in the same conditions IAA content declined significantly in the leaves and petioles. In the tubers, IAA content also decreased, but not quickly. With the extension of high temperature coercion, the leaves, petioles, tubers, ZR content were gradually falling off. In the leaves of GA3 content rose markedly at the third day, fell down at the sixth day, but remained higher than before treatment. With the extension of the processing time, GA3 content fell off in the petioles and tubers. CONCLUSION: ABA, JA, ZT and GA3 played an important role in controlling sprout tumble of P. ternata.

[Economic evaluation of "early detection and treatment of esophageal cancer"].

BACKGROUND & OBJECTIVE: In China, esophageal cancer is the forth death cause of malignant cancers. The survival rate of esophageal cancer patients is less than 10%. Early detection and treatment of esophageal cancer is essential to improve the survival rate of these patients. This study was to evaluate the economic and social benefits of the project "early detection and treatment of esophageal cancer" through cost-benefit analysis, so as to provide evidence to extend this project. METHODS: Interview with structured questionnaires was conducted by stratified sampling among 136 esophageal cancer patients in 3 hospitals at Linzhou City, and a sampling survey was conducted among 53 patients at their homes afterwards. The 2 surveys measured the expense of "traditional treatment of esophageal cancer". Based on an intervention study of "early detection and treatment of esophageal cancer" among 936 persons at Linzhou City, cost-benefit analysis and the evaluation of economic burden of disease among esophageal cancer patients were carried out. RESULTS: The expense of "traditional treatment of esophageal cancer" was 7,183 Yuan per patient. Regarding the 936 persons who were recruited to the intervention study, the cost was 148,246 Yuan, and the benefit was 589,006 Yuan. The difference between benefit and cost of the project was 440,760 Yuan. The benefit-cost ratio was 4:1. By the project, the ratio of the expense in treating disease to the total family income in 1 year was reduced from 1.300 to 0.125. CONCLUSION: The cost of "early detection and treatment of esophageal cancer" is low, while the benefit is high, and the project can also lighten the economic burden of esophageal cancer patients.