Changes in brain susceptibility in Wilson's disease patients: a quantitative susceptibility mapping study.

AIM: To assess changes in the susceptibility of the caudate nucleus (CN), putamen, and globus pallidus (GP) in patients with neurological and hepatic Wilson's disease (WD) by quantitative susceptibility mapping (QSM). MATERIAL AND METHODS: The brain MRI images of 33 patients diagnosed with WD and 20 age-matched controls were analysed retrospectively. All participants underwent brain T1-weighted, T2-weighted, and QSM imaging using a 1.5 T magnetic resonance imaging (MRI) machine. QSM maps were evaluated with the STISuite toolbox. The quantitative susceptibility levels of the CN, putamen, and GP were analysed using region of interest analysis on QSM maps. Differences among neurological WD patients, hepatic patients, and controls were determined. RESULTS: Susceptibility levels were significantly higher for all examined structures (CN, putamen and GP) in patients with neurological WD compared with controls (all p<0.05) and hepatic WD patients (all p<0.05). No statistically significant differences were found in susceptibility levels between patients with hepatic WD and controls (all p>0.05). CONCLUSION: The QSM technique is a valuable tool for detecting changes in brain susceptibility in WD patients, indicating abnormal metal deposition. Notably, the current findings suggest that neurological WD patients exhibit more severe susceptibility changes compared with hepatic WD patients. Therefore, QSM can be utilised as a complementary method to detect brain injury in WD patients.

[Clinical trials on intrathecal pemetrexed treated leptomeningeal metastases from solid tumors].

Objective: To investigate the feasibility, safety and efficacy of intrathecal pemetrexed (IP) treated for patients with leptomeningeal metastases (LM) from solid tumors. Methods: Forty-seven patients receiving pemetrexed intrathecal chemotherapy in the First Hospital of Jilin University from 2017 to 2018 were selected. The study of pemetrexed intrathecal chemotherapy adopted the classical dose-climbing model and included 13 patients with meningeal metastasis of non-small cell lung cancer who had relapsed and refractory after multiple previous treatments including intrathecal chemotherapy. Based on the dose climbing study, 34 patients with meningeal metastasis of solid tumor who did not receive intrathecal chemotherapy were enrolled in a clinical study using pemetrexed as the first-line intrathecal chemotherapy combined with radiotherapy. Kaplan-Meier method and Log rank test were used for survival analysis, and Cox regression model was used for influencing factor analysis. Results: The dose climbing study showed that the maximum tolerated dose of pemetrexed intrathecal chemotherapy was 10 mg per single dose, and the recommended dosing regimen was 10 mg once or twice a week. The incidence of adverse reactions was 10 cases, including hematological adverse reactions (7 cases), transaminase elevation (2 cases), nerve root reactions (5 cases), fatigue and weight loss (1 case). The incidence of serious adverse reactions was 4, including grade 4-5 poor hematology (2 cases), grade 4 nerve root irritation (2 cases), and grade 4 elevated aminotransferase (1 case). In the dose climbing study, 4 patients were effectively treated and 7 were disease controlled. The survival time was ranged from 0.3 to 14.0 months and a median survival time was 3.8 months. The clinical study of pemetrexed intrathecal chemotherapy combined with radiotherapy showed that the treatment mode of 10 mg pemetrexed intrathecal chemotherapy once a week combined with synchronous involved area radiotherapy 40 Gy/4 weeks had a high safety and reactivity. The incidence of major adverse reactions was 52.9% (18/34), including hematologic adverse reactions (13 cases), transaminase elevation (10 cases), and nerve root reactions (4 cases). In study 2, the response rate was 67.6% (23/34), the disease control rate was 73.5% (25/34), the overall survival time was ranged from 0.3 to 16.6 months, the median survival time was 5.5 months, and the 1-year survival rate was 21.6%. Clinical response, improvement of neurological dysfunction, completion of concurrent therapy and subsequent systemic therapy were associated with the overall survival (all P<0.05). Conclusions: Pemetrexed is suitable for the intrathecal chemotherapy with a high safety and efficacy. The recommended administration regimen was IP at 10 mg on the schedule of once or twice per week. Hematological toxicity is the main factor affecting the implementation of IP. Vitamin supplement can effectively control the occurrence of hematological toxicity.

[Molluscicidal activity of the Bacillus velezensisis Y6 strain against Oncomelania hupensis and its preliminary mechanisms of molluscicidal actions].

OBJECTIVE: To assess the molluscicidal activity of the of Bacillus Y6 strain against Oncomelania hupensis in laboratory, and to preliminarily investigate its mechanisms of molluscicidal actions. METHODS: Biological identification of the Y6 strain was performed based on analysis of its morphological and physiochemical features and homology analysis of the 16S rDNA gene sequence. Bacillus Y6 suspensions were formulated at concentrations of 0.005, 0.010 g/mL and 0.015 g/mL, and the molluscicidal activity of Bacillus Y6 suspensions against O. hupensis was tested in laboratory using the immersion method. In addition, the Bacillus Y6 content and glycogen content were detected in O. hupensis following exposure to Bacillus Y6 suspensions to preliminarily explore the molluscicidal mechanism of the Bacillus Y6 strain against O. hupensis. RESULTS: The colony of the Bacillus Y6 strain appeared non-transparent milky white, and mycoderma was produced on the surface of the nutrient agar liquid medium. The Y6 stain was Gram positive and rod-shaped, and the endospore was located at the center of the Bacillus Y6 strain and appeared an achromatic, transparent and refractive body, which was encapsulated by the Y6 strain. The Y6 strain was positive for the lecithinase test, and the 16S rDNA gene sequence showed a 100% homology with those of multiple B. velezensisis strains, B. amyloliquefaciens and B. subtilis. The Y6 strain was therefore identified as B. velezensisis. Following immersion in the Bacillus Y6 suspensions at concentrations of 0.005, 0.010 g/mL and 0.015 g/mL for 24, 48 h and 72 h, the mortality rates of Oncomelania snails were 28.3%, 31.7% and 81.6%, 43.3%, 58.3% and 93.3%, and 63.3%, 78.3% and 98.3%, respectively. The molluscicidal activity of the Bacillus Y6 suspensions increased with the suspension concentration and duration of immersion. Microscopy and colony counting revealed the highest Y6 content in dead snails and the lowest in living snails following immersion in Bacillus Y6 suspensions, and the mean glycogen contents were (0.68 ± 0.06), (1.09 ± 0.16) µg/mg and (2.56 ± 0.32) µg/mg in the soft tissues of dead, dying and living snails following immersion in Bacillus Y6 suspensions (F = 59.519, P < 0.05), and the mean glycogen content was significantly higher in living snails than in dead (t = 14.073, P < 0.05) and dying snails (t = 10.027, P < 0.05), while the mean glycogen content was significantly higher in dying snails than in dead snails (t = 5.983, P < 0.05). CONCLUSIONS: The B. velezensisis Y6 strain shows a high molluscicidal activity against O. hupensis snails, and its invasion may cause glycogen metabolism disorders, leading to snail death.

[Intervention effect of biofeedback combined with pelvic floor muscle exercise on low anterior resection syndrome in patients with low anus-preserving rectal cancer].

Objective: To explore the effect of biofeedback training combined with pelvic floor muscle exercise on the recovery of anorectal function in patients with middle and low rectal cancer undergoing sphincter-preserving surgery, and to find the best way to prevent low anterior resection syndrome. Methods: A single-center prospective randomized controlled study was conducted. One hundred and nine patients with mid-low rectal cancer in Sun Yat-sen University Cancer Centre from June 2015 to December 2016 were enrolled in the study, who were going to undergo sphincter-preserving surgery or preventive ostomy after preoperative chemoradiotherapy. They were divided into three groups: blank control group, pelvic floor muscle exercise group and biofeedback training combined with pelvic floor muscle exercise group. Intervention and follow-up were conducted for 16 months. High-resolution anorectal manometry was used to measure the objective anorectal pressure and sensory index of patients, and the Chinese version of MSKCC Intestinal Function Questionnaire was used to evaluate the intestinal function of patients. The differences of objective anorectal manometry index and subjective intestinal function between the three groups were compared, and the occurrence of low anterior rectal resection syndrome was evaluated. Results: At the end of the intervention, the total scores of anal resting pressure, rectal resting pressure, anal maximum systolic pressure, anal maximum systolic time, initial rectal volume, rectal fecal sensory capacity, rectal maximum tolerance capacity, rectal compliance, anorectal hypertensive zone and total score of intestinal function in the biofeedback training combined with pelvic floor muscle exercise group were (44.83±9.01) mmHg, (4.31±1.75) mmHg, (130.46±10.00) mmHg, (19.94±4.30) s, (32.71±5.00) ml, (74.26±8.30) ml, (188.4±12.68) ml, (5.69±1.18) ml/kPa, (3.31±0.96) cm and (68.09±6.38) points respectively. The main effects of the changes of five indices, including anal resting pressure, rectal resting pressure, anal maximum systolic pressure, anal maximum systolic time and anal high pressure zone, were time. Significant differences were found in initial rectal capacity, sensory capacity of rectal defecation, maximum tolerance capacity of rectum, rectal compliance and total score of intestinal function in every time point of measurement in the biofeedback training group combined with pelvic floor muscle exercise group. They were significantly higher than those in the blank control group (P<0.05); the score of the biofeedback training group combined with pelvic floor muscle exercise group at one month after operation, perioperative period and 3 months after operation were significantly higher than those in pelvic floor muscle exercise group (P<0.05). Biofeedback training combined with pelvic floor muscle exercise reduced the incidence of low anterior resection syndrome of rectum (P<0.05). Conclusion: Biofeedback training combined with pelvic floor muscle exercise can significantly improve the sensory indicators of patients with mid-low rectal cancer, promote the recovery of intestinal function, and alleviate low anterior resection syndrome of rectal cancer patients, which is worthy of popularization and application.

[The localization diagnosis of patients with adrenocorticotropic hormone-dependent Cushing's syndrome in adolescence].

Objective: To elucidate the clinical characteristics and localization diagnosis of patients with adrenocorticotropic hormone (ACTH)-dependent Cushing's syndrome (CS) in adolescence. Methods: The clinical data, laboratory examination and localization diagnosis from 35 patients aged less than 18 years old with adolescent CS who were treated at Peking Union Medical College Hospital between January 1990 and March 2012 were analyzed. Results: There were 29 cases of Cushing's disease (CD) and 6 cases of ectopic ACTH syndrome (EAS). Compared to patients with EAS, those with CD were older at diagnosis[(15.2±2.7) vs (12.8±4.4) years], and had longer disease course[(1.9±1.5) vs (0.7±0.3) years]and higher serum potassium[(3.8±0.6) vs (2.5±0.7) mmol/L], however the plasma ACTH level[(15.4±14.9) vs (42.5±22.7) pmol/L]was lower (all P<0.05). If the cut-off of the ratio of 24-hour urine free cortisol (24 h UFC) after low-dose dexamethasone suppression test (LDDST) to before LDDST was 0.65, the sensibility to diagnose CD was 70.8%, and the specificity was 100%. If the cut-off of the 24 h UFC ratio after high-dose dexamethasone suppression test (HDDST) to before HDDST was 0.54, the sensibility to diagnose CD was 91.7%, and the specificity was 100%. If the cut-off of the plasma ACTH ratio of inferior petrosal vein[bilateral inferior petrosal sinus sampling (BIPSS)]to peripheral vein was 2, only 6 CD patients (6/8) met it. Conclusion: The study suggested that HDDST was more meaningful in the localization diagnosis of patients with ACTH-dependent CS in adolescence.

First Report of Leaf Spot Disease Caused by Glomerella magna on Lobelia chinensis in China.

Lobelia chinensis is a perennial herbaceous plant in the family Campanulaceae that is native to China, where it grows well in moist to wet soils. It is commonly used as a Chinese herbal medicine. In May 2012, symptoms of leaf spot were observed on leaves of L. chinensis in Nanning, Guangxi Zhuang Autonomous Region, China. The leaf lesions began as small, water-soaked, pale greenish to grayish spots, which enlarged to gray to pale yellowish spots, 4 to 6 mm in diameter. At later stages, numerous acervuli appeared on the lesions. Acervuli were mostly epiphyllous, and 40 to 196 µm in diameter. On potato dextrose agar (PDA), a fungus was consistently recovered from symptomatic leaf samples, with a 93% isolation rate from 60 leaf pieces that were surface sterilized in 75% ethanol for 30 s and then in 0.1% mercuric chloride for 45 s. Three single-spore isolates were used to evaluate cultural and morphological characteristics of the pathogen. Setae were two to three septate, dark brown at the base, acicular, and up to 90 µm long. Conidia were long oblong-elliptical, guttulate, hyaline, and 11 to 20 × 4.1 to 6.3 µm (mean 15.2 × 5.1 µm). These morphological characteristics of the fungus were consistent with the description of Colletotrichum magna (teleomorph Glomerella magna Jenkins & Winstead) (1). The rDNA internal transcribed spacer (ITS) region of one isolate, LC-1, was sequenced (GenBank Accession No. KC815123), and it showed 100% identity to G. magna, GenBank HM163187.1, an isolate from Brazil cultured from papaya (2). Although KC815123 was identified as G. magna, it shows 99% identity to GenBank sequences from isolates of C. magna, and more research is needed to elucidate the relationships between these taxa, especially with consideration to host specificity. Pathogenicity tests were performed with each of the three isolates by spraying conidial suspensions (1 × 106 conidia/ml) containing 0.1% Tween 20 onto the surfaces of leaves of 30-day-old and 6- to 8-cm-high plants. For each isolate, 30 leaves from five replicate plants were treated. Control plants were treated with sterilized water containing 0.1% Tween 20. All plants were incubated for 36 h at 25°C and 90% relative humidity in an artificial climate chamber, and then moved into a greenhouse. Seven days after inoculation, gray spots typical of field symptoms were observed on all inoculated leaves, but no symptoms were seen on water-treated control plants. Koch's postulates were fulfilled by reisolation of G. magna from diseased leaves. To our knowledge, this is the first report of G. magna infecting L. chinensis worldwide. References: (1) M. Z. Du et al. Mycologia 97:641, 2005. (2) R. J. Nascimento et al. Plant Dis. 94:1506, 2010.

First Report of Leaf Spot Caused by Corynespora cassiicola on Baphicacanthus cusia in China.

Baphicacanthus cusia is a perennial herbaceous plant in the family Acanthaceae that is native to China, where it grows in warm temperate mountainous or hilly regions. It is commonly used as a Chinese herbal medicine. In March 2012, symptoms of leaf spot were observed on leaves of B. cusia in Long'an County, Guangxi, China, where this plant is extensively cultivated. Symptoms were initially small brown dots which developed into irregular to circular leaf spots. These spots enlarged and overlapped, extending until the 7- to 9-cm-long and 3- to 4-cm-wide leaves withered entirely, mostly within 2 months. On potato dextrose agar (PDA), the same fungus was cultured from 92% of 75 symptomatic leaf samples that had been surface sterilized in a 45-second dip in 0.1% mercuric chloride. Fungal structures were observed on diseased leaves: conidiophores (85 to 460 × 4 to 8 µm) were erect, brown, single or in clusters, and conidia (36 to 90 × 5 to 16 µm) were single or in chains of two to four, brown, cylindrical or obclavate, straight or slightly curved, with 3 to 18 pseudosepta and a conspicuous hilum. Three single-spore isolates were identified as Corynespora cassiicola (Berk & Curt.) Wei based on morphological and cultural characteristics (1). The rDNA internal transcribed spacer (ITS) region of one isolate, ZY-1, was sequenced (GenBank Accession No. JX908713), and it showed 100% identity to C. cassiicola, GenBank FJ852716, an isolate from Micronesia cultured from Ipomoea batatas (2). Pathogenicity tests were performed with each of the three isolates by spraying conidial suspensions (5 × 104 conidia/ml) containing 0.1% Tween 20 onto the surfaces of leaves of 60-day-old, 20-cm tall plants. For each isolate, 30 leaves from five replicate plants were treated. Control plants were treated with sterilized water containing 0.1% Tween 20. All plants were incubated for 36 h at 25°C and 90% relative humidity in an artificial climate chamber, and then moved into a greenhouse. Seven days after inoculation, dark brown spots typical of field symptoms were observed on all inoculated leaves, but no symptoms were seen on water-treated control plants. Koch's postulates were fulfilled by reisolation of C. cassiicola from diseased leaves. To our knowledge, this is the first report of C. cassiicola infecting B. cusia worldwide. References: (1) M. B. Ellis. Dematiaceous Hyphomycetes. Commonwealth Mycological Institute: Kew, Surrey, England, 1971. (2) L. J. Dixon et al. Phytopathology 99:1015, 2009.

Alternative splicing of KCNQ2 potassium channel transcripts contributes to the functional diversity of M-currents.

The region of alternative splicing in the KCNQ2 potassium channel gene was determined by RNase protection analysis of KCNQ2 mRNA transcripts. Systematic analysis of KCNQ2 alternative splice variant expression in rat superior cervical ganglia revealed multiple variant isoforms. One class of KCNQ2 splice variants, those that contained exon 15a, was found to have significantly different kinetics to those of the other isoforms. These transcripts encoded channel subunits that, when co-expressed with the KCNQ3 subunit, activated and deactivated approximately 2.5 times more slowly than other isoforms. Deletion of exon 15a in these isoforms produced a reversion to the faster kinetics. Comparison of the kinetic properties of the cloned channel splice variants with those of the native M-current suggests that alternative splicing of the KCNQ2 gene may contribute to the variation in M-current kinetics seen in vivo.

GFP-centrin as a marker for centriole dynamics in living cells.

A long-standing puzzle in cell biology is the question of how cells generate one and only one new centrosome in each cell cycle and what is the role of the centriole pair in this process. In this study, the introduction of GFP-centrin into cultured cells allows direct visualization of centriole behavior in living cells and in real time. Using this method, centriole dynamics can be observed throughout the cell cycle and following a variety of experimental treatments. Our studies demonstrate that the biogenesis of new centrioles from individual members of a preexisting centriole pair is asynchronous: the older centriole initiates assembly of a new daughter centriole before the younger centriole initiates assembly of its daughter.

Depletion of intracellular Ca2+ by caffeine and ryanodine induces apoptosis of chinese hamster ovary cells transfected with ryanodine receptor.

Recent studies have suggested a central role for Ca(2+) in the signaling pathway of apoptosis and certain anti-apoptotic effects of Bcl-2 family of proteins have been attributed to changes in intracellular Ca(2+) homeostasis. Here we report that depletion of Ca(2+) from endoplasmic reticulum (ER) leads to apoptosis in Chinese hamster ovary cells. Stable expression of ryanodine receptor (RyR) in these cells enables rapid and reversible changes of both cytosolic Ca(2+) and ER Ca(2+) content via activation of the RyR/Ca(2+) release channel by caffeine and ryanodine. Sustained depletion of the ER Ca(2+) store leads to apoptosis in Chinese hamster ovary cells, whereas co-expression of Bcl-xL and RyR in these cells prevents apoptotic cell death but not necrotic cell death. The anti-apoptotic effect of Bcl-xL does not correlate with changes in either the Ca(2+) release process from the ER or the capacitative Ca(2+) entry through the plasma membrane. The data suggest that Bcl-xL likely prevents apoptosis of cells at a stage downstream of ER Ca(2+) release and capacitative Ca(2+) entry.

[Effects of oral calcium supplementation on blood pressure in population].

OBJECTIVE: To assess the effects of oral intake of calcium lactate tablet on blood pressure and serum levels of sodium, potassium, calcium and phosphors. METHODS: In a rural area with natural population with higher average blood pressure and prevalence of hypertension, and with high salt and low calcium intake in diet in Yu County, Shanxi Province, 112 volunteers aged 30 to 64 years were selected based on a mass screening for hypertension, and randomized into trial and control groups with double blinding, taking calcium lactate tablets (800 mg of calcium daily) and placebo respectively for five weeks. Ninety-eight of them complied with the medical order completely, 39 males and 59 females, and were followed-up for systolic (SBP) and diastolic blood pressure (DBP), and their fasting venous blood specimen and 8-hour nocturnal urine specimens were collected. Twenty-four-hour diet recall was obtained for three consecutive days from all of them. RESULTS: Reduction of 4.7 mm Hg (0.62 kPa) and 2.7 mm Hg (0.36 kPa) in systolic and diastolic blood pressure, respectively in the trial group was observed, as compared with that in control one, with P-values of 0.027 and 0.074, respectively, after 35 days of treatment. The higher their baseline BP level, the greater reduction of their BP after treatment. Urinary excretion of calcium increased and serum level of phosphorus reduced after oral administration of calcium. CONCLUSION: In an area with higher prevalence of hypertension and with high salt and low calcium intake in diet, change in diet composition or supplementation with calcium could reduce their average blood pressure and prevent them from hypertension for those with mild and moderate hypertension and in high risk with higher blood pressure.

Interaction between protein kinase C and sphingomyelin/cholesterol.

Physical characteristics of binding of protein kinase C with sphingomyelin/cholesterol lipid bilayers were analysed using three complementary approaches: acrylodan fluorescence, fluorescence energy transfer and quenching of tryptophan fluorescence. It was demonstrated that sphingomyelin/cholesterol lipid membranes were available for protein kinase C binding. The intensity of the binding was dependent on the sphingomyelin content. The results of quenching of intrinsic tryptophan fluorescence showed that the enzyme molecule penetrated the sphingomyelin/cholesterol lipid bilayer to the C-16 position of labeled fatty acid probes. Our results also showed sphingomyelin itself restrains protein kinase C activity. A possible explanation for our results is that caveolae function as signaling storage devices.

Cloning of a mammalian elk potassium channel gene and EAG mRNA distribution in rat sympathetic ganglia.

1. Three new members of the EAG potassium channel gene family were identified in rat and the complete coding sequence of one of these genes (elk1) was determined by cDNA cloning. 2. The elk1 gene, when expressed in Xenopus oocytes, encodes a slowly activating and slowly deactivating potassium channel. 3. The elk1 gene is expressed in sympathetic ganglia and is also expressed in sciatic nerve. 4. Six of the seven known EAG genes were found to be expressed in rat sympathetic ganglia, suggesting an important functional role for these channels in the sympathetic nervous system.

[Quality control for huoxiang-zhengqi liquid].

The Rhizoma Atractylodis, Cortex Magnoliae Officinalis and Pericarpium Citri Reticulatae in Huoxiang-zhengqi Liquid were identified by TLC. The total contents of magnolol and honokiol were determined by HPLC to be no less than 1.9 mg/ml, with an average recovery of 104.4% and 103.3%, RSD 1.23% and 0.91% respectively.

Identification of two nervous system-specific members of the erg potassium channel gene family.

Two new potassium channel genes, erg2 and erg3, that are expressed in the nervous system of the rat were identified. These two genes form a small gene family with the previously described erg1 (HERG) gene. The erg2 and erg3 genes are expressed exclusively in the nervous system, in marked contrast to erg1, which is expressed in both neural and non-neural tissues. All three genes are expressed in peripheral sympathetic ganglia. The erg3 channel produces a current that has a large transient component at positive potentials, whereas the other two channels are slowly activating delayed rectifiers. Expression of the erg1 gene in the sympathetic nervous system has potential implications for the etiology of the LQT2 form of the human genetic disease long QT syndrome.

[Influence of cerebral traumatic dementia treated with acupuncture at houxi and shenmen].

Sixty-two cerebral traumatic dementia patients were randomly divided into two groups acupuncture treatment group (Group A, 32 cases) and physical treatment group (Group B, 30 cases). The changes of MMSE scores and auditory P300 were observed before and after the treatment. It were found that the indexes in group A changed significantly (P < 0.05-0.001), and the indexes in group B didn't change significantly (P > 0.1-0.4). The therapeutic effect in group A was better than that in group B(P < 0.001). The results confirmed that the cognitive function of cerebral traumatic dementia patients was increased effectively with acupuncture at Houxi and Shenmen.

[Effect of acupuncturing houxi and shenmen in treating cerebral traumatic dementia].

UNLABELLED: Sixty-two cerebral traumatic dementia patients were randomly divided into two groups: acupuncture group (32 cases) and physiotherapy group (30 cases). The changes of mini-mental status examination (MMSE) scores and auditory evoked potential P300 were observed before and after treatment. RESULTS: marked effective rate and effective rate of acupuncture group were 46.9% and 81.3% respectively, significantly higher than 10.0% and 30.0% of physiotherapy group (P < 0.01). It was found that after treatment the above-mentioned indexes in acupuncture group changed significantly (P < 0.05 - 0.001), while in physiotherapy group were not (P > 0.05). The therapeutic effect in acupuncture group was better than that in physiotherapy group (P < 0.001). The results confirmed that the cognitive function of cerebral traumatic dementia patients was enhanced effectively with acupuncturing at Houxi and Shemen.