[Acidophil stem cell pituitary neuroendocrine tumors/adenoma: a clinicopathological analysis of five cases].

Objective: To investigate the clinicopathological characteristics of acidophil stem cell pituitary neuroendocrine tumors (PitNET)/adenoma. Methods: Five cases of acidophil stem cell PitNET/adenoma were diagnosed between May 2022 and July 2023 at the Second Hospital of Hebei Medical University, Shijiazhuang, China. The clinicopathological features of the tumor were analyzed by using histology, immunohistochemistry, and electron microscopy. The relevant literature was reviewed. Results: There were 1 male and 4 females, aged from 23 to 69 years. Patient 3 was 55 years old at the time of diagnosis and first surgery, and relapsed 5 years later. The patients' median age was 32 years. Patients 1 and 5 showed elevated blood prolactin, with various degrees of hormonal symptoms except Patient 3, who showed only tumor compression symptoms. Imaging studies showed that all cases involved the sellar floor. The tumors of Patients 1, 2 and 5 were closely related to the cavernous sinus segment of the internal carotid artery. The tumors exhibited a diffuse growth pattern with chromophobic to slightly acidophilic cytoplasm. A few of tumor cells showed chromophobic cytoplasm. The nucleoli were conspicuous. Intranuclear inclusion bodies and variably-sized clear vacuoles were observed occasionally. Under electron microscope, marked mitochondrial abnormalities were observed, including increased mitochondria number, expanded hypertrophy, and absence of mitochondrial ridge fracture. Some mitochondrial matrices were dense, while some were vacuolated. Conclusions: Acidophil stem cell PitNET/adenoma is a rare type of pituitary adenomas/PitNETs. It often has a more clinically aggressive manner with immature cells, diffuse expression of PIT1, prolactin, and varying degrees of growth hormone expression. Because of the obvious diversity of their clinical hormone status and hormone immune expression, the diagnosis of this type tumor is still a challenge.

[Clinical study of antinib combined with radiotherapy in the treatment of third-line extensive small cell lung cancer].

Objective: To explore whether the survival benefit of the third-line extensive small-cell lung cancer (ES-SCLC) will be obtained by the combination of anlotinib and radiotherapy, and evaluate the safety of this treatment regimen. Methods: Twenty-seven patients with ES-SCLC who received third-line treatment with less than three metastatic organs at the Cancer Hospital of Xinjiang Medical University from November 2018 to July 2021 were collected and treated with radiotherapy based on anlotinib. Kaplan-Meier curve was used to estimate the overall survival (OS) and progression-free survival (PFS), descriptive statistical analysis was used to evaluate the safety, and European organisation for research and treatment of cancer quality of life questionnaire-core 30 (EORTC QLQ-C30) was used to evaluate the quality of life. Results: The follow-up cut-off date was July 1, 2021, and the follow-up time ranged from 4.8 to 31.0 months, with a median follow-up time of 10.2 months for the entire group. Among the 27 patients, 4 achieved partial remission, 17 had stable disease and 6 had progression of disease. The objective remission rate (ORR) was 14.8%, and the disease control rate (DCR) was 77.8%. Median PFS and the median OS were 5 months and 11 months, respectively. The most common adverse reactions included fatigue (33.3%, 9/27), anorexia (14.8%, 4/27), bleeding (14.8%, 4/27) and hand-foot syndrome (11.1%, 3/27). Most of them were grade 1 to grade 2, 3 cases were more than grade 3, and there was no grade 5 toxicity recorded. After radiotherapy combined with amlotinib treatment, patients showed improvement in general health, somatic functioning, social functioning, and emotional functioning (all P<0.05). Conclusion: For the third-line ES-SCLC patients, radiotherapy based on the anlotinib can significantly prolong their PFS and OS, and the adverse reactions can be tolerated.

[Effects of psychological suggestion combined with rational food restriction therapy on blood glucose and lipid metabolism and mental resilience in patients with diabetes].

Objective: To analyze the effect of psychological suggestion combined with rational food restriction therapy on blood glucose, lipid metabolism and mental resilience in patients with diabetes. Methods: Patients with diabetes admitted to the Third Hospital of Nanchang from January 2020 to August 2020 were divided into the control group and the intervention group with randomized controlled and single blind methods. The control group was treated with routine dietary guidance and health education, and the intervention group was treated with psychological suggestion combined with rational diet therapy on the basis of the control group. Both groups were treated for 3 months. Blood glucose, lipid metabolism, mental resilience and quality of life were compared between the two groups at baseline and after 3-month intervention. Differences between groups and within groups were analyzed by t test and χ2 test. Results: 100 patients in the control group and 81 patients in the intervention group completed 3-month intervention. After 3-month intervention, the levels of glycosylated hemoglobin, fasting blood glucose, 2-hour postprandial blood glucose, low-density lipoprotein cholesterol and triglyceride in both groups were lower than those before intervention. The levels of these indicators in intervention group were lower than those in control group (P<0.05). However, the levels of high-density lipoprotein cholesterol and the scores of tenacity, self-reliance, optimism, role function, emotional function, social function, physical function and cognitive function in both groups were higher than those before intervention. These indicators in intervention group were higher than those in control group (P<0.05). Conclusion: Psychological suggestion combined with rational food restriction therapy could effectively improve the glucose and lipid metabolism, mental resilience, and quality of life among patients with diabetes.

[Risk factors and chemoprophylaxis of ulcerative colitis-colorectal cancer].

Ulcerative colitis-colorectal cancer (UC-CRC) is one of the most serious complications in patients with ulcerative colitis (UC), with worse prognosis and higher mortality than sporadic colorectal cancer (CRC). Since most UC-CRC developed through the "inflammation-dysplasia-carcinoma" approach, early detection of dysplasia through identification of high-risk groups reasonable monitoring and active prevention are extremely important. However, there is no consensus on the risk factors of UC carcinogenesis and the drugs that can be used for chemoprevention currently. This article combined with relevant literature at home and abroad, reviewed the current risk factors and chemopreventive drugs for UC carcinogenesis, in order to provide reference for early prevention, early detection and early diagnosis of UC-CRC.

[Adrenal cavernous hemangioma: A case report and literature review].

A 57-year-old male patient was referred to our department with complaints of his right adrenal gland occupancy and hypertension about 6 months. When admitted to the hospital, the blood pressure was about 160/100 mmHg, and the heart rate was 110 beats/min. He was no obvious obesity, acne, abnormal mood, without weakness of limbs, acral numbness, palpitation and headache. He presented with type 2 diabetes for more than 3 years, with oral administration of metformin enteric coated tablets and subcutaneous injection of insulin glargine to control blood glucose, and satisfied with blood glucose control. Enhanced CT showed that: the right adrenal gland showed a kind of oval isodense, slightly hypodense shadow, the edge was clear, lobular change, the size was about 5.8 cm×5.4 cm, uneven density, there were nodular and strip calcification, round lipid containing area and strip low density area, and the CT value of solid part was about 34 HU. Enhanced scan showed heterogeneous nodular enhancement in the solid part of the right adrenal gland, nodular enhancement could be seen inside. The CT values of solid part in arterial phase, venous phase and delayed phase were 45 HU, 50 HU and 81 HU, respectively. Considering from the right adrenal gland, cortical cancer was more likely. No obvious abnormality was found in his endocrine examination. After adequate preoperative preparation, retroperitoneal laparoscopic adrenalectomy was performed under general anesthesia. During the operation, the 6 cm adrenal tumor was closely related to the inferior vena cava and liver, and after careful separation, the tumor was completely removed and normal adrenal tissue was preserved. The operation lasted 180 min and the blood loss was 100 mL, and the blood pressure was stable during and after the operation. There was no obvious complication. The results of pathological examinations were as follows: the size of the tumor was 7.5 cm×6.0 cm×3.5 cm, soft, with intact capsule and grayish-red cystic in section. Pathological diagnosis: (right adrenal gland) cavernous hemangioma, secondary intravascular thrombosis, old hemorrhagic infarction with calcification and ossification. After 6 months of observation, no obvious complications and tumor recurrence were found. In summary, cavernous hemangioma of adrenal gland is a rare histopathological change. Its essence is a malformed vascular mass. Blood retention is the cause of thrombosis and calcification in malformed vessels. The imaging findings were inhomogeneous enhancement of soft tissue masses, and the adrenal function examination showed no obvious abnormalities. Retroperitoneal laparoscopic surgery is feasible after adequate preoperative preparation. It is difficult to diagnose the disease preoperatively and needs to be confirmed by postoperative pathology.

Antifungal activity of cinnamaldehyde against Fusarium sambucinum involves inhibition of ergosterol biosynthesis.

AIMS: In this study, the antifungal effect of cinnamaldehyde against Fusarium sambucinum and its underlying mechanisms were determined. METHODS AND RESULTS: Minimum inhibitory concentration and minimal fungicidal concentration of cinnamaldehyde were 3 and 4 mmol l-1 on spore germination and colony development assays in vitro, respectively. Furthermore, the lesion diameter of potato tubers and tuber slices inoculated with F. sambucinum was reduced by 76·9 and 69% after treatment with 4 mmol l-1 cinnamaldehyde. Cytometric analyses revelled that cinnamaldehyde significantly affected the integrity of cell membrane firstly, then decreased mitochondrial membrane potential and induced the accumulation of intracellular reactive oxygen species. Meanwhile, high-performance liquid chromatography results indicated that 3 mmol l-1 cinnamaldehyde could reduce the ergosterol content by 67·94%. This effect was accompanied by a down-regulation of ERG11, ERG6 and ERG4 which were involved in ergosterol biosynthesis. CONCLUSION: Theses results suggest that cinnamaldehyde exerts strong antifungal activity against F. sambucinum, probably by affecting the ergosterol biosynthetic processes what leads to the disruption of cell membrane integrity. SIGNIFICANCE AND IMPACT OF THE STUDY: Cinnamaldehyde is a predominant constituent and key flavour compound of cinnamon essential oil. It has been used as a food additive and flavorant. It is expected to be a novel and safe fungicide for controlling dry rot in potato tubes.

[Experts consensus on the management of delirium in critically ill patients].

To establish the experts consensus on the management of delirium in critically ill patients. A special committee was set up by 15 experts from the Chinese Critical Hypothermia-Sedation Therapy Study Group. Each statement was assessed based on the GRADE (Grading of Recommendations Assessment, Development, and Evaluation) principle. Then the Delphi method was adopted by 36 experts to reassess all the statements. (1) Delirium is not only a mental change, but also a clinical syndrome with multiple pathophysiological changes. (2) Delirium is a form of disturbance of consciousness and a manifestation of abnormal brain function. (3) Pain is a common cause of delirium in critically ill patients. Analgesia can reduce the occurrence and development of delirium. (4) Anxiety or depression are important factors for delirium in critically ill patients. (5) The correlation between sedative and analgesic drugs and delirium is uncertain. (6) Pay attention to the relationship between delirium and withdrawal reactions. (7) Pay attention to the relationship between delirium and drug dependence/withdrawal reactions. (8) Sleep disruption can induce delirium. (9) We should be vigilant against potential risk factors for persistent or recurrent delirium. (10) Critically illness related delirium can affect the diagnosis and treatment of primary diseases, and can also be alleviated with the improvement of primary diseases. (11) Acute change of consciousness and attention deficit are necessary for delirium diagnosis. (12) The combined assessment of confusion assessment method for the intensive care unit and intensive care delirium screening checklist can improve the sensitivity of delirium, especially subclinical delirium. (13) Early identification and intervention of subclinical delirium can reduce its risk of clinical delirium. (14) Daily assessment is helpful for early detection of delirium. (15) Hopoactive delirium and mixed delirium are common and should be emphasized. (16) Delirium may be accompanied by changes in electroencephalogram. Bedside electroencephalogram monitoring should be used in the ICU if conditions warrant. (17) Pay attention to differential diagnosis of delirium and dementia/depression. (18) Pay attention to the role of rapid delirium screening method in delirium management. (19) Assessment of the severity of delirium is an essential part of the diagnosis of delirium. (20) The key to the management of delirium is etiological treatment. (21) Improving environmental factors and making patient comfort can help reduce delirium. (22) Early exercise can reduce the incidence of delirium and shorten the duration of delirium. (23) Communication with patients should be emphasized and strengthened. Family members participation can help reduce the incidence of delirium and promote the recovery of delirium. (24) Pay attention to the role of sleep management in the prevention and treatment of delirium. (25) Dexmedetomidine can shorten the duration of hyperactive delirium or prevent delirium. (26) When using antipsychotics to treat delirium, we should be alert to its effect on the heart rhythm. (27) Delirium management should pay attention to brain functional exercise. (28) Compared with non-critically illness related delirium, the relief of critically illness related delirium will not accomplished at one stroke. (29) Multiple management strategies such as ABCDEF, eCASH and ESCAPE are helpful to prevent and treat delirium and improve the prognosis of critically ill patients. (30) Shortening the duration of delirium can reduce the occurrence of long-term cognitive impairment. (31) Multidisciplinary cooperation and continuous quality improvement can improve delirium management. Consensus can promote delirium management in critically ill patients, optimize analgesia and sedation therapy, and even affect prognosis.

[Clinical utility study of 21-gene assay in 927 Chinese patients with early breast cancer].

Objective: To investigate the distribution patterns of 21-gene assay and its influencing factors in Chinese patients with early breast cancer. Methods: Nine hundred and twenty-seven early breast cancer patients were retrospectively recruited from January 2009 to December 2015 at Ruijin Hospital, Shanghai Jiaotong University School of Medicine. The 21-gene reverse transcriptase-polymerase chain reaction(RT-PCR) assay were conducted in paraffin-embedded tumor tissues to calculate the Recurrence Score(RS). Immunohistochemistry(IHC) assay was used to measure the expression levels of estrogen receptor(ER), progesterone receptor(PR) and Ki-67. Concordances of RT-PCR and IHC results were assessed. Correlations of RS and classical clinicopathological factors were evaluated, and logistic regression were applied to determine independent predictive factors for RS. Results: The median RS of 927 patients was 23(range: 0~90), and the proportions of patients categorized as having a low, intermediate, or high risk were 26.5%, 47.7% and 25.8%, respectively. The distribution of RS varied significantly according to different tumor grade, T stage, PR status, Ki-67 index and molecular subtypes(P<0.05 for all). Grade, PR status and Ki-67 index were independent predictive factors for RS. ER, PR status and Ki-67 index showed significantly correlation between RT-PCR and IHC assays, and the concordance rates for ER and PR status were 98.7% and 87.8%, respectively. Conclusions: RS significantly correlated with tumor grade, T stage, PR status, Ki-67 index and subtypes. Grade, PR status and Ki-67 index can independently predict RS. Remarkable concordances of ER, PR status and Ki-67 index are found between RT-PCR and IHC assays.

[Effect of 21-gene recurrence score on chemotherapy decisions for patients with estrogen receptor-positive, epidermal growth factor receptor 2-negative and lymph node-negative early stage-breast cancer].

Objective: To investigate the effect of 21-gene recurrence score on adjuvant chemotherapy decisions for patients with estrogen receptor (ER)-positive, epidermal growth factor receptor 2 (HER-2)-negative and lymph node (LN)-negative early stage-breast cancer. Methods: One hundred and forty-eight patients with ER+ , HER-2- and LN- early stage breast cancer were recruited in the Ruijin hospital, Shanghai Jiao Tong University School of Medicine. The 21-gene recurrence score (RS)assay was performed and systemic therapeutic decisions were made before and after knowing the RS results under multidisciplinary discussion. The effects of RS assay and the other influential factors on adjuvant chemotherapy decision were further analyzed. Results: After knowing the RS results, treatment decisions were changed in 26 out of 148 patients(17.6%). Among them, 9 out of 26 patients were not recommended for chemotherapy; 16 of 26 had treatment recommendation changed to chemotherapy, and chemotherapy regimen was changed in the last one patient. Multivariate analysis showed that RS, age and histological grade were independent factors of decision-making for adjuvant chemotherapy. Conclusion: Our results suggest that 21-gene recurrence score significantly influences decision making for adjuvant chemotherapy in patients with ER+ , HER-2- and LN- early stage breast cancer.

[Prognostic factors of postoperative delayed gastric emptying after pancreaticoduodenectomy: a predictive model].

Objective: To study the prognostic factors of delayed gastric emptying(DGE) after pancreaticoduodenectomy(PD) and construct a prognostic predictive model for clinical application. Methods: Clinic data of 401 consecutive patients who underwent PD between January 2012 and July 2016 in the First Affiliated Hospital of Harbin Medical University were retrospectively collected and analyzed. The patients were randomly selected to modeling group(n=299) and validation group(n=102) at a ratio of 3∶1. The data of modeling group were subjected to univariate and multivariate analysis for prognostic factors and to construct a prognostic predictive model of DGE after PD. The data of validation group were applied to test the prognostic predictive model. Results: DGE after PD occurred in 35 of 299 patients(11.7%) in the modeling group. The multivariate analysis of the modeling group showed that upper abdominal operation history(χ(2)=6.533, P=0.011), diabetes mellitus(χ(2)=17.872, P=0.000), preoperative hemoglobin <90 g/L(χ(2)=14.608, P=0.000) and pylorus-preserving pancreaticoduodenectomy(PPPD)(χ(2)=8.811, P=0.003) were associated with DGE after PD independently. A prognostic predictive model of DGE after PD was constructed based on these factors and successfully tested. The area under the receiver operating characteristic(ROC) curve was 0.761(95%CI: 0.666-0.856) of the modeling group and 0.750(95% CI: 0.577-0.923) of the validation group. Conclusions: Upper abdominal operation history, diabetes mellitus, preoperative hemoglobin<90 g/L and PPPD are associated with DGE after PD independently. The preoperative assessment of a patient's prognostic for DGE after PD is feasible. The model is a valid tool to take precautions against DGE after PD.

Aggravation of spinal cord compromise following new osteoporotic vertebral compression fracture prevented by teriparatide in patients with surgical contraindications.

Patients with spinal cord deficits following new unstable osteoporotic compression fracture and surgical contraindications were considered to receive conservative treatment. Teriparatide was better than alendronate at improving bone mineral density and bone turnover parameters, as well as preventing aggravation of spinal cord compromise. INTRODUCTION: This study compared the preventive effects of teriparatide and alendronate on aggravation of spinal cord compromise following new unstable osteoporotic vertebral compression fracture (OVCF) in patients with surgical contraindications. METHODS: This was a 12-month, randomized, open-label study of teriparatide versus alendronate in 49 patients with new unstable OVCF and surgical contraindications. Neurological function was evaluated using modified Japanese Orthopedic Association (mJOA) score (11-point scale, the maximum score of 11 implies normalcy). Visual analog scale (VAS) scores, kyphotic angles, anterior-border heights and diameters of the spinal canal of the fractured vertebrae, any incident of new OVCFs (onset of OVCF during follow-up), spine bone mineral density (BMD), and serum markers of bone resorption and bone formation were also examined at baseline and 1, 3, 6, and 12 months after initiation of the medication regimen. RESULTS: At 12 months, mean mJOA score had improved in the teriparatide group and decreased in the alendronate group. Mean concentrations of bone formation and bone resorption biomarkers, mean spine BMD, and mean anterior-border height and spinal canal diameter of the fractured vertebrae were significantly greater in the teriparatide group than in the alendronate group. Mean VAS score, mean kyphotic angle of the fractured vertebrae, and incidence of new OVCFs were significantly smaller in the teriparatide group than in the alendronate group. CONCLUSIONS: In patients with neurological deficits following new unstable OVCF and with surgical contraindications, teriparatide was better than alendronate at improving the BMD and the bone turnover parameters, as well as preventing aggravation of spinal cord compromise.

Molecular identification and genetic variation of varieties of Styphnolobium japonicum (Fabaceae) using SRAP markers.

Thirty-four Styphnolobium japonicum varieties were analyzed using sequence-related amplified polymorphism (SRAP) markers, to investigate genetic variation and test the effectiveness of SRAP markers in DNA fingerprint establishment. Twelve primer pairs were selected from 120 primer combinations for their reproducibility and high polymorphism. We found a total of 430 amplified fragments, of which 415 fragments were considered polymorphic with an average of 34.58 polymorphic fragments for each primer combination. The percentage of polymorphic fragments was 96.60%, and four primer pairs showed 100% polymorphism. Moreover, simple matched coefficients ranged between 0.68 and 0.89, with an average of 0.785, indicating that the genetic variation among varieties was relatively low. This could be because of the narrow genetic basis of the selected breeding material. Based on the similarity coefficient value of 0.76, the varieties were divided into four major groups. In addition, abundant and clear SRAP fingerprints were obtained and could be used to establish DNA fingerprints. In the DNA fingerprints, each variety had its unique pattern that could be easily distinguished from others. The results demonstrated that 34 varieties of S. japonicum had a relatively narrow genetic variation. Hence, a broadening of the genetic basis of breeding material is necessary. We conclude that establishment of DNA fingerprint is feasible by means of SRAP markers.

[Factors related to adjuvant ovarian function suppression in premenopausal breast cancer patients].

OBJECTIVE: To analyze the applied condition of ovary function suppression (OFS) before and after joint analysis of TEXT and SOFT trials and SOFT trial, and to identify the relevant factors of OFS usage. METHODS: The analysis was performed in premenopausal women with hormone receptor (HR) positive breast cancer receiving surgical treatment from Apr 2013 to Oct 2015 in Ruijin Hospital, Shanghai Jiaotong University School of Medicine. Adjuvant treatment strategy was made in the multidisciplinary team (MDT) meetings. We analyzed the applied condition of OFS before and after joint analysis, SOFT trial and its relevant factors. RESULTS: Among 454 patients, 114 (25.1%) patients received OFS. Before the results of joint analysis came out, all the patients (38/38) received OFS together with tamoxifen (TAM); after the results came out, clinicians began to put OFS with exemestane into practice, among 76 patients, 41(53.9%) patients received OFS with exemestane while 35 (46.1%) patients received OFS together with TAM. Before the results of SOFT trial came out, 71 out of 310 (22.9%) patients received OFS while 43 out of 144 (29.9%) patients received OFS after that. No significant difference was found between the proportion of patients receiving OFS before and after the results of SOFT trial came out (P=0.112). Age, histological grade, pN status, Ki-67 status, molecular subtype and acceptance of chemotherapy were correlated with OFS treatment (P<0.05). Age, tumor grade and pN were independent significant predictors of OFS usage. CONCLUSIONS: After the results of joint analysis came out, clinicians began to apply OFS with exemestane to premenopausal women with HR positive breast cancer. There is no significant difference between the proportion of patients receiving OFS before and after SOFT trial. Age, tumor grade and pN status are independent significant predictors of OFS treatment. Patients younger than 40, with histological grade Ⅱ or Ⅲ tumor and with pN1 or pN2 status are prone to receive OFS.

[Mutation screening of 433 families with Duchenne/Becker muscular dystrophy].

OBJECTIVE: Mutation analysis of unrelated families with Duchenne/Becker muscular dystrophy (DMD/BMD) was performed to investigate the characteristic of DMD gene mutation, especially the distribution pattern of point mutation of DMD gene in Chinese population. METHODS: A total of 433 unrelated DMD/BMD families were collected at the Center of Prenatal Diagnosis of the First Affiliated Hospital of Zhengzhou University from March 2010 to December 2014. The deletions or duplications in 79 exons of DMD gene were screened using multiplex ligation-dependent probe amplification (MLPA). Any single-exon deletion detected by MLPA was further validated by PCR amplification. In the 117 unrelated Chinese families in which large-scale deletions and duplications had been excluded by MLPA, the point mutation in 79 exons of DMD gene were tested in the propositus using next-generation sequencing (NGS), and further verified the point mutation using Sanger sequencing. RESULTS: In the 433 unrelated DMD/BMD families, 316 families with DMD deletions/duplications were identified by MLPA. Out of 57 single-exon deletions detected by MLPA, 3 were found as point mutations by PCR and Sanger sequencing, including 2 nonsense mutation (c.1729G>T [p.Glu577X], c. 3346A>T [p.Lys1116X]) and 1 frame-shift mutation (c.8605_8606delGT [p.Val2869ThrfsX25]). Direct sequencing with Ion PGM and Sanger sequencing in 117 families with negative results in MLPA detected 92 different point mutations in 96 families, including 46 novel mutations, 42 previously reported ones, and 4 possible polymorphisms (rs189143447, rs202008454, rs200213555, rs187617705). The 46 novel mutations consisted of 16 nonsense mutations (c.100A>T [p.Lys34X], c. 1201C>T [p.Gln401X], c. 1707C>A [p.Cys569X], c. 1831G>T [p.Glu611X], c. 1912C>T [p.Gln638X], c. 2213C>G [p.Ser738X], c. 3673_3673delA [p.Ile1225X], c. 3774C>A [p.Cys1258X], c. 4858G>T [p.Glu1620X], c. 5764A>T [p.Lys1922X], c. 6035T>G [p.Leu2012X], c. 6408G>A [p.Trp2136X], c. 7717C>T [p.Gln2573X], c. 7864G>T [p.Glu2622X], c. 8184_8185insT [p.Lys2729X], c. 8215C>T [p.Gln2739X]), 5 missense mutations (c.139G>A [p.Gly47Arg], c. 238G>C [p.Ala80Pro], c. 335G>T [p.Trp112Leu], c. 804A>C [p.Leu268Phe], c. 1149G>T [p.Glu383Asp]), 6 splice-site mutations (c.2293-3C>A, c. 2380+ 1G>T, c. 3277-1G>C, c.4519-7A>G, c. 5740-15G>T, c. 7661-1G>C), 16 small deletions (c.688_688delA [p.Met230CysfsX14], c.1760_1791del32 [p.Thr587IlefsX37], c. 2271_2271delA [p.Asp774ThrfsX22], c. 2281_2285delGAAAA [p.Glu761SerfsX10], c. 2527_2527delG [p.Glu843SerfsX3], c. 3405_3405delC [p.Asn1135LysfsX18], c. 4450_4450delC [p.His1484ThrfsX14], c. 4770_4770delA [p.Thr1590ThrfsX5], c. 4937_4937delA [p.Glu1646GlyfsX11], c. 5253_5256delATTA [p.Lys1751LysfsX2], c. 5654_5654delA [p.Gln1885ArgfsX6], c. 7441_7441delG [p.Glu2481AsnfsX13], c. 7860_7860delC [p.Ile2620IlefsX18], c. 8668-8668delG /c.8668+ 1-8668+ 1delG, c. 9009_9009delC [p.Thr3003ThrfsX18], c. 9021_9021delT [p.Ile3007IlefsX14]), and 3 small insertions (c.305_306insG [p.Gly102GlyfsX4], c. 3116_3117insA [p.His1039GlufsX11], c. 9197_9198insATCTC [p.Ser3066SerfsX25]). And 87.4% (83/95) of the pathologic point mutations disrupted the translational reading frame (46 nonsense mutations, 24 frame-shift mutations, and 13 splice-site mutations). CONCLUSIONS: Inexpensive and efficient genetic/prenatal diagnosis of DMD/BMD may be plausible by MLPA analysis, NGS, and Sanger sequencing. Most of the mutations identified in this study led to a predictable premature stop codon or splicing defects, resulting in defective function of dystrophin.

Analysis and application of ATP7B gene mutations in 35 patients with hepatolenticular degeneration.

We investigated the genetic mutations involved in Wilson's disease to improve prenatal genetic diagnosis and presymptomatic diagnosis. The polymerase chain reaction (PCR) was used to amplify the exons and exon-intron boundaries of the ATP7B gene in 35 Wilson's disease pedigrees. The PCR products were further analyzed by Sanger sequencing. Prenatal genetic diagnoses were performed by chorionic villus sampling after the genotypes of parents of the probands were identified. The overall mutation detection frequency was 92.9%. A total of 24 distinct mutations were detected, seven of which are novel: A1291T (c.3871G>A), c.2593_2594insGTCA, c.2790_2792delCAT, c.3661_3663delGGG, c.3700delG, c.4094_4097delCTGT, and IVS6+1G>A. Three mutations, R778L (c.2333G>T) (45.7%), A874V (c.2621C>T) (7.1%), and P992L (c.2975C>T) (7.1%) are relatively frequent. Two presymptomatic patients were detected through familial screening, and they began taking medicine after diagnosis. Of the subjects with Wilson's disease pedigrees who had received a prenatal genetic diagnosis, three fetuses were normal and one was a carrier. Twenty-four distinct mutations were identified, and our knowledge of the population genetics of Wilson's disease in China has therefore improved. For pedigrees with the Wilson's disease, genetic counseling, prenatal diagnosis, and presymptomatic diagnosis by Sanger sequencing and haplotype analysis are feasible.

Exogenous nitric oxide negatively regulates the S-nitrosylation p38 mitogen-activated protein kinase activation during cerebral ischaemia and reperfusion.

AIMS: A number of studies have suggested that nitric oxide (NO) plays an important role in the reactive phosphorylation of p38MAPKα (p38). However, whether S-nitrosylation of p38 is activated by NO and the details remain unclear. The aim of the present work was to assess the activation of p38, the S-nitrosylation site and the p38 signalling pathway in rat hippocampus and in HEK293 cell induced by exogenous NO. METHODS: Primary hippocampal cultures, HEK293 cells and rat model of cerebral ischaemia/reperfusion (brain ischaemia was induced by four-vessel occlusion procedure) were used in this study. Biotin-switch method and immunoblotting were performed to study the S-nitrosylation and phosphorylation of p38, and neuronal loss was observed by histology. RESULTS: Endogenous NO increased p38 phosphorylation and S-nitrosylation, and the activation of p38 was dependent on the S-nitrosylation of Cys-211, which was critical for the NO-mediated activation of p38. The exogenous NO donor sodium nitroprusside, S-nitrosoglutathione, 7-nitroindazole, the inhibitor of the neuronal nitric oxide synthase, inhibited the activation of p38 signal pathway induced by cerebral ischaemia/reperfusion and attenuated the damage in rat hippocampal neurones. Moreover, the N-methyl-D-aspartate receptor (NMDAR) is probably involved in the p38 activation process of S-nitrosylation and phosphorylation. CONCLUSION: Endogenous NO induces the S-nitrosylation and phosphorylation of p38 and mediates p38 signalling pathway by NMDAR, and as exogenous NO inhibits this process and is neuroprotective in rat cerebral ischaemia/reperfusion, it may make a contribution to stroke therapy.

Fire and flood management of coastal swamp enabled first rice paddy cultivation in east China.

The adoption of cereal cultivation was one of the most important cultural processes in history, marking the transition from hunting and gathering by Mesolithic foragers to the food-producing economy of Neolithic farmers. In the Lower Yangtze region of China, a centre of rice domestication, the timing and system of initial rice cultivation remain unclear. Here we report detailed evidence from Kuahuqiao that reveals the precise cultural and environmental context of rice cultivation at this earliest known Neolithic site in eastern China, 7,700 calibrated years before present (cal. yr bp). Pollen, algal, fungal spore and micro-charcoal data from sediments demonstrate that these Neolithic communities selected lowland swamps for their rice cultivation and settlement, using fire to clear alder-dominated wetland scrub and prepare the site for occupation, then to maintain wet grassland vegetation of paddy type. Regular flooding by slightly brackish water was probably controlled by 'bunding' to maintain crop yields. The site's exploitation ceased when it was overwhelmed by marine inundation 7,550 cal. yr bp. Our results establish that rice cultivation began in coastal wetlands of eastern China, an ecosystem vulnerable to coastal change but of high fertility and productivity, attractions maximized for about two centuries by sustained high levels of cultural management of the environment.

Stray light correction algorithm for multichannel hyperspectral spectrographs.

An algorithm is presented that corrects a multichannel fiber-coupled spectrograph for stray or scattered light within the system. The efficacy of the algorithm is evaluated based on a series of validation measurements of sources with different spectral distributions. This is the first application of a scattered-light correction algorithm to a multichannel hyperspectral spectrograph. The algorithm, based on characterization measurements using a tunable laser system, can be extended to correct for finite point-spread response in imaging systems.