ObjectiveThis study aimed to analyze the difference in setup error before and after correction of systematic error. To determine the most appropriate image-guided strategy during HT treatment， we use different scanning ranges and image-guidance frequencies in patients with nasopharyngeal carcinoma （NPC） treated with helical tomotherapy （HT）. MethodsFifteen patients with NPC who received HT treatment in Sun Yat-sen University Cancer Center from October 2019 to February 2020 were selected. Megavoltage computed tomography （MVCT） scanning was performed before each treatment. After five times of radiotherapy， system-error correction was performed to adjust the setup center. The setup errors before and after the correction of systematic errors， as well as the setup errors of different scanning ranges and different scanning frequencies， were collected for analysis and comparison. ResultsWhen comparing the setup errors before and after the correction of systematic error， the differences in setup errors in the left–right （LR）， superior–inferior （SI）， and anterior–posterior （AP） directions were statistically significant （P<0.05）.The different scanning ranges of "nasopharynx + neck" and "nasopharynx" were compared， and a statistically significant difference was found in yaw rotational errors （P<0.05）. In the comparison of daily and weekly scan frequency after system-error correction， a significant difference was found in AP direction （P<0.05）. ConclusionDuring radiotherapy for NPC， the systematic error can be corrected according to the first five setup errors， and then small-scale scanning was selected for image-guided radiotherapy every day.

AIM To investigate the effects of Chaihu Shugan Powder on the behaviors rat models of post-stroke depression(PSD)and the expression of 5-HT2A and BDNF in prefrontal cortex and striatum.METHODS The rats were randomly divided into the blank group,the sham operation group,the model group,Chaihu Shugan Powder group,the fluoxetine group and the combination drug group,with 9 rats in each group.The PSD rat models were established by the right middle cerebral artery occlusion(MCAO)by thread embolism method followed by further exposure to 4 weeks chronic and unpredictable mild stimulation.Subsequently,the rats started their corresponding regime of normal saline 5 mL/kg,Chaihu Shugan Powder 2 mg/kg,fluoxetine 12.5 mg/kg,and Chaihu Shugan Powder 2 mg/kg and fluoxetine 12.5 mg/kg(the combination group),respectively,and had their body mass measured and their behaviors(sugar water preference test,open field test and Morris water maze test)assessed simultaneously during the trial.After the experiment,the rats had their 5-HT2A and BDNF in prefrontal cortex and striatum detected of the mRNA expressions by RT-qPCR;of their expressions of the positive cells by immunohistochemical staining;and of their protein expressions by Westren blot.RESULTS Compared with the blank group,the model group displayed no improvement of depression-like behaviors,and decreased in mRNA expressions,positive cell expressions and protein expressions of 5-HT2A and BDNF(P<0.01).Compared with the model group,the groups intervened with Chaihu Shugan Powder,fluoxetine and combination drug showed improved depression-like behaviors,and increased mRNA expressions,positive cell expressions and protein expressions of 5-HT2A and BDNF as well(P<0.01).CONCLUSION Chaihu Shugan Powder can improve the depression-like behaviors in the PSD rat model,and its mechanism may be associated with the improvement in the expressions of 5-HT2A and BDNF in prefrontal cortex and striatum.

Hypertensive disorder of pregnancy (HDP) involves two major public health issues: mother-infant safety and prevention and controlling major chronic disease. HDP poses a serious threat to maternal and neonatal safety, and it is one of the leading causes of maternal and perinatal morbidity and mortality worldwide, as well as an important risk factor for long-term cardiovascular disease (CVD). In order to explore effective strategies to prevent and control the source of CVD and reduce its risk, we have established a cohort of HDPs in Shenzhen for the primordial prevention of CVD. The construction of the HDP cohort has already achieved preliminary progress till now. A total of 2 239 HDP women have been recruited in the HDP cohort. We have established a cohort data management platform and Biobank. The follow-up and assessment of postpartum cardiovascular metabolic risk in this cohort has also been launched. Our efforts will help explore the pathophysiological mechanism of HDP, especially the pathogenesis and precision phenotyping, prediction, and prevention of pre-eclampsia, which, therefore, may reduce the risk of adverse pregnancy outcomes, and provide a bridge to linking HDP and maternal-neonatal cardiovascular, metabolic risk to promote the cardiovascular health of mothers and their infants.

Objective:To develop a single-tube one-step multiplex nested real-time reverse transcription polymerase chain reaction (RT-PCR) assay for the simultaneous detection of 2019-nCoV, influenza A virus, influenza B virus and internal-control with human-derived gene.Methods:This study included 30 positive specimens for 2019-nCoV nucleic acid detection and 336 screening specimens collected from the arrivals at Guangzhou Baiyun Airport between February 2020 and February 2022. Sixty-four positive specimens of other respiratory pathogens were also collected from the arrivals at Guangzhou Baiyun Airport during the three-year period before the occurrence of COVID19 outbreak in 2020, and 7 positive viral strains of respiratory pathogens were provided by collaborative laboratories. In order to establish a set of multiplex nested real-time RT-PCR assay, a group of primers and probe combinations for a multiplex nested real-time RT-PCR was designed and screened according to a selection of nucleotide conserved regions of the ORF and N genes of 2019-nCoV and the M gene of influenza A and B viruses, while nested amplification primers and probe for the internal-control with human-derived gene were introduced. Then the prepared pseudovirus-positive quality control and sample discs were applied to evaluate the sensitivity and specificity. Clinical specimens were performed to validate the applicability of the method.Results:The results show that the established one-step multiplex nested real-time RT-PCR assay can specifically detect 2019-nCoV and influenza A and B viruses, with the limit-of-detection of about 125 copies/ml for 2019-nCoV and about 250 copies/ml for influenza A and B viruses. Totally 101 positive samples of various respiratory pathogens were detected, showing that the detection sensitivities of 2019-nCoV and influenza A and B viruses were 96.67%, 92.86% and 96.15%, respectively, with the specificity of 100%. No false-positive detection was found in the applied detection of more than 300 clinical samples.Conclusions:A one-step multiplex nested real-time RT-PCR assay for 2019-nCoV, influenza A and B viruses and human-derived gene internal-control was developed. The assay has good sensitivity and specificity and can be used for rapid screening of 2019-nCoV and influenza A and B viruses in high-volume samples.

Objective:To explore the effect of Bushen Huatan prescription on serum lipopolysaccharide （LPS） and Toll-like receptor 4 （TLR4）/ myeloid cell differentiation protein 88 （MyD88）/nuclear transcription factor-κB （NF-κB） signaling pathway in rats with ovariectomy-induced osteoporosis. Method:Sixty SPF 6-month-old female rats were randomly divided into sham operation group， model group， estradiol valerate group and Bushen Huatan prescription low， medium and high dose groups.One week after modeling by bilateral ovariectomy， 8 rats in each group were selected to receive intragastric administration.The estradiol valerate group was given 0.184 mg·kg^-1 by gavage， and Bushen Huatan prescription low， middle and high dose groups were given 4.7， 9.4 and 18.8 g·kg^-1 by gavage， sham operation group and model group were given 0.9% saline 4 mL by gavage respectively.After 12 weeks of intervention， the rats were sacrificed for detection.Serum LPS was detected by enzyme linked immunosorbent assay （ELISA）， while protein expressions of TLR4， MyD88 and phosphorylated （p）-NF-κB p65 in bone tissue were detected by Western blot， and the mRNA expressions of TLR4， MyD88， NF-κB p65， IL-1β， and IL-6 in bone tissue were detected by quantitative real-time polymerase chain reaction（PCR）. Result:Compared with sham operation group， the serum LPS level as well as protein expression of TLR4， MyD88， p-NF-κB p65 and mRNA expression of TLR4， MyD88， NF-κB p65， IL-1β， and IL-6 significantly increased in model group（P<0.05）.Compared with the model group， serum LPS level， protein expression of TLR4， MyD88， and p-NF-κB p65， mRNA levels of TLR4， MyD88， and NF-κB p65 in bone tissues as well as downstream inflammatory factors IL-1β， IL-6 mRNA expression decreased to different degrees in estradiol valerate group and Bushen Huatan prescription high dose group（P<0.05）. Conclusion:Bushen Huatan prescription can reduce serum LPS content， regulate mRNA and protein expression of TLR4， MyD88， NF-κB p65 and p-NF-κB p65 in TLR4/MyD88/NF-κB pathway， and down-regulate mRNA levels of IL-1β and IL-6 in bone tissues to improve bone microstructure and inhibit the development of postmenopausal osteoporosis （PMOP）.

Objective To investigate the genetic polymorphisms of Plasmodium falciparum multidrug resistance protein 1 (PfMDR1), chloroquine resistance transporter (PfCRT) and Kelch 13 (PfK13) genes in Bioko Island, Equatorial Guinea, so as to provide insights into the development of the malaria control strategy in local areas. Methods A total of 85 peripheral blood samples were collected from patients with Plasmodium falciparum infections in Bioko Island, Equatorial Guinea in 2018 and 2019, and genomic DNA was extracted. The PfMDR1, PfCRT and PfK13 genes were amplified using a nested PCR assay. The amplification products were sequenced, and the gene sequences were aligned. Results There were no mutations associated with artemisinin resistance in PfK13 gene in Bioko Island, Equatorial Guinea, while drug-resistant mutations were detected in PfMDR1 and PfCRT genes, and the proportions of PfMDR1_N86Y, PfMDR1_Y184F and PfCRT_K76T mutations were 35.29% (30/85), 72.94% (62/85) and 24.71% (21/85), respectively. Conclusion There are mutations in PfMDR1, PfCRT and PfK13 genes in P. falciparum isolates from Bioko Island, Equatorial Guinea.

[Objective]To explore the different scanning frequency for the MVCT scanning in the tomotherapy , and seek the best strategy of the scanning.[Methods]Patients were treated by tomotherapy in SYSUCC from January 1, 2015 to May 15, 2016, includ?ing 88 head&neck patients, 63 chest&abdomen patients and 19 pelvis patients. All patients have had a daily MVCT scanning for po?sition verification before the treatment. The data retrieved from"the first five times of the very first week"and"the every first time of each week", were compared with"the whole course"data, respectively.[Results]There are no significant difference between"the whole course"and"the first five times of the very first week"or"the every first time of each week"for pelvis patients and chest and abdomen patients. There are significant differences between Z axis of"the whole course"and"the first five times of the very first week"only for head&neck patients.[Conclusion]It is a good scanning choice of"the first five times of the very first week"and"the every first time of each week"for the those patients with thoracic&abdominal or pelvic tumors to reduce the extra radiation dose from the MVCT. There are significant differences between Z axis of"the whole course"and"the first five times of the very first week"in the head&neck patients. Therefore,"the first five times of the very first week"cannot be used as a solid position information for daily verification. And"the every first time of each week"may be a good compromise solution.

Objectives To explore the clinical significance of preoperative abdominal multislice spiral CT angiography (MSCTA) in radical resection of gastric cancer. Methods One hundred and three patients with gastric cancer were divided into two groups according to their desires. Group Ⅰ included 57 patients who underwent preoperative MSCTA and group Ⅱ included 46 patients who underwent surgery without preoperative MSCTA. All these patients were operated by the same surgical team. Results Six patients (10.5%) with abnormal gastric artery in group Ⅰ were discovered. The diagnostic concordance rate between MSCTA and intraoperative findings was 100% in group Ⅰ in the locations and alignments of main perigastric vessels and their relationship with cancer lesions.Operative time in group Ⅰ was shorter than that in group Ⅱ [(206±23)min vs. (257±32)min, P=0.044]. Operative time [(190±50) min] of patients with abnormal gastric artery of group Ⅰ was shorter than that [(255±62) min] of patients with abnormal gastric artery discovered during operation of group Ⅱ (P=0.048). However there were no differences in blood loss, extent of lymph node dissection,complication rate, length of hospital stay, and hospitalization cost between the two groups (P＞0.05).Conclusion Preoperative MSCTA is beneficial to the evaluation of vascular structure of the cancer and the adjacent tissues, which may reduce postoperative complications.

Objectives To explore the clinical significance of preoperative abdominal multislice spiral CT angiography (MSCTA) in radical resection of gastric cancer. Methods One hundred and three patients with gastric cancer were divided into two groups according to their desires. Group Ⅰ included 57 patients who underwent preoperative MSCTA and group Ⅱ included 46 patients who underwent surgery without preoperative MSCTA. All these patients were operated by the same surgical team. Results Six patients (10.5%) with abnormal gastric artery in group Ⅰ were discovered. The diagnostic concordance rate between MSCTA and intraoperative findings was 100% in group Ⅰ in the locations and alignments of main perigastric vessels and their relationship with cancer lesions.Operative time in group Ⅰ was shorter than that in group Ⅱ [(206±23)min vs. (257±32)min, P=0.044]. Operative time [(190±50) min] of patients with abnormal gastric artery of group Ⅰ was shorter than that [(255±62) min] of patients with abnormal gastric artery discovered during operation of group Ⅱ (P=0.048). However there were no differences in blood loss, extent of lymph node dissection,complication rate, length of hospital stay, and hospitalization cost between the two groups (P＞0.05).Conclusion Preoperative MSCTA is beneficial to the evaluation of vascular structure of the cancer and the adjacent tissues, which may reduce postoperative complications.

Armillariella tabescens EJLY2098 was capable of secreting p-mannanase by konjac inducement. A 34 orthogonal design was applied to determine the optimum medium of inducing mannanase by Armillariella tabescens EJLY2098. The results suggested that Armillariella tabescens EJLY2098 secreted the high-activity enzyme in the optimum medium, which was composed of 2% konjac, 1% peptone, 25% potato juice,0.3% KH2PO4,15% MgSO4?7H2O, 0.01% VitB1. Purified by DEAE-anion exchange chromatography, two eluting peaks (P1 and P2) with the p-mannanase activity were obtained, and one of them (named?-mannanase P2) was a single band by the SDS-PAGE, and the molecular weight of?-mannanase P2 was 78. 9kDa. The isoelectric point of?-mannanase P2 was estimated to be 4.0-4. 1. The optimum activity for the enzyme was found at 60℃and pH4. 0 - 6. 0, and the enzyme was stable between pH4. 5 - 6. 0. The activity of?-mannanase P2 were enhanced by Na+ and Ba2+ . This?-mannanase can be used in feed industy. a new fungi secreting?-mannanase was obtained, providing an important base for cloning mannanase gene and constructing recombin microbe expressing?-mannanase .