Biosensors have become powerful tools for real-time monitoring of specific small molecules and precise control of gene expression in biological systems. High-throughput sensors for 1, 4-butanediamine biosynthesis can greatly improve the screening efficiency of high-yielding 1, 4-butanediamine strains. However, the strategies for adapting the characteristics of biosensors are still rarely studied, which limits the applicability of 1, 4-butanediamine biosensors. In this paper, we propose the development of a 1, 4-butanediamine biosensor based on the transcriptional regulator PuuR, whose homologous operator puuO is installed in the constitutive promoter P_gapA of Escherichia coli to control the expression of the downstream superfolder green fluorescent protein (sfGFP) as the reporter protein. Finally, the biosensor showed a stable linear relationship between the GFP/OD₆₀₀ value and the concentration of 1, 4-butanediamine when the concentration of 1, 4-butanediamine was 0-50 mmol/L. The promoters with different strengths in the E. coli genome were used to modify the 1, 4-butanediamine biosensor, and the functional properties of the PuuR-based 1, 4-butanediamine biosensor were explored and improved, which laid the groundwork for high-throughput screening of engineered strains highly producing 1, 4-butanediamine.
Biosensing Techniques/methods* ; Escherichia coli/metabolism* ; Promoter Regions, Genetic/genetics* ; Green Fluorescent Proteins/metabolism* ; Transcription Factors/genetics* ; Escherichia coli Proteins/genetics* ; Diamines/metabolism* ; Gene Expression Regulation, Bacterial

The correspondence between the five zang organs and the five phases exhibited variations in the previous and the current literature， which is related to the understanding and interpretation of the historical construction of the basic theories of traditional Chinese medicine， and is an important issue that has been debated for a long time in the history of academia but has not been resolved. Through combing and analyzing the relevant literature handed down during the pre-Qin and Han dynasties， which includes Master LYU's Spring and Autumn Annals （《吕氏春秋》）， The Book of Rites： Monthly Ordinances （《月令》）， Huainanzi （《淮南子》）， Records of the Grand Historian： Biography of BIAN Que and CANG Gong （《史记·扁鹊仓公列传》）， The Inner Canon of Yellow Emperor （《黄帝内经》）， as well as excavated literature like the Warring States bamboo slips Tang Zai Chi Men （《汤在啻门》） collected by Tsinghua University， Taichan Shu （《胎产书》） unearthed from the Mawangdui Han Dynasty Tombs， Tianhui Medical Manuscripts （《天回医简》） unearthed from the Han Dynasty Tombs of Lao Guan Mountain， Chengdu， this paper finds that the current five phases are newly proposed by previous medical practitioners on the basis of the objective understanding of the physiology and pathology of the five zang organs， which is a unique contribution of traditional Chinese medicine to the traditional Chinese culture. The lacquered acupuncture figurine unearthed from the Han Dynasty Tombs of Lao Guan Mountain， Tianhui Town， Chengdu， Sichuan Province， is engraved on its back with the inscriptions of "heart， lungs， liver， stomach， and kidneys"， which refer to the back-Shu points of the five zang organs， and the order follows the sequence of “fire - metal - wood - earth - water”， which is the order of the five phases being restricted. It is believed that this is a reflection of the idea of “heart is the head of the five organs” in the early medical scriptures， which can be used as a basis for judging the chronological order of the relevant chapters of the modern version of The Inner Canon of Yellow Emperor. This order is also seen in the Tianhui Medical Manuscripts unearthed from the same tombs as the acupuncture figurine and is preserved in many chapters of the modern version of The Inner Canon of Yellow Emperor.

Objective:This study aimed to explore the clinical value of myostatin(MST) and follistatin(FST) as biological biomarkers in evaluating sarcopenia in elderly women.Methods:This was a retrospective cross-sectional study that enrolled 350 females aged 20-89 years who underwent physical examinations in Shanghai Huadong Hospital in 2021. Demographic characteristics, muscle mass, fat mass, bone mineral density, hand grip strength, gait speed, and serum indices of MST and FST were collected.Results:The serum levels of MST did not change significantly with age. However, the serum levels of FST increased with age. In women aged≥60 years, MST was positively correlated with total lean mass and appendicular skeletal muscle index(ASMI; r=0.236, P=0.041; r=0.289, P=0.014), while FST was negatively correlated with ASMI( r=-0.265, P=0.030). In multivariate stepwise regression analysis, after adjusting for age, body mass index, hip bone mineral density, and total fat mass, only FST was independently correlated with ASMI( β=-0.238, P=0.006), while MST was not correlated with ASMI. The receiver operating characteristic curve was plotted using muscle mass reduction as the state variable and serum FST level as the test variable. The area under the curve was 0.753. And when the FST cutoff value was 17.49 ng/mL, the maximum Jordan index was 0.46, with a sensitivity of 77.3% and a specificity of 68.7%. Women aged ≥60 years were divided into three groups based on serum FST levels. Compared to the upper third of the serum FST level group, the low third of the FST level group had a significantly reduced risk of suffering from sarcopenia( OR=0.098, P =0.036). Conclusions:Serum FST lever has a better correlation with muscle mass among elderly women, making it a promising biomarker for evaluating muscle mass.

Objective:To observe any effects of contralateral repeated transcranial magnetic stimulation (rTMS) of the swallowing motor cortex on the swallowing and brainstem auditory evoked potentials (BAEPs) of stroke survivors with dysphagia.Methods:A total of 83 stroke survivors with dysphagia were randomly divided into an ipsilesional stimulation group ( n=22), a contralesional stimulation group ( n=21), a bilateral stimulation group ( n=20), and a control group ( n=20). In addition to their conventional dysphagia training, those in the three stimulation groups received 3Hz rTMS while the control group was given fake stimulation. The treatment was administered daily for 20 minutes, 6 days a week, for 5 consecutive weeks. Before and after the treatment, swallowing function was assessed videofluoroscopically and using the Dysphagia Outcome and Severity Scale (DOSS). The oral and pharyngeal stages of swallowing were evaluated using the videofluoroscopic dysphagia scale (VDS). Brain stem conduction was assessed using BAEPs. Results:After treatment the average DOSS scores of all 4 groups were significantly better than before the treatment. The average DOSS scores of the contralesional and bilateral sti-mulation groups were then significantly better than those of the other two groups. The sub-item and total VDS scores of all 4 groups had decreased significantly, but the average score of the bilateral stimulation group was significantly lower than the control group′s average. Ipsilesional stimulation significantly improved the VDS sub-item scores for the triggering of pharyngeal swallowing, laryngeal elevation, and pharyngeal transit time compared with the control group. In the contralesional stimulation group the average total score and the VDS sub-item scores for apraxia, premature bolus loss, oral transit times, the triggering of pharyngeal swallowing, vallecular residue, laryngeal elevation, coating on the pharyngeal wall, and pharyngeal transit time were significantly lower than those of the control group, on average. After the treatment the latencies of BAEP waves I, III and V and the I-III, III-V and I-V interpeak intervals had decreased significantly in all four groups, but the average latencies and intervals of the bilateral and contralesional groups were significantly shorter than those of the control group. The latencies and intervals of the bilateral stimulation group were then significantly shorter than those in the ipsilesional stimulation group on average. The average latency of wave V in the bilateral stimulation group (6.53±0.73ms) was significantly shorter than that in the contralesional stimulation group after the treatment.Conclusion:Bilateral rTMS over the swallowing motor cortex combined with conventional dysphagia training can significantly improve the swallowing of dysphagic stroke survivors.

Background: Crawling-type gastric adenocarcinoma is a rare subtype of gastric cancer, however, the understanding on this special entity of gastric cancer is still lacking. Aims: To investigate the clinical, endoscopic and pathological characteristics of crawling-type early gastric adenocarcinoma. Methods: Patients diagnosed as crawling-type early gastric adenocarcinoma in Songjiang Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from January 2016 to December 2021 were recruited consecutively in a retrospective study. The clinical data were reviewed, the pathological specimens were collected for immunohistochemical staining, and a telephone follow-up was conducted for prognosis analysis. Results: Fourteen patients with crawling-type early gastric adenocarcinoma and fulfilling the inclusion criteria were enrolled in the study, of them, 9 were male, 5 were female, and the mean age was 65.9 years old. No family history of gastric cancer was reported. The clinical manifestations were not specific. All patients were positive for Helicobacter pylori (Hp) infection. Tumors were more likely to occur in the middle and lower thirds of the stomach, with marked atrophic background mucosa. Macroscopically, 11 lesions were superficial-depressed (0-IIc) and 3 were superficial-flat type (0-IIb+ IIc). The color of the lesions was red. Lesions with indistinct border were observed endoscopically in 10 cases. Complete resection was achieved in all 14 patients after endoscopic submucosal dissection n=10 or surgical treatment n=4. Three submucosal and 11 intramucosal infiltration were observed pathologically. Immunohistochemical results of gastric (MUC5AC and MUC6) and intestinal (MUC2, CD10 and CDX-2) markers showed that most of the patients were mixed immunophenotype; the Ki-67 index ranged mostly between 30% and 70%. In a mean follow-up time of 38 months, all 14 patients were survived. Two patients with heterochronous early gastric cancer were found by follow-up endoscopy. Conclusions: When a superficial-depressed or superficial-flat type tumor with reddish color change and atrophic background mucosa is observed endoscopically in an Hp-positive patient, the possibility of crawling-type early gastric adenocarcinoma should be considered. Adequate preoperative evaluation should be carried out to judge the extent and depth of tumor invasion, which may guide the decision of treatment strategy. Postoperative endoscopic surveillance is recommended.

AIM: To investigate the relationship between human cholesteryl ester transfer protein CETP gene polymorphism and postoperative neurocognitive disorders (PND). METHODS: A total of 124 elderly patients over 65 years of age who underwent elective non-cardiac surgery were enrolled in the study while 25 healthy volunteers matching age and sex were recruited as the control group. Neuropsychological tests were performed 1 day before surgery, 7 days, and 3 months after surgery. PND was determined using the Z value method. The venous blood sample of the surgical patient was taken before the operation, followed by direct gene sequencing. Statistical methods were used to calculate the correlation between CETP gene polymorphism (rs5882) and PND. RESULTS: The incidence of PND was 29.3% and 18.2% at 7 days and 3 months after operation respectively. The A allele frequency of PND patients was significantly higher than that of non-PND patients 7 days and 3 months after surgery (65.52% vs. 41.43%, 34.48% vs. 58.57%, P=0.001), while the G allele frequency in PND group lower than that of non-PND (58.33% vs. 37.86%, 41.67% vs. 62.14%, P=0.004).AA genotype in PND patients was 34.48%, 38.89% at 7 days and 3 months after surgery respectively, significantly higher than 14.29%, 16.05% of non-PND (P=0.023, P=0.029). CONCLUSION: CETP rs5882 polymorphism is associated with PND and AA genotype may be a predisposing factor for postoperative PND in Chinese Han elderly patient.

Objective:To observe the effect of observing good swallowing on the swallowing action of stroke survivors with dysphagia.Methods:Eighteen stroke survivors with dysphagia were randomly divided into a treatment group ( n=9) and a control group ( n=9). In addition to routine swallowing rehabilitation therapy, the treatment group was asked to simulate swallowing after watching a video of normal people′s swallowing action. They did so 5 times a week for 10 minutes, while the control group just watched landscape videos at the same time. The treatment lasted 8 weeks. Before and after the treatment, both groups were assessed using the eating assessment tool (EAT-10), the functional oral intake scale (FOIS) and the penetration and aspiration scale (PAS). Functional magnetic resonance imaging (fMRI) was also used to observe their swallowing action. Results:There was no significant difference between the two groups in any of the measurements before the treatment. After the 8 weeks of treatment the average EAT-10, FOIS and PAS scores of the treatment group were all significantly better than before the treatment and better than the control group′s averages at the time. fMRI showed significantly more areas activated in the precuneus, parietal lobe, posterior central gyrus, BA7, BA5, frontal lobe and paracentral lobule in the treatment group compared with before the intervention and also more than in the control group.Conclusions:Observing proper swallowing action can improve dysphagia and activation of the swallowing-related brain areas of stroke survivors.

Objective:To observe the effect of applying repetitive transcranial magnetic stimulation (rTMS) on swallowing function and auditory evoked potentials in the brain-stems of stroke survivors with dysphagia.Methods:Sixty stroke survivors with dysphagia were randomly assigned to an observation group and a control group, each of 30. Both groups were given 30 minutes of routine swallowing training 6 times a week for 5 weeks, while the observation group was additionally provided with two-second bursts of rTMS on the left and right brain hemispheres alternately for 20 minutes daily. The frequency was 3.0Hz and the strength was 80% of the resting motor threshold (RMT), with an interval of 10 seconds between bursts. Before and after the treatment, both groups′ swallowing was assessed using the functional communication measure (FCM) and the modified Mann swallowing capacity evaluation scale (MMASA). Brainstem auditory evoked potentials (BAEP) were recorded to detect any changes in the peak latency (PL) and inter-peak latency (IPL). The clinical efficacy was also observed.Results:After the treatment, the average FCM results of both groups had improved significantly, but the observation group had improved significantly more. The average MMASA total scores and the average scores on all the sub-items had improved significantly more in the observation group except for the expressive speech disorder and anarthria scores. After the treatment, improvement was observed in respiration, anarthria, range of motion of the tongue muscles, strength of the tongue muscles, gag reflex and soft palate movement compared with before the intervention. Peak and inter-peak latencies had also improved significantly more in the observation than in the control group, on average.Conclusions:rTMS combined with conventional swallowing training can significantly improve the swallowing of dysphagic stroke survivors, and shorten the PL of the BAEP so as to better regulate deglutition.

Objective:To investigate the clinicopathological characteristics of gastric adenocarcinoma of the fundic gland(GA-FG).Methods:A total of 6 patients with GA-FG admitted from January 2014 to May 2019 were identified at Shanghai Songjiang District Central Hospital.Analyzed endoscopic findings, pathological characteristics and immunohistochemical staining of the lesions and reviewed relevant literatures.Results:Among the 6 patients, 3 were male, 3 were female, and the mean age was 66 years. The most common clinical presentation was epigastric discomfort. Gastroscopic examination revealed an elevated or flat lesion in the body of the stomach, with a diameter of 0.4 to 1.0 cm. Histopathologically, they mainly composed of main cells, but there were also scattered parietal cells, arranged in an irregular tubular or cord-like structure, and formed anastomosis branches. The tumor cells were immunohistochemically positive for MUC6 and pepsinogen 1.Conclusions:GA-FG is a rare variant of well-differentiated adenocarcinoma. It is important to understand its clinical features and histological morphology for more accurate diagnosis.

Objective To analyze the time of removing setons by observing 30 cases of the infliximab (IFX)combined drainage setons therapy in the treatment of perianal fistulising Crohn's disease.Methods Thirty patients of perianal fistulising Crohn's disease of Department of Colorectal Surgery,Jiangsu Prorince Hospital of TCM in Aug.2015 to Jul.2017 were treated with drainage setons followed by intravenous IFX injections according to the standard,and setons were removed after the third injection of IFX.The method of repeated measurement were adopted to record.Clinical symptoms,perianal manifestations,laboratory indexes,healing rate and general linear model and nonparametric test of SPSS 23.0 software were used for statistical analysis.Results The differences of WBC,CRP,ESR,HB,PLT,BMI,CDAI and PDAI between post treatment and pretreatment were statistically significant (P ＜ 0.05).WBC,CRP and ESR will rerise at some time after removing the rubber band (sixth week).The closure rate of fistular at the 14th week was 53.3％,and after the follow-up in the ward at 22th week and 30th week,the clinical efficacy of fistular was not statistically significant (P ＞0.05) at the 14th week,22th week and 30th week.Conclusion IFX combined drainage setons which was removed when the induction therapy finished can induce Crohn's disease remission,improve laboratory index and promote fistula closure.the clinical efficacy in short time was stable.