To construct stress-responsive promoters, we mined the transcriptome data of the industrial strain A223 under stress. The transcription factor CIN5 showed significantly increased expression under stress but exhibited limited resistance. Further analysis of CIN5-interacting genes revealed that the binding motif "TTACGTAATC" (named CIN5BS) of CIN5 displayed transcription-enhancing activity. Four artificially enhanced promoters TCIN5B(3-6) were created by insertion of CIN5BS as a cis-element into different sites of the promoter TEF1, achieving 15.25-fold transcriptional enhancement. Five cis-elements (CIN5B4-1-CIN5B4-5) were designed through G+C content optimization, generating five stronger artificially enhanced promoters (TCIN5B4-1-TCIN5B4-5). For example, TCIN5B4-1 demonstrated 4.71 times higher transcriptional activity than the control at 37 ℃. This study established a technical framework of transcription factor mining-cis-element design-promoter reconstruction, providing a reference strategy for yeast cell factories to stably produce natural compounds under high-temperature stress conditions.
Promoter Regions, Genetic/genetics* ; Transcription Factors/genetics* ; Saccharomyces cerevisiae/metabolism* ; Stress, Physiological/genetics* ; Saccharomyces cerevisiae Proteins/genetics*

With the increasing proportion of human papilloma virus(HPV)infection in the pathogenic factors of oro-pharyngeal cancer,a series of changes have occurred in the surgical treatment.While the treatment mode has been im-proved,there are still many problems,including the inconsistency between diagnosis and treatment modes,the lack of popularization of reconstruction technology,the imperfect post-treatment rehabilitation system,and the lack of effective preventive measures.Especially in terms of treatment mode for early oropharyngeal cancer,there is no unified conclu-sion whether it is surgery alone or radiotherapy alone,and whether robotic minimally invasive surgery has better func-tional protection than radiotherapy.For advanced oropharyngeal cancer,there is greater controversy over the treatment mode.It is still unclear whether to adopt a non-surgical treatment mode of synchronous chemoradiotherapy or induction chemotherapy combined with synchronous chemoradiotherapy,or a treatment mode of surgery combined with postopera-tive chemoradiotherapy.In order to standardize the surgical treatment of oropharyngeal cancer in China and clarify the indications for surgical treatment of oropharyngeal cancer,this expert consensus,based on the characteristics and treat-ment status of oropharyngeal cancer in China and combined with the international latest theories and practices,forms consensus opinions in multiple aspects of preoperative evaluation,surgical indication determination,primary tumor re-section,neck lymph node dissection,postoperative defect repair,postoperative complication management prognosis and follow-up of oropharyngeal cancer patients.The key points include:① Before the treatment of oropharyngeal cancer,the expression of P16 protein should be detected to clarify HPV status;② Perform enhanced magnetic resonance imaging of the maxillofacial region before surgery to evaluate the invasion of oropharyngeal cancer and guide precise surgical resec-tion of oropharyngeal cancer.Evaluating mouth opening and airway status is crucial for surgical approach decisions and postoperative risk prediction;③ For oropharyngeal cancer patients who have to undergo major surgery and cannot eat for one to two months,it is recommended to undergo percutaneous endoscopic gastrostomy before surgery to effectively improve their nutritional intake during treatment;④ Early-stage oropharyngeal cancer patients may opt for either sur-gery alone or radiation therapy alone.For intermediate and advanced stages,HPV-related oropharyngeal cancer general-ly prioritizes radiation therapy,with concurrent chemotherapy considered based on tumor staging.Surgical treatment is recommended as the first choice for HPV unrelated oropharyngeal squamous cell carcinoma(including primary and re-current)and recurrent HPV related oropharyngeal squamous cell carcinoma after radiotherapy and chemotherapy;⑤ For primary exogenous T1-2 oropharyngeal cancer,direct surgery through the oral approach or da Vinci robotic sur-gery is preferred.For T3-4 patients with advanced oropharyngeal cancer,it is recommended to use temporary mandibu-lectomy approach and lateral pharyngotomy approach for surgery as appropriate;⑥ For cT1-2N0 oropharyngeal cancer patients with tumor invasion depth＞3 mm and cT3-4N0 HPV unrelated oropharyngeal cancer patients,selective neck dissection of levels ⅠB to Ⅳ is recommended.For cN+HPV unrelated oropharyngeal cancer patients,therapeutic neck dissection in regions Ⅰ-Ⅴ is advised;⑦ If PET-CT scan at 12 or more weeks after completion of radiation shows intense FDG uptake in any node,or imaging suggests continuous enlargement of lymph nodes,the patient should undergo neck dissection;⑧ For patients with suspected extracapsular invasion preoperatively,lymph node dissection should include removal of surrounding muscle and adipose connective tissue;⑨ The reconstruction of oropharyngeal cancer defects should follow the principle of reconstruction steps,with priority given to adjacent flaps,followed by distal pedicled flaps,and finally free flaps.The anterolateral thigh flap with abundant tissue can be used as the preferred flap for large-scale postoperative defects.

Objective:To explore the genetic etiology of a child suspected for peroneal muscular atrophy.Methods:The child and his parents were analyzed by using next generation sequencing.Results:The child was found to harbor compound heterozygous variants of c. 52G>T (p.Glu18X) and c. 1390C>T (p.Arg464X) of the PRX gene, which were inherited from his father and mother, respectively. Among these, the c. 52G>T variant was previously unreported. Based on the standards and guidelines of the American College of Medical Genetics and Genomics, both variants were predicted to be pathogenic (PVS1+ PM2+ PM3, PVS1+ PM3-Strong+ PM2+ BS2). Conclusion:The compound heterozygous variants of the PRX gene probably underlay the Charcot-Marie-Tooth disease type 4F in this child. Above finding has enriched the mutational spectrum of the PRX gene.

Objective To establish an HPLC method for simultaneous assay of macrocyclic polyphenols from Penthorum chinense Pursh, pinocembrin-7-O-[4'', 6''-(S)-hexahydroxydiphenoyl]-β-D-glucose (PHG), pinocembrin-7-O-[3''-O-galloyl-4'', 6''-(S)-hexahydroxydiphenoyl]-β-D-glucose (PGHG) and pinocembrin dihydrochalcone-7-O-[3''-O-galloyl-4'', 6''-(S)-hexahydroxydiphenoyl]-β-D-glucoside or thonningianin A (THA), and optimize the extraction process. Methods The total extraction rate of PHG, PGHG, THA was used as an investigated index to analyze the extracts from Penthorum chinense Pursh. Orthogonal design was applied to evaluate solvent amount, extraction time, solvent concentration and extraction times as the influencing factors for the optimal extraction process of macrocyclic polyphenols from Penthorum chinense Pursh. Results When this content assay method was adopted, there were good linear relationships for PHG, PGHG, THA in the linear range. The recoveries were between 100.90% to 102.04% with the RSDs below 1.5%. The optimal extraction process was involved in cutting Penthorum chinense Pursh into 3-5 cm, adding 10 times 80% ethanol aqueous solution by volume and refluxing 2 hours twice. The extraction rate of macrocyclic polyphenols was above 90% with this process. Conclusion This assay method is accurate, stable, and repeatable. The optimized extraction process is stable and feasible for further development and utilization.

With the rapid development of information technology, artificial intelligence technology (AI) and how to use it have become the focus of current researches. The application of AI in the field of TCM has shown its uniqueness. The combination of artificial intelligence technology and traditional Chinese medicine provides a new direction and idea for the development of TCM. This paper analyzes the strengths, weaknesses, opportunities and threats (SWOT) of TCM development against the background of AI technology in China by SWOT analysis method. Based on these analyses, this paper puts forward some counter measures such as protecting personal information, avoiding negative effects and medical regulations etc. In order to facilitate and guarantee the development of TCM, we should take advantage of the AI, and avoid its disadvantages.

Objective: To compare the survival rates difference between diabetic kidney disease (DKD) and non-DKD maintenance hemodialysis patients. Methods: The eligible patients who started hemodialysis treatment in Dalian Municipal Central Hospital from January 1, 2010 to December 31, 2016 were enrolled. The endpoint was all-cause death. Patients were divided into two groups according to the primary disease: DKD group and non-DKD group. Survival between two groups was compared by Kaplan-Meier plots and log-rank test. Survival was timed from the start of dialysis until the date of death and was censored for the date of end of the study period (December 31, 2016). SPSS 13.0 software was used for statistical analysis. Univariate COX regression analysis was used for risk assessment. Independent analysis was performed by multivariate COX regression. P < 0.05 indicated that the difference was statistically significant. Results: A total of 769 patients were enrolled, including 305 patients with DKD (39.7%) and 464 patients with non-DKD (60.3%). There were 465 males, accounting for 60.5%, and 304 females, accounting for 39.5%. The mean age of starting dialysis was (56.2 ± 14.9) years. The median follow-up time was 21 months. One hundred and seventy patients died due to all causes, accounting for 21.7%. The 1-, 2-, 3-, 4-, 5-, 6- and 7-year survival rates in the diabetic kidney disease group were 94%, 77%, 68%, 56%, 44%, 31% and 26%. The 1-, 2-, 3-, 4-, 5-, 6- and 7-year survival rates in the non-diabetic kidney disease group were 94%, 87%, 81%, 77%, 69%, 65% and 60%. The survival rate of DKD group was significantly lower than that of non-DKD group (χ²=23.656, P < 0.01). Multivariate Cox regression analysis showed that age of onset of dialysis, primary disease, low density lipoprotein, serum potassium, ejection fraction (EF), coronary heart disease and stroke were independent risk factors of mortality (P < 0.05). Conclusions The survival rate of patients with diabetic kidney disease is significantly lower than that of patients with non-diabetic kidney disease in the maintenance hemodialysis patients in our center. Age, primary disease, low density lipoprotein, EF, coronary heart disease, and stroke are independent predictors of death.

Objective To investigate the length of preserved ileocecum in surgical treatment of slow transit constipation ( STC) by sub-total colectomy with antiperistaltic cecorectal anastomosis .Methods A total of 82 patients with STC were divided into two groups according to the random number table method ,with 41 cases in each group ,all the patients of the two groups underwent subtotal colectomy ,intraopera-tive ileocecum was preserved length of group A was 10~15 cm,group B was 2~3 cm.The operation time,intraoperative bleeding volume, postoperative exhaust time and length of hospital stay were compared .Wexner constipation score and gastrointestinal quality of life index ,ab-dominal pain ,frequency score and emptying time of ileocecus before and after 6 months and 12 months between the two groups were com-pared.Results There was no statistically significant difference in the operation time ,intraoperative bleeding volume ,postoperative exhaust time and hospitalization time of two groups (P>0.05).Wexner constipation score,abdominal pain,frequency score of the two groups after 6 months and 12 months decreased significantly (P<0.05,P<0.01),of which the group B was lower than that of the group A (P<0.05,P<0.01). The gastrointestinal quality of life index after operation significantly increased (P<0.01),of which the group B was higher than that of the group A (P<0.01).Ileocecal emptying time of the group B 12 months after operation was shorter than that of the group A (P<0.01),the differences were statistically significant .Conclusion Subtotal colectomy with antiperistaltic cecorectal anastomosis is an effective method to treat STC,which can reduce the length of preserved ileocecum and improve the prognosis of patients .

OBJECTIVE: To investigate the expression of Bax and PHF20 in laryngeal squamous cell carcinoma (LSCC)and to discuss their relevance and the roles in carcinogenesis and development in LSCC. METHOD: The expressions of Bax and PHF20 in the LSCC tissues and normal mucosa tissues adjacent to carcinoma were detected by SP immunohistochemistry assay. The relationship between the expressions of Bax and PHF20 and the clinicopathological characteristics including clinical stage, pathological type, histological grade and lymph node metastasis in LSCC were analyzed according to the clinical data. RESULT: (1) The expressions of Bax and PHF20 were both significantly lower in the LSCC tissue than that in the normal laryngeal tissue (P < 0.01). (2) In clinical stage grouping, there were no statistical differences of the quantity and positive rate of Bax and PHF20 expressions among supraglottic, glottic and subglottic LSCC (P > 0.05). In histological differentiation grouping, the quantity and positive rate of Bax and PHF20 expressions decreased significantly in poorly differentiated LSCC compared with the well and moderately differentiated LSCCs (P < 0.01, P < 0.05, respectively). In T stage grouping, the quantity and positive rate of Bax and PHF20 expressions were both significantly higher in T1 + T2 compared with T3 +T4 (both P < 0.01). In addition, the quantity and positive rate of Bax and PHF20 expressions were both significantly higher in LSCC with lymph node metastasis compared to that without lymph node metastasis (both P < 0.01). CONCLUSION The lack of Bax and PHF20 might contribute to the carcinogenesis and development in LSCC. The positive expression of Bax and PHF20 maybe relative to T term degree, differentiation degree and lymphamatic metastasis of LSCC.
Antigens, Neoplasm ; metabolism ; Biomarkers, Tumor ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; Head and Neck Neoplasms ; metabolism ; Humans ; Immunohistochemistry ; Laryngeal Neoplasms ; metabolism ; Lymphatic Metastasis ; Prognosis ; Squamous Cell Carcinoma of Head and Neck ; bcl-2-Associated X Protein ; metabolism

Objective To understand the species,species distribution,the dominant species and their interspecies interaction of chigger mites on Eothenomys miletus(a dominant species of rats)in Yunnan.Method The rats were captured with mouse traps in 16 counties(or cities)during 2000-2004.All mites on the surface of two auricles of the hosts were collected and identified.The patch index(m*/m)and the coefficient of association(V)were adopted to judge the spatial distribution patterns and interspecies interaction of the dominant chigger mite species among different individuals of the rats(E.miletus).Results 1157 individuals of E.miletus were captured from 16 counties(citys).37613 chigger mites(belonging to 3 subfamily,9 genus and 80 species)were collected from the auricles(body surface)of 1157 rat hosts with a high “overall mite infestation rate”(68.2%)and “overall mite index”(32.5).Six species of mites were found dominant on E.miletus:Leptotrombidium scutellare,Leptotrombidium sinicum,Helenicula simena,Leptotrombidium eothenomydis,Herpetacarus hastoclavus and Leptotrombidium hiemalis.The distribution of the chigger mites among different individuals of E.miletus showed an aggregation pattern.Both positive and negative association existed between each two dominant species of chigger mites.Conclusion The species composition of chigger mites on Eothenomys miletus is complex with abundant individuals,which reflects a high species diversity of the mites.The main species of chigger mites tend to an aggregation on the body surface of E.miletus.