[摘 要] 目的：探究半乳糖凝集素-9（Gal-9）对胰腺癌细胞的侵袭与迁移功能的促进作用及其机制。方法：基于公开单细胞转录组数据分析胰腺癌中免疫检查点分子的表达谱，结合Western blotting和ELISA验证Gal-9在胰腺癌细胞及临床相关模型中的表达与分泌特征。建立C57BL/6小鼠胰腺原位移植瘤模型，通过免疫组化、免疫荧光染色和流式细胞术分析Gal-9对肿瘤细胞增殖及其对瘤内CD8+ T细胞浸润与功能状态的影响。Gal-9重组蛋白及肿瘤细胞条件培养基（CM）处理肿瘤相关成纤维细胞（CAF），Western blotting、RT-qPCR和免疫荧光检测CAF中Ⅰ型胶原表达变化。免疫共沉淀、免疫荧光染色和Western blotting分析Gal-9与CD44的相互作用及JAK2/STAT3通路相关蛋白的表达。构建过表达Gal-9胰腺癌细胞与CAF共移植的裸鼠皮下荷瘤模型，结合天狼星红和马松染色评估小鼠肿瘤组织中胶原沉积情况。脱细胞外基质法提取CAF中沉积的细胞外基质（ECM）蛋白；通过细胞划痕、Transwell迁移及侵袭实验，检测CAF来源Ⅰ型胶原对胰腺癌细胞侵袭、迁移功能的影响，并通过siRNA干扰Gal-9表达进行功能反向验证。结果：与经典免疫检查点分子PD-L1和CTLA-4相比，Gal-9在胰腺癌中显著高表达（P < 0.000 1）。体内实验表明，Gal-9促进肿瘤生长（P < 0.001），上调Ki-67表达，同时减少肿瘤内CD8⁺ T细胞浸润并诱导肿瘤组织中TIM-3⁺细胞比例升高，Granzyme B、IFN-γ及TNF-α表达下降（P < 0.05）。体外实验显示，外源性Gal-9及肿瘤细胞来源的CM均可上调CAF中Ⅰ型胶原表达（P < 0.001）。机制研究表明，在CAF中抗Gal-9抗体免疫沉淀复合物中检测到CD44，且JAK2/STAT3蛋白表达水平增加。CAF来源的Ⅰ型胶原处理胰腺癌细胞后，肿瘤细胞的迁移和侵袭数量增加。相反，肿瘤细胞中敲低Gal-9表达后，CAF来源Ⅰ型胶原水平下降，同时肿瘤细胞侵袭和迁移数量下降。结论：胰腺癌来源的Gal-9具有双重作用：一方面，通过促进肿瘤细胞增殖和诱导CD8+ T细胞耗竭促进肿瘤进展；另一方面，通过结合CAF上的CD44受体，激活JAK2/STAT3信号通路，刺激CAF合成并分泌Ⅰ型胶原，进而增强胰腺癌细胞的侵袭迁移能力。

Objective: To study the effect of metformin sensitizing pancreatic cancer cells with radiotherapy, with a focus on elucidating the underlying mechanisms of radiotherapy resistance. In particular, the role of the PERK/P-eIF2/ATF4 signaling pathway in mediating these effects was preliminarily explored. Methods : Pancreatic cancer cell lines(PANC-1 and PANC-2) were categorized into control, radiotherapy, and drug treatment groups. Following the respective treatments, cell proliferation inhibition was assessed using the CCK-8 assay, colony formation assays, and cell death staining. Senescence was quantified by β-galactosidase(SA-β-Gal) staining. The expression of cell cycle regulators(P21, P16, γ-H2AX), apoptosis markers(Bax, Bcl-2, Cleaved caspase-3), and pathway-related proteins(PERK, P-eIF2, ATF4) was evaluated by Western blot and immunofluorescence. To further investigate the role of the PERK/P-eIF2/ATF4 axis in metformin-mediated modulation of pancreatic cancer cell senescence and radiosensitization, selective inhibitors(GSK2606414) and agonists(MK-28) of PERK were employed. Results : Radiotherapy markedly upregulated senescence-associated markers(P21, P16, γ-H2AX, and β-galactosidase activity) in pancreatic cancer cells. Senescent cells exhibited enhanced proliferative activity and increased tumor volume both in vitro and in vivo. Metformin mitigated radiotherapy-induced senescence by reducing the expression of senescence markers and significantly suppressing the clonogenic and proliferative capacity of treated cells. Mechanistically, radiotherapy activated the PERK signaling pathway, leading to increased expression of PERK, P-eIF2, and ATF4, thereby driving cellular senescence. Pharmacological inhibition of PERK reduced β-galactosidase activity, while PERK activation further promoted the expression of senescence-associated proteins—an effect that was reversed by metformin. Conclusion Metformin inhibits the activation of the PERK/P-eIF2/ATF4 signaling pathway in pancreatic cancer cells following radiotherapy, thereby delaying cellular senescence and reducing the associated radiotherapy resistance of senescent cells. This modulation contributes to the sensitization of pancreatic cancer cells to radiotherapy.

Through consulting the ancient herbs， medical books and modern literature， this paper made textual research on the name， origin， producing area， quality evaluation， collection and processing of medicinal materials of Sang （Mori Folium， Mori Cortex， Mori Ramulus， Mori Fructus） in famous classical formulas， in order to provide a basis for the development of famous classical formulas containing medicinal materials of Sang. According to the research， Mori Folium and Mori Cortex were first used as medicines in Shengnong Bencaojing ， Mori Ramulus was first used as medicine in Jinxiaofang， and Mori Fructus was first used as medicine in Xinxiu Bencao. Before the Tang dynasty， there were Nyusang and Shansang. Since Tang dynasty， there were many sources of medicinal materials of Sang， including Baisang （Morus alba）， Jisang （M. australis）， Shansang （M. mongolica）， etc. According to textual research， the mainstream varieties were M. australis， M. alba and their cultivated varieties. In modern times， according to the relevant information and the Chinese Pharmacopoeia， M. alba is the original base. In ancient times， the origin of mulberry changed with the development of sericulture， mulberry has been widely planted since the Song dynasty. In the Ming and Qing dynasties， mulberry has been planted most in Jiangsu and Zhejiang. In modern times， they are mainly produced in Jiangsu， Zhejiang， Anhui， Hunan and other places. In recent years， due to the related policies and strategies such as "moving silkworms from east to west"， the center of silkworm breeding has gradually transferred to the west. As for the quality evaluation and harvesting and processing of mulberry medicinal materials， Most of the ancient and modern records of Mori Folium are the same. They are harvested after frost， and dried after removing impurities. The quality is better when the leaves are large and thick， yellowish green， holding prickly hands and undergoing frost. The harvesting period of Mori Cortex is slightly different in ancient and modern records. Ancient books record that it can be harvested all the year round， but in modern times， it is mostly harvested from late autumn to the next spring. The processing methods include removing soil and fibrous roots， scraping off yellow-brown rough skin， peeling off white skin and drying in the sun. The quality is better when they are white， thick， flexible， free of rough skin and full of powder. There are few records about the collection， processing and quality evaluation of Mori Ramulus and Mori Fructus in ancient Chinese herbal books. According to modern literature， Mori Ramulus is usually collected in late spring and early summer， with leaves removed， slightly dried， sliced while fresh， and dried in the sun. The best quality of Mori Ramulus is fine and tender with the yellow and white section. Mori Fructus is harvested from April to June when the fruit turns red， and dried in the sun， or slightly steamed and dried in the sun， and it is better to be big， dark purple， oily and thick. There are many processing methods of mulberry medicinal materials. Ancient books record stir frying， baking， burning and steaming of Mori Folium， in modern times， there is honey-roasted method， but most of them are used as raw products. In ancient materia medica， Mori Cortex has firing method， baking method， stir-frying method， honey-fried method， etc. In modern times， there are stir-fried and honey-fried methods， and most of them are used as raw products. Ancient books record that Mori Ramulus has cutting and frying methods， while modern ones have cutting， frying， wine-processed and bran-processed methods. Processing methods of Mori Fructus are consistent in ancient and modern times， and they are mostly dried after being cleaned or steamed. Based on the research results， it is suggested that M. alba should be selected as mulberry medicinal materials in the famous classical formulas， and appropriate medicinal parts and processing methods can be selected according to the indications of the famous classical formulas.

In this paper， the name， origin， producing area and other aspects of Menthae Haplocalycis Herba in the famous classical formulas were carried out by consulting herbal literature， medical books， prescription books in the past dynasties and related modern documents. Through the textual research， it can be seen that the name of Bohe was used as the correct name in the mainstream of the past dynasties， and there were still multiple synonyms， most of which originated from the false transmission of dialectal accent， producing area and efficacy. There are many varieties recorded in the literature of the past dynasties such as Bohe， Longnao Bohe， Hubohe and Shibohe. According to the textual research， Bohe， Longnao Bohe and Yebohe are consistent with Mentha haplocalyx， whcih is the mainstream variety. Longnao Bohe is named for its form of producing area， Shibohe is Mosla chinensis， Daye Bohe is Agastache rugosa， and Nanbohe is M. crispata. Menthae Haplocalycis Herba has been widely planted since Tang dynasty. It was mainly grown in Jiangsu， Zhejiang， Jiangxi and Sichuan in Ming and Qing dynasties， and Jiangsu is the genuine production area. Its quality is best if it has dry body， no roots， many leaves， green color and strong smell. In ancient times， the stems and leaves of Menthae Haplocalycis Herba were often picked and dried in summer and autumn， which is basically the same as the records of modern times when the stems and leaves are luxuriant in summer and autumn， or when the flowers bloom to three rounds， they are picked in sunny days and cut in different times， and then dried in the sun or in the shade， and the raw products was often used as medicine in ancient and modern times. Before the Song dynasty， Menthae Haplocalycis Herba was recorded as pungent and warm. Until the Song dynasty， it was written as “extremely cool” in Lyuchanyan Bencao. It may have been thought in the early stage that it was similar to several warm herbs， such as Perilla frutescens， Stachys japonica， Elsholtzia ciliata and M. chinensis in appearance， all of which have the function of Xinsan， so it was recorded as warm. Since the Qing dynasty， Menthae Haplocalycis Herba has been recorded as cool property in the mainstream materia medica， Menthae Haplocalycis Herba recorded as pungent and cool in the 2020 edition of Chinese Pharmacopoeia， and its effect is to dissipate wind heat， clear the head， relieve the pharynx and so on， the records of efficacy in ancient and modern times are basically the same. Based on the research results， it is suggested that raw products of M. haplocalyx should be selected when developing the famous classical formulas containing Menthae Haplocalycis Herba.

Objective:The aims of this study were to develop a simple and easy-to-use scale, named Methamphetamine Use Disorder (MUD) Severity Scale (MUDSS), and to complete the reliability and validity tests, so as to provide a new tool for the measurement of substance use.Methods:Participants with MUD ( n=286) were recruited from 2 compulsory isolated detoxification centers in Changsha, Hunan Province from April to September 2019. The item pool of MUDSS ( n=19) was obtained according to diagnostic items of stimulant use disorder in the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5) and the opinions of experts in the field of addiction. The scale was applied to participants with MUD. Items of the scale were first screened according to the results of item analysis, and then the formal scale was formed. Exploratory factor analysis was used to construct the scale dimension, and the reliability of the total scale and each dimension were tested by Cronbach α coefficient. Pearson correlation analysis was used to verify criterion validity and DSM-5 was used as criterion. Results:The effective information of 257 subjects was collected. According to the results of item analysis, the fourth item from the preliminary MUDSS was deleted, and the remaining 18 items were included in the formal scale, including four dimensions: adverse consequences, tolerance, priority, and craving. The internal consistency coefficient of the total scale and the four dimensions ranged from 0.646 to 0.909, and the test-retest reliability ranged from 0.592 to 0.724. The correlation between MUDSS total score and DSM-5 was 0.654.Conclusions:The reliability and validity of MUDSS are good and have reached the standard of psychometrics, therefore it can be further applied in a larger sample of subjects with MUD.

Objective:The aims of this study were to develop a simple and easy-to-use scale, named Methamphetamine Use Disorder (MUD) Severity Scale (MUDSS), and to complete the reliability and validity tests, so as to provide a new tool for the measurement of substance use.Methods:Participants with MUD ( n=286) were recruited from 2 compulsory isolated detoxification centers in Changsha, Hunan Province from April to September 2019. The item pool of MUDSS ( n=19) was obtained according to diagnostic items of stimulant use disorder in the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5) and the opinions of experts in the field of addiction. The scale was applied to participants with MUD. Items of the scale were first screened according to the results of item analysis, and then the formal scale was formed. Exploratory factor analysis was used to construct the scale dimension, and the reliability of the total scale and each dimension were tested by Cronbach α coefficient. Pearson correlation analysis was used to verify criterion validity and DSM-5 was used as criterion. Results:The effective information of 257 subjects was collected. According to the results of item analysis, the fourth item from the preliminary MUDSS was deleted, and the remaining 18 items were included in the formal scale, including four dimensions: adverse consequences, tolerance, priority, and craving. The internal consistency coefficient of the total scale and the four dimensions ranged from 0.646 to 0.909, and the test-retest reliability ranged from 0.592 to 0.724. The correlation between MUDSS total score and DSM-5 was 0.654.Conclusions:The reliability and validity of MUDSS are good and have reached the standard of psychometrics, therefore it can be further applied in a larger sample of subjects with MUD.

Rubi Fructus is a commonly used traditional Chinese medicine. The origin of Rubi Fructus is the dried fruit of Rubus chingii, a plant of the family Rosaceae, according to the 2015 edition of Chinese pharmacopoeia. There are some differences in the plant origin of Rubi Fructus in ancient herbal literature, to trace back its sources, we conducted a textual research on its origin, producing areas, quality evaluation, processing and concocting, properties, tastes and efficacy etc. based on the records of ancient herbal literatures and combined with plant morphology and related investigation. RESULTS:: showed that the variety of Rubi Fructus was more complex among ancient herbal literature, including R. coreanus, R. hirsutus, R. corchorifolius, R. foliolosus and other mixed varieties. Most scholars believe that the R. chingii has not been recorded in ancient herbal literature, while R. chingii was recorded as early as the Ming Dynasty in Compendium of materia medica through our textual research. Ancient Chinese herbs recorded that Rubi Fructus was mostly produced in Hubei, Shandong, Shanxi and Jiangsu provinces, while R. chingii mainly produced in Anhui, Jiangsu, Zhejiang, Jiangxi, Fujian and other provinces nowadays. Also, it was recorded that Rubi Fructus harvested in wheat field during May were the best. Besides, R. chingii with big, full, grain integrate, firm, yellow and green color, sour taste and impurity free possess the best quality in the contemporary. The ancient records of processing and concocting, properties, tastes and efficacy were basically the same as modern ones.These results provide the basis for the correct utilization and further development of Rubi Fructus.
China ; Drugs, Chinese Herbal ; Fruit ; Humans ; Materia Medica ; Medicine, Chinese Traditional ; Rubus

Visci Herba， a commonly used Chinese medicinal， was often mistaken as Taxilli Herba in ancient Chinese materia medica. The two Chinese medicinals have often been confused even in present clinical practice， and their origins are difficult to be identified. Hence， it is necessary to carry out systematic and in-depth textual research. This paper explored the origin， producing area， quality evaluation， harvesting and processing， property， flavor， and efficacy of Visci Herba based on ancient Chinese materia medica of the past dynasties and modern plant morphology， so as to provide evidence for the development and utilization of Visci Herba. The findings demonstrated that Visci Herba was mostly recorded as Taxilli Herba until the name of Visci Herba appeared in the Tang Dynasty. The records of the two Chinese medicinals could be traced back to the Song Dynasty. Visci Herba and Taxilli Herba were officially listed as two different Chinese medicinals in the 1977 edition Chinese Pharmacopoeia for the first time， where the origin of Visci Herba was determined to be Viscum coloratum（Komar.）Nakai. According to the ancient Chinese materia medica， V. coloratum was mainly distributed in Lingbao， Henan province and Xuzhou， Jiangsu province and now it mostly grows in Northeast China and North China. In ancient times， Visci Herba with deep yellow cross-section and sticky fruit juice on the tree was preferred， which was often harvested on 3， March in spring， dried in the shade， grinded together with the roots， branches， stems， and leaves using the copper file， and preserved away from fire. By contrast， it is now usually harvested from winter to the next spring and then cut into sections for drying after the removal of thick stems， or dried after being steamed. As described in ancient Chinese materia medica， Visci Herba， bitter and sweet in flavor， neutral in property， possessed the effects of dispelling rheumatism， tonifying kidney， strengthening sinews and bones， benefiting blood vessels， and preventing abortion， basically consistent with its modern functions of dispelling rheumatism， tonifying liver and kidney， strengthening sinews and bones， and preventing abortion.

Background: Our previous studies have shown that regulatory factor X5 (RFX5), a classical transcription regulator of MHCII genes, was obviously overexpressed in hepatocellular carcinoma (HCC) tumors. However, the role of RFX5 in the carcinogenesis and progress of HCC remains unknown. This study aimed to reveal its biological significance and the underlying mechanism in HCC. Methods: RFX5 mRNA expression level and copy number variation in HCC tumors and cell lines were determined by analyzing deposited data sets in the Cancer Genome Atlas and Gene Expression Omnibus database. The biological significance of RFX5 in HCC was investigated by monitoring the colony formation and subcutaneous tumor growth capacity when RFX5 was silenced with lentiviral short hairpin RNA and CRISPR/Cas9 system in HCC cell lines. The downstream gene transcriptionally activated by RFX5 in HCC cells was determined by chromatin immunoprecipitation and luciferase reporter assay. The involvement of tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein theta (YWHAQ) in HCC development was further determined by performing colony formation rescue assay and subcutaneous tumor growth rescue experiment. The association of YWHAQ with recurrence-free survival of patients with HCC was assessed by Kaplan-Meier analysis. Moreover, apoptosis level and the protein level of p53 pathway were determined to reveal the mechanism of RFX5 in driving HCC development. Results: RFX5 was amplified and highly overexpressed in HCC tumor tissues compared with the corresponding non-tumor tissues. The mRNA expression level of RFX5 was significantly correlated with its DNA copy number (r = 0.4, P < 0.001). Functional study demonstrated that RFX5 was required for both clonogenic forming in vitro and subcutaneous tumor growth in vivo of HCC cells. Further study identified YWHAQ, namely 14-3-3 tau, as a key downstream transcriptional target gene of RFX5, which was tightly regulated by RFX5 in HCC. Moreover, overexpression of YWHAQ largely rescued the clonogenic growth of HCC cells that was suppressed by RFX5 knockdown. In addition, overexpression of YWHAQ in primary tumor was linked to poor prognosis of patients with HCC. These results demonstrated that YWHAQ was a downstream effector of RFX5 in HCC. Notably, RFX5-YWHAQ pathway could protect cells from apoptosis by suppressing the p53 and Bax in HCC. Conclusion RFX5 is a putative HCC driver gene that plays an important role in the development and progression of HCC by transactivating YWHAQ and suppressing apoptosis.

Objective To evaluate the clinical efficacy and safety of external use of recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) gel on deep partial-thickness burn wounds.Methods Sixty-eight hospitals in our country including our unit performed a phase Ⅳ clinical trial for rhGM-CSF gel in patients (conforming to the study criteria) with deep partial-thickness burn wounds from November 2010 to July 2012.Multicenter,randomized,positive-homogenous-controlled,and open trial method was used in the trial,and patients from 10 hospitals were grouped into the positive-homogenous-controlled trial,while patients from the other 58 hospitals were grouped into open trial.(1) Controlled trial.Patients were divided into rhGM-CSF group and conventional treatment group (CT) with the ratio of 1:1 according to the stratified randomization method.Wounds of patients in rhGM-CSF group were coated with rhGM-CSF gel,and wounds of patients in group CT were covered by gauze with iodophor.Scores of wound exudate and wound edge response before treatment and on treatment day (TD) 2,4,8,10,14,20,and 28 were conventionally evaluated.Wound healing rates on TD 8,10,14,20,and 28 were calculated.Complete wound healing time and overall efficiency including cure,excellence,progress,and invalid situation on TD 28 were recorded.Safety indexes including vital signs and laboratory test indexes before and during treatment,and adverse reactions during treatment were observed.(2) Open trial.Wounds of patients in this trail were all coated with rhGM-CSF gel.Complete wound healing time,overall efficiency,and safety indexes of patients were recorded as in controlled trial.Data were processed with CMH-x2 test,Fisher's exact test,signed rank sum test,paired t test,Log-Rank test,and Wilcoxon rank sum test.Results (1) Controlled trail.A total of 366 patients from 10 hospitals were included in this trial,and 358 cases with 177 cases in rhGM-CSF group and 181 cases in group CT finished the trial.There were no statistically significant differences in gender,age,injury characteristics,and combined medication situation between patients in two groups (x2 =1.510,with t values from 0.458 to 0.820,P values above 0.05).Scores of wound exudate of patients in two groups were similar before treatment and on TD 2,20,and 28 (t =0.420,with Z values from 0.735 to 1.939,P values above 0.05).Scores of wound exudate of patients in rhGM-CSF group were significantly lower than those in group CT on TD 4,8,10,and 14 (with Z values from 2.054 to 2.580,P values below 0.05).Scores of wound edge response of patients in two groups were similar before treatment and on each TD (t =0.340,with Z values from-1.147 to 1.874,P values above 0.05).Wound healing rate of patients in rhGM-CSF group was significantly higher than that in group CT on each TD (with Z values from 2.630 to 5.235,P values below 0.01).The complete wound healing time of patients in rhGM-CSF group was (16.93 ±0.40) d,which was significantly shorter than that in group CT [(19.88 ± 0.41) d,x2 =26.732,P ＜0.001].At last,171 (96.61％) patients were completely cured in rhGM-CSF group,while excellence,progress,and invalid results were achieved in 3 (1.69％),1 (0.56％),and 2 (1.13％) patients,respectively.Whereas,161 (88.95％) patients were completely cured in group CT,while excellence,progress,and invalid results were achieved in 1 1 (6.08％),5 (2.76％),and 4 (2.12％) patients,respectively.Total efficacy of patients in rhGM-CSF group was significantly higher than that in group CT (x2 =5.784,P ＜ 0.05).Levels of vital signs and laboratory test indexes of patients in two groups before and during treatment were similar.There were no statistically significant differences in adverse reaction or drug-related adverse reaction between patients in two groups during treatment (with P values above 0.05).(2) Open trial.A total of 2 380 patients were enrolled in,and 2 329 patients finished the trial.The complete wound healing time of patients was (16.28 ± 0.10)d.At last,2 257 (96.91％) patients were totally cured,while excellence,progress,and invalid results were achieved in 36 (1.55％),16 (0.69％),and 20 (0.86％) patients,respectively.Vital signs and laboratory test indexes of patients before and during treatment were similar.The drug-related adverse reaction was observed in 44 patients (1.89％).Conclusions External use of rhGM-CSF gel on deep partial-thickness burn wounds can promote wound healing and is safe for clinical use.