1.Herbal Textual Research on Stemonae Radix in Famous Classical Formulas
Gang XU ; Li AN ; Xiaomei WANG ; Erhuan WANG ; Yichen YANG ; Cunde MA ; Yang YANG ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):241-251
This article systematically reviews and verifies the historical evolution of Stemonae Radix from the aspects of name, origin, harvesting and processing, quality and others by consulting ancient and modern literature, in order to provide reference for the development and utilization of famous classical formulas containing this medicinal herb. Stemonae Radix has a long history of application, and it derives its name from its distinctive growth pattern, featuring clusters of ten to several dozen underground tuberous roots. This morphology resembles that of certain plants in the genus Asparagus, leading to historical instances where tuberous roots from genus Asparagus were mistakenly used as Stemonae Radix. After the research, it can be concluded that Stemonae Radix was first recorded in Mingyi Bielu, and throughout history, Baidu has been recognized as its official name, though it also bears alternative names such as Baibing, Pofucao and Ye Tianmendong. The mainstream sources used throughout history have been the dried tuberous roots of Stemona sessilifolia, S. japonica or S. tuberosa from the family Stemonaceae. This aligns with the 2025 edition of Pharmacopoeia of the People's Republic of China(hereinafter referred to as Chinese Pharmacopoeia). Additionally, Asparagus filicinus and A. officinalis from the genus Asparagus are common sources of confusion with Stemonae Radix. The three primitive plants are mainly distributed in the Yangtze River basin and southern China, exhibiting a wide distribution. Historically, wild harvesting was predominant, but cultivation is now established. In ancient times, the harvesting time was mostly in the second, third, and eighth lunar months, when roots were harvested and dried. Nowadays, it is more common to pick and excavate in the spring and autumn seasons. After excavation, the roots are washed, fibrous roots removed, briefly blanched in boiling water or steamed until no white core remains, and then sun-dried or oven-dried. In ancient times, the processing of Stemonae Radix primarily involved roasting(stir-frying), wine roasting, or raw materials. Modern mainstream processing specifications include two types of raw and honey-roasted products. In terms of quality evaluation of the medicinal materials, ancient criteria of "preferring plump and moist roots" align with modern requirement favoring "thick, robust stems with firm texture". Evaluating quality with authenticity, since the Song dynasty, it has been highly praised to produce in Chuzhou and Hengyang as the best. It was an ancient method of fixing the production area to stabilize the medicinal origin, reflecting the ancient recognition of the therapeutic efficacy of plants belonging to the genus Stemona. The main functions of Stemonae Radix remain consistent throughout history, including relieving coughs, eliminating phlegm and parasites. Based on the research results, it is recommended that when developing famous classical formulas containing the medicinal material Stemonae Radix, the botanical source specified in the 2025 edition of Chinese Pharmacopoeia should be selected. The specific species can be determined according to the distribution of resources and the main production areas, and the origin and corresponding botanical source should be fixed. Processing methods should be chosen based on the prescription requirements. It is recommended to use raw products without specified requirements.
2.Herbal Textual Research on Stemonae Radix in Famous Classical Formulas
Gang XU ; Li AN ; Xiaomei WANG ; Erhuan WANG ; Yichen YANG ; Cunde MA ; Yang YANG ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):241-251
This article systematically reviews and verifies the historical evolution of Stemonae Radix from the aspects of name, origin, harvesting and processing, quality and others by consulting ancient and modern literature, in order to provide reference for the development and utilization of famous classical formulas containing this medicinal herb. Stemonae Radix has a long history of application, and it derives its name from its distinctive growth pattern, featuring clusters of ten to several dozen underground tuberous roots. This morphology resembles that of certain plants in the genus Asparagus, leading to historical instances where tuberous roots from genus Asparagus were mistakenly used as Stemonae Radix. After the research, it can be concluded that Stemonae Radix was first recorded in Mingyi Bielu, and throughout history, Baidu has been recognized as its official name, though it also bears alternative names such as Baibing, Pofucao and Ye Tianmendong. The mainstream sources used throughout history have been the dried tuberous roots of Stemona sessilifolia, S. japonica or S. tuberosa from the family Stemonaceae. This aligns with the 2025 edition of Pharmacopoeia of the People's Republic of China(hereinafter referred to as Chinese Pharmacopoeia). Additionally, Asparagus filicinus and A. officinalis from the genus Asparagus are common sources of confusion with Stemonae Radix. The three primitive plants are mainly distributed in the Yangtze River basin and southern China, exhibiting a wide distribution. Historically, wild harvesting was predominant, but cultivation is now established. In ancient times, the harvesting time was mostly in the second, third, and eighth lunar months, when roots were harvested and dried. Nowadays, it is more common to pick and excavate in the spring and autumn seasons. After excavation, the roots are washed, fibrous roots removed, briefly blanched in boiling water or steamed until no white core remains, and then sun-dried or oven-dried. In ancient times, the processing of Stemonae Radix primarily involved roasting(stir-frying), wine roasting, or raw materials. Modern mainstream processing specifications include two types of raw and honey-roasted products. In terms of quality evaluation of the medicinal materials, ancient criteria of "preferring plump and moist roots" align with modern requirement favoring "thick, robust stems with firm texture". Evaluating quality with authenticity, since the Song dynasty, it has been highly praised to produce in Chuzhou and Hengyang as the best. It was an ancient method of fixing the production area to stabilize the medicinal origin, reflecting the ancient recognition of the therapeutic efficacy of plants belonging to the genus Stemona. The main functions of Stemonae Radix remain consistent throughout history, including relieving coughs, eliminating phlegm and parasites. Based on the research results, it is recommended that when developing famous classical formulas containing the medicinal material Stemonae Radix, the botanical source specified in the 2025 edition of Chinese Pharmacopoeia should be selected. The specific species can be determined according to the distribution of resources and the main production areas, and the origin and corresponding botanical source should be fixed. Processing methods should be chosen based on the prescription requirements. It is recommended to use raw products without specified requirements.
3.Hypericin reduces myocardial ischemia-reperfusion injury in rats by modulating AMPK/Nrf2/HO-1 signaling pathway
Huihui LI ; ANWER GULINIGAER ; Xiaofeng GAO ; Gang WANG ; Yali BAO ; Tian ZHANG ; AINIWAR DINA ; Can LING ; Zhan SUN
Chinese Journal of Pathophysiology 2024;40(10):1882-1890
AIM:To investigate the effect of hypericin(Hyp)on the heart of rats with myocardial ischemia-re-perfusion injury(MIRI),and to explore its mechanism.METHODS:Thirty SPF male SD rats were divided into 5 groups:sham group,MIRI group,low-dose Hyp(L-Hyp)group(MIRI+L-Hyp group),high-dose Hyp(H-Hyp)group(MIRI+H-Hyp group),and positive control trimetazidine(TMZ)group(MIRI+TMZ group),with 6 rats in each group.Apart from the sham group,the MIRI model was established by ligating the anterior descending branch of the left coronary artery and then recanalizing it in the remaining four groups of rats.The success of the modeling was determined by monitor-ing the electrocardiogram.We assessed the cardiac function in rats using echocardiography.TTC staining was employed to measure the area of myocardial infarction in rats,and HE staining was utilized to observe the morphological traits of rat myocardium.We assayed the activities of lactate dehydrogenase(LDH)and superoxide dismutase(SOD)and the levels of malondialdehyde(MDA)in rat serum using biochemical kits.ELISA kits were applied to assess the contents of cardiac troponin I(cTnI)and reactive oxygen species(ROS)in rat serum.Western blot analysis was perfomed to measure the pro-tein expression levels of AMPK,p-AMPK,Nrf2,and HO-1 in rat myocardial tissues.RESULTS:The rats in MIRI group exhibited increased myocardial tissue injury,larger myocardial infarction areas,decreased left ventricular ejection fraction(LVEF),and reduced left ventricular fractional shortening(LVFS)compared with sham group,as shown by echo-cardiography.Additionally,there were increases in LDH activity,cTnI,MDA and ROS levels,along with significant de-creases in SOD activity,and p-AMPK,Nrf2 and HO-1 protein levels(P<0.05).Compared with MIRI group,the rats in MIRI+L-Hyp,MIRI+H-Hyp and MIRI+TMZ groups showed decreased myocardial histopathological damage and reduced myocardial tissue infarction area,increased LVEF and LVFS,and lowered serum levels of LDH activity,cTnI,MDA and ROS,while SOD activity,p-AMPK,Nrf2 and HO-1 protein levels were elevated(P<0.05).CONCLUSION:Hypericin attenuates myocardial ischemia-reperfusion injury in rats,possibly by modulating the AMPK/Nrf2/HO-1 signaling path-way.
4.A method for determination of fluoride in urine by automatic potentiometric titrator
Shudong XU ; Gang DING ; Ji'an XIE ; Ziyue ZHAN ; Qi ZHANG ; Xian XU ; Weidong LI
Chinese Journal of Endemiology 2024;43(7):598-602
Objective:To establish an automatic potentiometric method for determination of fluoride in urine.Methods:The fluorine level in human urine was determined by an automatic potentiometric titrator. Methodological experimental analysis of the linear range, detection limit, precision, and accuracy (quality control sample testing and actual sample spiking recovery experiment) of the method were carried out. The determination results of 33 urine samples were compared with those of the national health industry standard Determination of Fluoride in Urine - Ion Selective Electrode Method (WS/T 89-2015).Results:The linear range of the standard curve of urine fluoride in the automatic potentiometric determination method was 0.1 - 10.0 mg/L. The range of linear correlation coefficient ( r) was 0.999 6 to 0.999 9. The minimum detection limit of urine fluoride was 0.01 mg/L (sampling volume was 10.0 ml). The relative standard deviation ( RSD) range for precision experiments was 0.00% to 3.45%. The quality control samples of the national urine fluoride external quality control assessment in 2022 were determined, and the results were within the range of the consensus value. The spiked recovery experiment was carried out, the recovery rate ranged from 100.0% to 104.4%, with a total average recovery rate of 102.1%. The method comparison experiment showed that there was no statistically significant difference between the test results of automatic potentiometric method and the standard method recommended by the national urine fluoride health industry ( t = - 0.90, P = 0.375). Conclusions:A method has been successfully established, achieving automation of the reagent addition and detection process of urine fluoride samples after sampling. It has the characteristics of wide linear range, low detection limit, good precision, and high accuracy, and is suitable for rapid and automatic determination fluoride level of large quantities of urine samples.
5.Dosimetric study of two-arc and dual-arc techniques in VMAT program for lower mid-thoracic esophageal cancer
Yong-Fu FENG ; Yu-Song LONG ; Jun-Wen TAN ; Xian-Tao HE ; Gang LI ; Zhan-Yu WANG
Chinese Medical Equipment Journal 2024;45(1):62-66
Objective To compare the effects of two arc(TA)and dual arc(DA)techniques on the dose distribution to the planning target volume(PTV)and organs at risk(OAR)in volumetric modulated arc therapy(VMAT)for lower mid-thoracic esophageal cancer.Methods Ten patients with lower mid-thoracic esophageal cancer who received radiation therapy at some hospital from July 2020 to June 2022 were selected retrospectively.A TA radiation therapy plan and a DA radiation therapy plan were developed for each patient using the Ray Arc module of RayStation 4.7.5.4 planning system,and the two kinds of radiation plans were compared in terms of dosimetric parameters including D2,D5,D50,D95,D98,homogeneity index(HI),conformity index(CI),beam-on time and total monitor unit for PTV and lung V5,V10,V20,V30 and Dmean and heartV30,V40 and Dmean and spine cord Dmax for OAR.SPSS 22.0 was used for statistical analysis.Results TA and DA radiation therapy plans had no significant differences in PTV CI,HI,D2,D5,D50,D95 and beam-on time(P>0.05),and DA plan had D98 and total monitor unit higher obviously than those of TA plan(P<0.05).In terms of OARs protection,DA plan had heart V30,V40 and Dmean slightly lower than those of TA plan with non-significantly differences(P>0.05),while lung V5,V30 and Dmean and spine cordDmax significantly lower(P<0.05).Conclusion DA technique gains advantages over TA technique in PTV dose distribution and dose to OAR,and the involvement of DA technique in preparing the VMAT plan for esophageal cancer contributes to enhancing the treatment efficacy.[Chinese Medical Equipment Journal,2024,45(1):62-66]
6.Distribution of urinary tract pathogens and susceptibility to nenofloxacin and levofolxacin in patients with community-acquired urinary tract infection
Zhen DU ; Shan CHEN ; Liang CUI ; Huiling WU ; Zhan GAO ; Jin YANG ; Gang CUI ; Jiajing WANG ; Tiehuan SHU ; Ninghan FENG ; Ludong QIAO
Chinese Journal of Urology 2024;45(1):24-28
Objective:To investigate the distribution characteristics of urinary tract pathogens in patients with community-acquired urinary tract infection and their sensitivity to nenoxacin and levofloxacin.Methods:This prospective, multicenter clinical trial included patients with community-acquired urinary tract infection who were admitted to urological clinics at 9 clinical research centers from November 2021 to August 2022.Inclusion criteria: Patients aged 18-70 years with community-acquired acute uncomplicated cystitis(AUC), recurrent acute episodes of urinary tract infection(rUTI), and non-febrile complicated urinary tract infection(cUTI) with signs of urinary tract irritation and abnormal elevation of routine white blood cells in urine. Exclusion criteria: ①Patients who received effective antimicrobial therapy within 72 h before enrollment and lasted for more than 24 h. ②Fever (>37.3℃) or symptoms of upper urinary tract infection such as low back pain, tapping pain in the kidney area, etc. ③Indwelling urinary catheter. At the first visit, clean midstream urine samples were taken for bacterial culture, and the distribution characteristics of urinary pathogens of different types of urinary tract infections were analyzed. Extended spectrum β-lactamases (ESBLs) were measured for Gram-negative bacteria. The susceptibility of nenoxacin and levofloxacin to urinary tract pathogens was determined by disk diffusion method. Drug resistance rate, sensitivity rate were analyzed between different disease groups.Results:There were 404 enrolled patients from 9 hospitals, including 364 (90.1%) females and 40 (9.9%) males. A total of 177 strains of pathogenic bacteria were isolated, among which the highest proportion of Escherichia coli was 66.1% (117/177).Klebsiella pneumoniae was followed by 6.8% (12/177) and Streptococcus agalactis 5.1% (9/177). The bacterial spectrum distribution of AUC and rUTI were similar, and the proportions of Escherichia coli were 70.6% (85/119) and 65.9% (29/44), respectively. However, the proportions of Escherichia coli isolated from cUTI patients were only 28.6% (4/14) and Enterococcus faecalis 7.1%(1/14). The overall detection rate of ESBLs in Gram-negative bacteria was 30.9%(43/139). The sensitivity rate of nenoxacin was 74.6%(91/122), and the resistance rate was 25.4%(31/122). The overall sensitivity rate of levofloxacin was 44.9%(70/156) and the resistance rate was 36.5%(57/156). The rate of resistance of urinary tract pathogens to levofloxacin was 48.2% (27/56) in patients with previous urinary tract infection history, and 30.0% (30/100) in patients with no previous urinary tract infection history, the difference was statistically significant( P=0.023).The sensitivity rate of Gram-negative bacteria to nenofloxacin was 70.7% (65/92) and that to levofloxacin was 50.0% (46/92, P<0.001). The sensitivity of Gram-positive bacteria to nenofloxacin was 80.0% (16/20), and that to levofloxacin was 70.0% (14/20, P=0.009). Conclusions:The bacterial profile of out-patient community acquired urinary tract infection varies greatly according to different diseases. The proportion of Escherichia coli in AUC and rUTI patients is higher than that in cUTI. The detection rate of ESBLs in Gram-negative bacteria was lower than the domestic average.Patients with a history of urinary tract infection had a high risk of treatment failure with levofloxacin. The sensitivity of common urinary tract pathogens to nenofloxacin was higher than levofloxacin.
7.Standard for the management of hyperkalemia—whole-process management mode of multi- department cooperation
Zhiming YE ; Jianfang CAI ; Wei CHEN ; Hong CHENG ; Qiang HE ; Rongshan LI ; Xiangmin LI ; Xinxue LIAO ; Zhiguo MAO ; Huijuan MAO ; Ning TAN ; Gang XU ; Hong ZHAN ; Hao ZHANG ; Jian ZHANG ; Xueqing YU
Chinese Journal of Nephrology 2024;40(3):245-254
Hyperkalemia is one of the common ion metabolism disorders in clinical practice. Hyperkalemia is defined as serum potassium higher than 5.0 mmol/L according to the guidelines at home and abroad. Acute severe hyperkalemia can cause serious consequences, such as flaccid paralysis, fatal arrhythmia, and even cardiac arrest. The use of renin-angiotensin- aldosterone system inhibitors, β-blockers and diuretics, low-sodium and high-potassium diets, and the presence of related comorbidities increase the occurrence of hyperkalemia. Hyperkalemia risk exist in all clinical departments, but there is a lack of a standardization in the management of multi- department cooperation in hospital. Therefore, a number of domestic nephrology and cardiology department experts have discussed a management model for multi-department cooperation in hyperkalemia, formulating the management standard on hospital evaluation, early warning, diagnosis and treatment, and process. This can promote each department to more effectively participate in nosocomial hyperkalemia diagnosis and treatment, as well as the long-term management of chronic hyperkalemia, improving the quality of hyperkalemia management in hospital.
8.Analysis of Chinese Medical Syndrome Features of Ischemic Stroke Based on Similarity of Symptoms Subgroup.
Xiao-Qing LIU ; Run-Shun ZHANG ; Xue-Zhong ZHOU ; Hong ZHOU ; Yu-Yao HE ; Shu HAN ; Jing ZHANG ; Zi-Xin SHU ; Xue-Bin ZHANG ; Jing-Hui JI ; Quan ZHONG ; Li-Li ZHANG ; Zi-Jun MOU ; Li-Yun HE ; Lun-Zhong ZHANG ; Jie YANG ; Yan-Jie HU ; Zheng-Guang CHEN ; Xiao-Zhen LI ; Yan TAN ; Zhan-Feng YAN ; Ke-Gang CAO ; Wei MENG ; He ZHAO ; Wei ZHANG ; Li-Qun ZHONG
Chinese journal of integrative medicine 2023;29(5):441-447
OBJECTIVE:
To derive the Chinese medicine (CM) syndrome classification and subgroup syndrome characteristics of ischemic stroke patients.
METHODS:
By extracting the CM clinical electronic medical records (EMRs) of 7,170 hospitalized patients with ischemic stroke from 2016 to 2018 at Weifang Hospital of Traditional Chinese Medicine, Shandong Province, China, a patient similarity network (PSN) was constructed based on the symptomatic phenotype of the patients. Thereafter the efficient community detection method BGLL was used to identify subgroups of patients. Finally, subgroups with a large number of cases were selected to analyze the specific manifestations of clinical symptoms and CM syndromes in each subgroup.
RESULTS:
Seven main subgroups of patients with specific symptom characteristics were identified, including M3, M2, M1, M5, M0, M29 and M4. M3 and M0 subgroups had prominent posterior circulatory symptoms, while M3 was associated with autonomic disorders, and M4 manifested as anxiety; M2 and M4 had motor and motor coordination disorders; M1 had sensory disorders; M5 had more obvious lung infections; M29 had a disorder of consciousness. The specificity of CM syndromes of each subgroup was as follows. M3, M2, M1, M0, M29 and M4 all had the same syndrome as wind phlegm pattern; M3 and M0 both showed hyperactivity of Gan (Liver) yang pattern; M2 and M29 had similar syndromes, which corresponded to intertwined phlegm and blood stasis pattern and phlegm-stasis obstructing meridians pattern, respectively. The manifestations of CM syndromes often appeared in a combination of 2 or more syndrome elements. The most common combination of these 7 subgroups was wind-phlegm. The 7 subgroups of CM syndrome elements were specifically manifested as pathogenic wind, pathogenic phlegm, and deficiency pathogens.
CONCLUSIONS
There were 7 main symptom similarity-based subgroups in ischemic stroke patients, and their specific characteristics were obvious. The main syndromes were wind phlegm pattern and hyperactivity of Gan yang pattern.
Humans
;
Syndrome
;
Ischemic Stroke
;
Medicine, Chinese Traditional
;
Liver
;
Phenotype
9.Effects of miR-143 on the migration and invasion of osteosarcoma cells by regulating MMP-13 expression.
Bin LI ; Zhan-Peng LI ; Zhen-Gang LIAN
China Journal of Orthopaedics and Traumatology 2023;36(11):1075-1080
OBJECTIVE:
To explore the effect of miR-143 regulating matrix metalloproteinase(MMP)-13 expression on migration and invasion of osteosarcoma cells.
METHODS:
The mouse osteosarcoma cell line 143B cells were cultured in 96-well plates, and blank group, negative group, positive group, and intervention group were set up. Then, the blank group did no treatment 50 μg miR-143 mimic was added to positive group, negative group added equal mimic NC (control sequence of miR-143 mimic), the intervention group was added 50 μg miR-143 mimic and 10 μg MMP-13 protein, all groups continued to culture for 3 to 6 hours, and finally the serum was aspirated to treat for half an hour. The protein expressions of miR-143 and MMP-13 in each group were measured by fluorescence quantitative PCR experiment and Western blot experiment, respectively, and the invasion and migration abilities of cells were measured by Transwell and scratch experiments.
RESULTS:
The expression of MMP-13 protein in the positive group and the intervention group was significantly lower than that in the blank group, and the positive group was lower than the intervention group (P<0.05);The mean numbers of invasive cells in blank group, negative group, positive group and intervention group were (1 000.01±44.77), (959.25±46.32), (245.04±4.33), (634.06±33.78) cells/field, respectively;the scratch healing rate of the positive group and the intervention group was significantly lower than that of the blank group, and the positive group was lower than the intervention group (P<0.05).
CONCLUSION
MMP-13 is a target of miR-143, which can reduce the migration and invasion ability of osteosarcoma cells by inhibiting the expression of MMP-13.
Osteosarcoma/pathology*
;
MicroRNAs/genetics*
;
Matrix Metalloproteinase 13/genetics*
;
Neoplasm Invasiveness
;
Animals
;
Mice
;
Cell Line, Tumor
;
Cell Movement
10.Effects of three-dimensional bioprinting antibacterial hydrogel on full-thickness skin defect wounds in rats.
Rong Hua JIN ; Zhen Zhen ZHANG ; Peng Qin XU ; Si Zhan XIA ; Ting Ting WENG ; Zhi Kang ZHU ; Xin Gang WANG ; Chuan Gang YOU ; Chun Mao HAN
Chinese Journal of Burns 2023;39(2):165-174
Objective: To explore the effects of three-dimensional (3D) bioprinting gelatin methacrylamide (GelMA) hydrogel loaded with nano silver on full-thickness skin defect wounds in rats. Methods: The experimental research method was adopted. The morphology, particle diameter, and distribution of silver nanoparticles in nano silver solution with different mass concentrations and the pore structure of silver-containing GelMA hydrogel with different final mass fractions of GelMA were observed by scanning electron microscope and the pore size was calculated. On treatment day 1, 3, 7, and 14, the concentration of nano silver released from the hydrogel containing GelMA with final mass fraction of 15% and nano silver with final mass concentration of 10 mg/L was detected by mass spectrometer. At 24 h of culture, the diameters of inhibition zone of GelMA hydrogel containing final mass concentration of 0 (no nano silver), 25, 50, and 100 mg/L nano silver against Staphylococcus aureus and Escherichia coli were detected. Fibroblasts (Fbs) and adipose stem cells (ASCs) were isolated respectively by enzymatic digestion using the discarded prepuce after circumcision from a 5-year-old healthy boy who was treated in the Department of Urology of the Second Affiliated Hospital of Zhejiang University School of Medicine in July 2020, and the discarded fat tissue after liposuction from a 23-year-old healthy woman who was treated in the Department of Plastic Surgery of the Hospital in July 2020. The Fbs were divided into blank control group (culture medium only), 2 mg/L nano sliver group, 5 mg/L nano sliver group, 10 mg/L nano sliver group, 25 mg/L nano sliver group, and 50 mg/L nano sliver group, which were added with the corresponding final mass concentrations of nano sliver solution, respectively. At 48 h of culture, the Fb proliferation viability was detected by cell counting kit 8 method. The Fbs were divided into 0 mg/L silver-containing GelMA hydrogel group, 10 mg/L silver-containing GelMA hydrogel group, 50 mg/L silver-containing GelMA hydrogel group, and 100 mg/L silver-containing GelMA hydrogel group and then were correspondingly treated. On culture day 1, 3, and 7, the Fb proliferation viability was detected as before. The ASCs were mixed into GelMA hydrogel and divided into 3D bioprinting group and non-printing group. On culture day 1, 3, and 7, the ASC proliferation viability was detected as before and cell growth was observed by live/dead cell fluorescence staining. The sample numbers in the above experiments were all 3. Four full-thickness skin defect wounds were produced on the back of 18 male Sprague-Dawley rats aged 4 to 6 weeks. The wounds were divided into hydrogel alone group, hydrogel/nano sliver group, hydrogel scaffold/nano sliver group, and hydrogel scaffold/nano sliver/ASC group, and transplanted with the corresponding scaffolds, respectively. On post injury day (PID) 4, 7, 14, and 21, the wound healing was observed and the wound healing rate was calculated (n=6). On PID 7 and 14, histopathological changes of wounds were observed by hematoxylin eosin staining (n=6). On PID 21, collagen deposition of wounds was observed by Masson staining (n=3). Data were statistically analyzed with one-way analysis of variance, analysis of variance for repeated measurement, Bonferroni correction, and independent sample t test. Results: The sliver nano particles in nano silver solution with different mass concentrations were all round, in scattered distribution and uniform in size. The silver-containing GelMA hydrogels with different final mass fractions of GelMA all showed pore structures of different sizes and interconnections. The pore size of silver-containing GelMA hydrogel with 10% final mass fraction was significantly larger than that of silver-containing GelMA hydrogels with 15% and 20% final mass fractions (with P values both below 0.05). On treatment day 1, 3, and 7, the concentration of nano silver released from silver-containing GelMA hydrogel in vitro showed a relatively flat trend. On treatment day 14, the concentration of released nano silver in vitro increased rapidly. At 24 h of culture, the diameters of inhibition zone of GelMA hydrogel containing 0, 25, 50, and 100 mg/L nano silver against Staphylococcus aureus and Escherichia coli were 0, 0, 0.7, and 2.1 mm and 0, 1.4, 3.2, and 3.3 mm, respectively. At 48 h of culture, the proliferation activity of Fbs in 2 mg/L nano silver group and 5 mg/L nano silver group was both significantly higher than that in blank control group (P<0.05), and the proliferation activity of Fbs in 10 mg/L nano silver group, 25 mg/L nano silver group, and 50 mg/L nano silver group was all significantly lower than that in blank control group (P<0.05). Compared with the that of Fbs in 0 mg/L silver-containing GelMA hydrogel group, the proliferation activity of Fbs in 50 mg/L silver-containing GelMA hydrogel group and 100 mg/L silver-containing GelMA hydrogel group was all significantly decreased on culture day 1 (P<0.05); the proliferation activity of Fbs in 50 mg/L silver-containing GelMA hydrogel group was significantly increased (P<0.05), while the proliferation activity of Fbs in 100 mg/L silver-containing GelMA hydrogel group was significantly decreased on culture day 3 (P<0.05); the proliferation activity of Fbs in 100 mg/L silver-containing GelMA hydrogel group was significantly decreased on culture day 7 (P<0.05). The proliferation activity of ASCs in 3D bioprinting group show no statistically significant differences to that in non-printing group on culture day 1 (P>0.05). The proliferation activity of ASCs in 3D bioprinting group was significantly higher than that in non-printing group on culture day 3 and 7 (with t values of 21.50 and 12.95, respectively, P<0.05). On culture day 1, the number of dead ASCs in 3D bioprinting group was slightly more than that in non-printing group. On culture day 3 and 5, the majority of ASCs in 3D bioprinting group and non-printing group were living cells. On PID 4, the wounds of rats in hydrogel alone group and hydrogel/nano sliver group had more exudation, and the wounds of rats in hydrogel scaffold/nano sliver group and hydrogel scaffold/nano sliver/ASC group were dry without obvious signs of infection. On PID 7, there was still a small amount of exudation on the wounds of rats in hydrogel alone group and hydrogel/nano sliver group, while the wounds of rats in hydrogel scaffold/nano sliver group and hydrogel scaffold/nano sliver/ASC group were dry and scabbed. On PID 14, the hydrogels on the wound surface of rats in the four groups all fell off. On PID 21, a small area of wounds remained unhealed in hydrogel alone group. On PID 4 and 7, the wound healing rates of rats in hydrogel scaffold/nano sliver/ASC group were significantly higher than those of the other three groups (P<0.05). On PID 14, the wound healing rate of rats in hydrogel scaffold/nano sliver/ASC group was significantly higher than the wound healing rates in hydrogel alone group and hydrogel/nano sliver group (all P<0.05). On PID 21, the wound healing rate of rats in hydrogel alone group was significantly lower than that in hydrogel scaffold/nano sliver/ASC group (P<0.05). On PID 7, the hydrogels on the wound surface of rats in the four groups remained in place; on PID 14, the hydrogel in hydrogel alone group was separated from the wounds of rats, while some hydrogels still existed in the new tissue of the wounds of rats in the other three groups. On PID 21, the collagen arrangement in the wounds of rats in hydrogel alone group was out of order, while the collagen arrangement in the wounds of rats in hydrogel/nano sliver group, and hydrogel scaffold/nano sliver/ASC group was relatively orderly. Conclusions: Silver-containing GelMA hydrogel has good biocompatibility and antibacterial properties. Its three-dimensional bioprinted double-layer structure can better integrate with new formed tissue in the full-thickness skin defect wounds in rats and promote wound healing.
Male
;
Rats
;
Animals
;
Humans
;
Hydrogels/pharmacology*
;
Bioprinting
;
Metal Nanoparticles
;
Rats, Sprague-Dawley
;
Silver/pharmacology*
;
Soft Tissue Injuries
;
Anti-Bacterial Agents

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