1.Mechanism of Modified Si Junzitang and Shashen Maidong Tang in Improving Sensitivity of Cisplatin in EGFR-TKI Resistant Lung Adenocarcinoma Cells Based on Aerobic Glycolysis
Yanping WEN ; Yi JIANG ; Liping SHEN ; Haiwei XIAO ; Xiaofeng YANG ; Surui YUAN ; Lingshuang LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):39-46
ObjectiveTo investigate the mechanism of modified Si Junzitang and Shashen Maidong Tang [Yiqi Yangyin Jiedu prescription (YQYYJD)] in enhancing the sensitivity of cisplatin in epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI)-resistant lung adenocarcinoma cells based on aerobic glycolysis. MethodsThe effects of different concentrations of YQYYJD (0, 2, 3, 4, 5, 6, 7, 8 g·L-1) and cisplatin (0, 3, 6, 9, 12, 15, 18, 21, 24, 27 mg·L-1) on the proliferation and activity of PC9/GR cells were detected by the cell counting kit-8 (CCK-8) assay after 24 hours of intervention. The half-maximal inhibitory concentration (IC50) for PC9/GR cells was calculated to determine the concentrations used in subsequent experiments. PC9/GR cells were divided into blank group (complete medium), YQYYJD group (5 g·L-1), cisplatin group (12 mg·L-1), and combined group (YQYYJD 5 g·L-1 + cisplatin 12 mg·L-1). After 24 hours of intervention, cell viability was measured using CCK-8 assay. Cell proliferation was assessed by colony formation assay, and cell migration was evaluated by scratch and Transwell assays. Glucose consumption, lactate production, and adenosine triphosphate (ATP) levels were measured by colorimetric assays. The expression levels of glycolysis-related proteins, including hexokinase 2 (HK2), phosphofructokinase P (PFKP), pyruvate kinase M2 (PKM2), lactate dehydrogenase A (LDHA), glucose transporter 1 (GLUT1), and monocarboxylate transporter 4 (MCT4), were determined by Western blot. ResultsBoth YQYYJD and cisplatin inhibited the viability of PC9/GR cells in a concentration-dependent manner. The IC50 of PC9/GR cells for YQYYJD and cisplatin were 5.15 g·L-1 and 12.91 mg·L-1, respectively. In terms of cell proliferation, compared with the blank group, the cell survival rate and the number of colonies formed in the YQYYJD group, cisplatin group, and combined group were significantly decreased (P<0.01). Compared with the YQYYJD and cisplatin groups, the combined group showed a further significant reduction in cell survival rate and colony formation (P<0.01). In terms of cell migration, compared with the blank group, the cell migration rate and the number of cells passing through the Transwell membrane in the YQYYJD group, cisplatin group, and combined group were significantly decreased (P<0.01). Compared with the YQYYJD and cisplatin groups, the combined group exhibited a further significant reduction in cell migration rate and the number of cells passing through the Transwell membrane (P<0.01). In terms of glycolysis, compared with the blank group, glucose consumption, lactate production, and ATP levels in the YQYYJD group, cisplatin group, and combined group were significantly decreased (P<0.01). Compared with the YQYYJD and cisplatin groups, the combined group showed a further significant reduction in glucose consumption, lactate production, and ATP levels (P<0.05). Compared with the blank group, the protein expression levels of HK2, PFKP, PKM2, and LDHA in the YQYYJD, cisplatin, and combined groups were significantly decreased (P<0.01). The combined group showed a further significant reduction in the expression levels of these proteins compared with the YQYYJD and cisplatin groups (P<0.01). No significant differences were observed in the protein expression levels of GLUT1 and MCT4 among the groups. ConclusionYQYYJD can synergistically inhibit the proliferation and migration of PC9/GR cells and enhance their sensitivity to cisplatin. The mechanism may be related to the downregulation of the expression of glycolysis-related rate-limiting enzymes, including HK2, PFKP, PKM2, and LDHA, thereby inhibiting glycolysis.
2.Impact factor selection for non-fatal occupational injuries among manufacturing workers by LASSO regression
Yingheng XIAO ; Chunhua LU ; Juan QIAN ; Ying CHEN ; Yishuo GU ; Zeyun YANG ; Daozheng DING ; Liping LI ; Xiaojun ZHU
Journal of Environmental and Occupational Medicine 2025;42(2):133-139
Background As a pillar industry in China, the manufacturing sector has a high incidence of non-fatal occupational injuries. The factors influencing non-fatal occupational injuries in this industry are closely related at various levels, including individual, equipment, environment, and management, making the analysis of these influencing factors complex. Objective To identify influencing factors of non-fatal occupational injuries among manufacturing workers, providing a basis for targeted interventions and surveillance. Methods A total of
3.Construction of a system for isolation and purification of NK cells from whole blood donations
Tengyu CAO ; Huayu LIN ; Xuanzhi ZHANG ; Cuimi DUAN ; Yi LIU ; Xiaonan XUE ; Liping SUN ; Yang YU
Chinese Journal of Blood Transfusion 2025;38(2):181-188
[Objective] To explore the feasibility of using whole blood as a source of NK cells for allogeneic CAR NK cell therapy and activated NK cell reinfusion therapy, and initially construct a technical system for the separation and purification of NK cells from whole blood. [Methods] All peripheral blood mononuclear cells (PBMCs) were enriched from 400 mL of whole blood by manual separation and machine separation, respectively. The erythrocyte loss rate, PBMCs number, NK cell purity of the two methods were compared. NK cells were sorted from PBMCs by three separation and enrichment methods as immunomagnetic bead negative selection method, platelet lysate culture expansion and PERCOLL density gradient separation method, and the purity and yield of NK cells, the activity of NK cells and the tumor-killing ability of the three separation and enrichment methods were compared. [Results] The proportion of NK cells in the lymphocyte population was higher in the manual separation method than in the machine separation method[(13.16±5.16)% vs (8.56±3.92)%, P<0.05]; the number PBMCs was lower in the manual separation method than in the machine separation method[(4.09±1.80)×108vs (6.49±2.16)×108, P<0.05], and there was no difference in the red blood cell loss between the two methods (P>0.05). The purity of NK cells isolated and enriched from PBMCs by manual separation method using immunomagnetic was (96.77±2.31)%; the yield was (56.27±10.47)%; the inhibition of tumor proliferation was (38.67±14.05)%; and the tumor killing rate was (19.90±8.05)%. The purity of NK cells isolated and enriched from PBMCs by manual separation method using platelet lysis culture expansion method was the highest at day 7, which was (54.84±15.80)%; the cell expansion multiple could reach 16.92±6.28 at day 7; the in vitro tumor killing rate of NK cells was (15.83±5.5)%; the tumor inhibition rate was (44.33±13.5)%; and there was no difference in the toxicity and activity of NK cells between the two methods (P>0.05). The purity of NK cells isolated and enriched by PERCOLL density gradient separation method was (15.83±5.82)%, and the yield was (14±6.25)%, which was significantly lower than the other two methods. [Conclusion] PBMCs isolated from whole blood by manual separation and NK cells enriched by negative selection with immunomagnetic beads have the potential to provide NK cell materials for CAR-NK cell therapy, and NK cells enriched by platelet lysate-conditioned medium have the potential to provide NK cells for large-scale NK cell activation reinfusion therapy.
4.Degradation dynamics and residue determination of pyriclobenzuron in rice and paddy environments
Huanqi WU ; Junmin WANG ; Kongtan YANG ; Xumi WANG ; Nan FANG ; Liping DUAN ; Changpeng ZHANG ; Xiangyun WANG
Chinese Journal of Schistosomiasis Control 2025;37(2):201-208
Objective To establish a method for determination of pyriclobenzuron (PBU) residues in rice and paddy environments, and to determine the residual amounts and observe the degradation dynamics of PBU. Methods In July 2022, the paddies of Zhejiang Academy of Agricultural Sciences were selected as experimental fields, and were divided into the blank control group (no pesticide application), the 1-fold-concentration pesticide group (1 kg/667 m2), and the 5-fold-concentration pesticide group (5 kg/667 m2), with a 100 m2 area in each group. At the early tillering stage of rice, 20% suspension of PBU sulfate was sprayed once in the 1-fold-concentration and 5-fold-concentration pesticide groups, and rice plants, paddy water and soil samples were collected 2 h, and 1, 2, 3, 5, 7, 11, 14, 21, 28, 35, 49 d and 63 d following spraying PBU, while rice straw, field soil, brown rice and rice husk samples were collected 98 d following spraying. PBU was extracted and purified in samples using a quick, easy, cheap, effective, rugged, and safe (QuEChERS) pretreatment technique, and the PBU contents were determined in samples using ultrahigh performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The solvent standard working solution and matrix standard working solution were prepared. A linear regression equation was fitted between PBU concentration (x-axis) and peak area (y-axis), and the ratio of the slope (k) of the matrix standard curve to the slope (K) of the solvent standard curve was calculated to evaluate the matrix effect of PBU in samples. According to the Guidelines for Pesticide Residue Testing in Crops (NY/T 788—2018), the addition levels of PBU were set at 0.005, 0.050, 5.000, 1 000.000 mg/kg in rice plants, 0.005, 0.050, 2.000, 10.000 mg/kg in paddy water, 0.005, 0.050, 2.000 mg/kg in soil, and 0.005, 0.050, 5.000 mg/kg in brown rice and rice husks. The recovery and relative standard deviation (RSD) of PBU addition were calculated to evaluate the effectiveness of UPLC-MS/MS for determination of PBU contents. The first-order kinetic equation of PBU concentration was fitted in samples at different sampling time points to analyze the trends in PBU degradation in rice plants, paddy water, and soil, and the half-life of PBU was calculated in different samples. Results There was a good linear relationship between the mass concentration and peak area of PBU at concentrations of 0.000 1 to 0.020 0 mg/kg under solvent and matrix conditions (R2 = 0.985 8 to 0.999 7, t = -0.47 to 1.62, all P values < 0.01). The matrix effects of PBU were 70.26%, 65.42% and 65.12% in rice plants, brown rice and rice husks, indicating a matrix-inhibitory effect, and the matrix effect was 87.06% in soils, indicating a weak matrix effect. The recovery of PBU addition was 77.61% to 100.12% in different samples, with RSD of 1.43% to 6.74%, and a limit of quantification (LOQ) of 0.005 mg/kg, and the addition recovery and RSD met the requirements of the Guidelines for Pesticide Residue Testing in Crops (NY/T 788—2018), validating the effectiveness of UPLC-MS/MS assay. Following spraying PBU at a dose of 1 kg/667 m2, the half-life of PBU was 6.24 d in rice plants and 3.43 d in paddy water samples, respectively. The final residues of PBU were lower than the LOQ of 0.005 mg/kg in brown rice and rice husk samples 98 d following spraying PBU. Following spraying PBU at a dose of 5 kg/667 m2, the half-life of PBU was 15.75 d in rice plants and 7.62 d in paddy water samples, respectively. The final residue of PBU was lower than the LOQ of 0.005 mg/kg in brown rice 98 d following spraying PBU, and the final residue of PBU was 0.049 mg/kg in rice husks. Conclusions A simple, and highly accurate and precise UPLC-MS/MS assay has been developed for determination of PBU residues in rice plants and paddy environments through extraction and purification of PBU from matrix samples using QuEChERS pretreatment. After spraying PBU in paddies, the concentration of PBU gradually decreases in rice plants and paddy water over time, and the final residual concentration is low.
5.Herbal Textual Research on Kochiae Fructus in Famous Classical Formulas
Huifang HU ; Liping YANG ; Fei CHEN ; Xiaohui MA ; Ling JIN ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(15):247-257
In this paper, by referring to ancient and modern literature, the textual research of Kochiae Fructus has been conducted to clarify the name, origin, distribution of production areas, quality specification, taste and efficacy, harvesting time, processing and compatibility taboo, so as to provide reference and basis for the development and utilization of related famous classical formulas. According to the investigation, it can be seen that Difuzi was first published in Sheng Nong's Herbal Classic, and has been used as the official name throughout history. It is also known by other names such as Dimai, Dikui, and Luozhou. The mainstream source of Difuzi in materia medica throughout history is the dried ripe fruit of Kochia scoparia, which is consistent throughout history. In the Han dynasty, it was recorded that Kochiae Fructus was produced in Jingzhou(Hubei province), while modern literature records its distribution throughout the country, so it does not have obvious geoherbalism. The harvesting period of Kochiae Fructus is mostly in the late autumn, and the quality is best when it is full, gray green in color, and no impurities. There are two processing methods for its origin:from the Southern and Northern dynasties to the Ming dynasty, it was dried in the shade, and after the founding of the People's Republic of China, it was dried in the sun. There are few records about the processing of Kochiae Fructus, and its clinical application is mostly based on raw products as medicine. The seedlings are harvested in February of the lunar calendar, and the leaves are taken in April and May, processing in the place of origin is shade drying, the processing methods include burning ash and frying frost, pounding juice and wine soaking. For internal use, it is mostly decocted or mashed, while for external use, it is mostly washed with decoction or taken in a soup bath. Throughout history, it has been recorded that Kochiae Fructus is bitter and cold, and is mainly used for treating bladder fever. After the founding of the People's Republic of China, most of the literature classified it as damp-clearing medicine. Since the 1985 edition of Chinese Pharmacopoeia, it has been recorded that Kochiae Fructus has a pungent and bitter taste, and a cold nature. Returning to the kidney and bladder meridians with functions of clearing heat and dampness, dispelling wind and relieving itching. The clinical contraindications are mainly prohibited for those with deficiency and no dampness and heat. Throughout history, it has been recorded that the taste of the seedlings and leaves is bitter and cold for treatment of dysentery. Since modern times, it has been used to regulate the liver, spleen and large intestine meridians, with functions such as clearing heat and detoxifying, and diuresis. Based on the textual research, it is recommended to use the dried ripe fruit of K. scoparia when developing the famous classical formulas containing Kochiae Fructus, and processing shall be carried out according to the original processing requirements. If the original formula does not specify the processing requirements, the raw products is taken into medicine.
6.Differential Analysis on Chemical Composition and Pharmacodynamic Effect Between Combined Decoction and Single Decoction of Famous Classical Formula Huaganjian
Yang WANG ; Gaoju ZHANG ; Ling LI ; Liping CHEN ; Li ZHANG ; Xiao LIU ; Yuyu ZHANG ; Yuan CUI ; Minglong LI ; Chaomei FU ; Xin YAN ; Yuxin HE ; Qin DONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):199-207
ObjectiveThrough qualitatively and quantitatively analysis of the differences in chemical composition between the combined decoction and single decoction of Huaganjian and comparison of their core efficacy, to explore the rationality of the flexible clinical application of Huaganjian compound preparations and single-flavored dispensing granules. MethodsUltra performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry(UPLC-Q-Exactive Orbitrap MS) was used to qualitatively analyze the combined decoction and single decoction samples of Huaganjian, and meanwhile, the contents of four index components(geniposide, paeoniflorin, hesperidin and paeonol) were quantitatively analyzed by high performance liquid chromatography(HPLC). Nonalcoholic fatty liver disease(NAFLD) rat model induced by high-fat diet was applied to compare the efficacy of combined decoction and single decoction of Huaganjian. A total of 30 male SD rats were randomly divided into the control group, model group, lovastatin group(1.8 mg·kg-1), combined decoction group(1.26 g·kg-1) and single decoction group(1.18 g·kg-1). After successful modeling, lovastatin group, combined decoction group and single decoction group were given corresponding doses of drugs by intragastric administration every day, and the control group and model group were given equal amounts of normal saline by intragastric administration, after 4 weeks of administration, the serum and liver tissues were collected, and the contents of alanine aminotransferase(ALT), aspartate aminotransferase(AST), total cholesterol(TC), triglyceride(TG), low-density lipoprotein cholesterol(LDL-C) and high-density lipoprotein cholesterol(HDL-C) in serum of rats were detected, and the liver pathological examination was carried out by hematoxylin-eosin(HE) staining and oil red O staining, so as to compare differences of their efficacy. ResultsSeventy chemical components were initially identified and attributed from the lyophilized powder of the combined decoction and single decoction samples of Huaganjian, and there was no obvious difference in composition between the two. Further quantitative analysis showed that the contents of geniposide, paeoniflorin, hesperidin and paeonol in the combined decoction samples were significantly increased when compared with those of the single decoction samples(P<0.01). The pharmacodynamic results showed that compared with the model group, both the combined and single decoction groups of Huaganjian could improve the liver index of NAFLD rats, reduce the serum levels of AST, ALT, TC, TG and LDL-C, increase the serum level of HDL-C, and ameliorate the pathological changes of liver cell steatosis and fat accumulation. However, there was no significant difference in pharmacodynamic effects between the combined decoction group and the single decoction group. ConclusionThere is no significant difference between the combined decoction and single decoction of Huaganjian in terms of chemical composition, but the contents of the four index components show significantly difference. Both of them can significantly improve the fat accumulation and liver function in NAFLD rats. This study provides a reference basis for the rational clinical application and evaluation of famous classical formula compound preparations and single-flavored dispensing granules.
7.Herbal Textual Research on Cnidii Fructus in Famous Classical Formulas
Huifang HU ; Liping YANG ; Fei CHEN ; Xiaohui MA ; Ling JIN ; Zhilai ZHAN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(16):243-253
In this paper, by referring to ancient and modern literature, the textual research of Cnidii Fructus has been conducted to clarify the name, origin, distribution of production areas, quality specification, nature and flavour, efficacy, harvesting and processing, compatibility taboo and others, so as to provide reference and basis for the development and utilization of the relevant famous classical formulas. After textual research, it can be verified that Cnidii Fructus was first published in Sheng Nong's Herbal Classic, the materia medica of all dynasties was named Shechuangzi, and there are also aliases such as Shesu, Shemi, and Qiangmi. The main source for generations was the dried ripe fruit of Cnidium monnieri, and ancient and modern consistent. From the Eastern Han dynasty to Tang dynasty, the origin of Cnidii Fructus was Zibo, Shandong province. During the Five dynasties, it expanded to Yangzhou in Jiangsu province and Xiangyang in Hubei province, the Song dynasty added Shangqiu in Henan province, and it was considered that Yangzhou, Xiangyang and Shangqiu were its genuine producing areas. It was more widely distributed in Ming and Qing dynasties. After the founding of the People's Republic of China, the origin is clearly distributed throughout the country. For its quality evaluation, generally full grain, gray yellow color, strong aroma is the best. The harvesting period in the past dynasties was mostly the fifth lunar month, and the fruit was collected to remove impurities and dry. The mainstream processing in producing area of the past dynasties was net selection of raw products, mixing and steaming with the juice of Rehmanniae Radix and stir-frying were the mainstream processing methods in the past, there were also stir-frying with honey, stir-frying with salt and rice wine, immersing and steaming with rice wine and other methods. In recent times, it has been used in raw products as medicine. Sheng Nong's Herbal Classic recorded Cnidii Fructus was bitter, Supplementary Records of Famous Physicians recorded its acrid for the first time. It was recorded in the Ming dynasty that its nature was warm, acted on the kidney meridian, and had small toxicity. After the founding of the People's Republic of China, most of the literature classified it as a medicine to attack poison, kill insects and relieve itching with the functions of dispelling pathogenic wind and removing dampness, destroying parasites and elieving itching, warming kidney and activating Yang. Clinical contraindications are mainly contraindicated for people with damp-heat from the lower-jiao or kidney heat. Based on the textual research, it is suggested that when developing the famous classical formulas containing Cnidii Fructus, the source shall be the dried ripe fruit of C. monnieri, and then it shall be processed according to the original formulas. If there is no requirement for processing in the formulas, the raw products can be taken into medicine.
8.Mechanism of Kaixuan Jiedu Core Prescription in Regulating PTGS2 to Improve Skin Lesions in Psoriasis Mouse Models
Xue XIAO ; Liping KANG ; Dan DAI ; Yidi MA ; Bin YANG ; Ping SONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):49-59
ObjectiveTo identify the active constituents of Kaixuan Jiedu core prescription (KXJD) and investigate its effective components and therapeutic targets in the treatment of common psoriasis
9.Mechanism of Kaixuan Jiedu Core Prescription in Regulating PTGS2 to Improve Skin Lesions in Psoriasis Mouse Models
Xue XIAO ; Liping KANG ; Dan DAI ; Yidi MA ; Bin YANG ; Ping SONG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(17):49-59
ObjectiveTo identify the active constituents of Kaixuan Jiedu core prescription (KXJD) and investigate its effective components and therapeutic targets in the treatment of common psoriasis
10.Construction and evaluation of a multi-base collaborative training system for anticoagulation specialty clinical pharmacists
Shujie DONG ; Liping DU ; Yatong ZHANG ; Zheng DING ; Wenxing PENG ; Zinan ZHAO ; Xiaoxiao LI ; Li YANG
China Pharmacy 2025;36(15):1837-1840
OBJECTIVE To enhance the training quality of anticoagulation specialty clinical pharmacists, address the resource limitations of a single training base, and promote homogenization of training quality. METHODS A multi-base joint training system for anticoagulation specialty clinical pharmacists in the Beijing area was established. A mixed research method was employed, collecting data through performance comparisons, questionnaires, and qualitative interviews to compare the differences between the joint training model (experimental group, n=16) and traditional teaching model (the control group, n=17). RESULTS The established joint training system encompassed a unified joint training teaching plan, the formation of a joint training teaching team, the establishment of joint theoretical teaching courses, the implementation of joint case discussions and literature presentations, as well as strengthening the assessment throughout the joint training process. Compared to the control group [theoretical assessment of (76.44±3.66) points, case assessment of (84.31±3.27) points], the experimental group students achieved higher scores in theoretical assessment ([ 79.85±4.64) points] and case assessment ([ 88.70±5.51) points] (P<0.05). Through questionnaires and qualitative interviews, the trainees in experimental group were highly satisfied with the joint training model in terms of theoretical learning, communication skills, and teaching interaction. CONCLUSIONS The multi-base collaborative training system for anticoagulation specialty clinical pharmacists can integrate advantageous resources and significantly enhance the training effectiveness of anticoagulation specialty clinical pharmacists, offering value for wider promotion.

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