ObjectiveTo observe the effect of M1 bone marrow-derived macrophages （M1-BMDM） with Wnt5a knockdown on liver fibrosis and regeneration in a rat model of liver cirrhosis， and to investigate its gain-of-function effect compared with unmodified M1-BMDM. MethodsPrimary bone marrow-derived macrophages were isolated from rats and were polarized to M1 phenotype to construct M1-BMDM^Wnt5a-KD cells. A rat model of liver cirrhosis induced by CCl₄/2-AAF was established， and at the end of week 8， rats were randomly divided into model group， M1-BMDM group， M1-BMDM Wnt5a-knockdown empty vector group （M1-BMDM^KD-EV group）， and M1-BMDM Wnt5a-knockdown group （M1-BMDM^Wnt5a-KD group）， with 6 rats in each group. On the first day of week 9， the rats in each group were given a single injection of the corresponding cells via the caudal vein， along with an intraperitoneal injection of a CCR2 inhibitor. Six rats without any treatment were used as normal control group. Samples were collected at the end of week 12 to assess liver histopathology， serum liver function parameters， hepatic stellate cell activation， and the expression levels of mature hepatocyte markers. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the model group， all cell treatment groups had significant alleviation of liver inflammatory response and significant reductions in the activities of alanine aminotransferase and aspartate aminotransferase （AST） in serum （all P<0.01）， and the M1-BMDM^Wnt5a-KD group had a significantly lower serum level of AST than the M1-BMDM group （P<0.05）. The semi-quantitative analysis based on immunohistochemical staining showed that compared with the model group， all cell treatment groups had a significant reduction in the percentage of CD68-positive area （all P<0.05）， and compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had a significant reduction in the percentage of CD68-positive area and a significant increase in the percentage of CD163-positive area （both P<0.05）. Compared with the model group， all cell treatment groups had significant reductions in the mRNA expression levels of CD68 and tumor necrosis factor-α （all P<0.05） and the protein expression level of CD68 （all P<0.01）； compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had significant increases in the protein and mRNA expression levels of CD163 （both P<0.05）， significant reductions in the protein and mRNA expression levels of CD68 （both P<0.05）， and a significant reduction in the protein expression level of tumor necrosis factor-α （P<0.01）. Sirius Red collagen staining and alpha-smooth muscle actin （α-SMA） immunohistochemical staining showed that compared with the model group， all cell treatment groups had significant alleviation of liver collagen deposition and α-SMA-positive area， with the most significant changes in the M1-BMDM^Wnt5a-KD group， and compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had significantly smaller Sirius Red-positive area and α-SMA-positive area and a significantly lower content of hydroxyproline in liver tissue （all P<0.05）. Compared with the M1-BMDM^KD-EV group， the M1-BMDM^Wnt5a-KD group had significant reductions in the protein and mRNA expression levels of α-SMA and the mRNA expression level of COL-I and TGF-β （all P<0.05）. Compared with the model group， all cell treatment groups had a significant increase in the protein expression level of HNF-4α in liver tissue （all P<0.05）， and the M1-BMDM^Wnt5a-KD group had significantly higher protein and mRNA expression levels of HNF-4α and hepatocyte specific antigen than the M1-BMDM^KD-EV group （both P<0.05）. The M1-BMDM^Wnt5a-KD group had a significantly higher serum level of albumin than the M1-BMDM^KD-EV group （P<0.01）. Immunofluorescence co-staining showed that compared with the model group， all cell treatment groups had a significant increase in the number of cells stained positive for HNF and HNF-4α and Ki67 （all P<0.01）， and the M1-BMDM^Wnt5a-KD group had a significantly higher number of such cells than the M1-BMDM^KD-EV group （P<0.05）. ConclusionInhibition of Wnt5a expression enhances the therapeutic effect of M1-BMDM on rats with liver cirrhosis induced by CCl₄/2-AAF， which provides new ideas for enhancing the anti-cirrhotic effect of M1-BMDM through genetic modification.

ObjectiveTo investigate the effect and mechanism of transplantation of human umbilical cord mesenchymal stem cell （hUC-MSC） with overexpression of the Numb gene in the treatment of cholestatic liver fibrosis （CLF）. MethodsThe technique of lentiviral transfection was used to induce the overexpression of the Numb gene in hUC-MSC （hUC-MSC^Numb-OE）， and hUC-MSC transfected with empty vector （hUC-MSC^OE-EV） was used as negative control. Bile duct ligation （BDL） was performed to establish a rat model of CLF， and then the rats were randomly divided into BDL group， hUC-MSC group， hUC-MSC^OE-EV group， and hUC-MSC^Numb-OE group， while a sham-operation group was also established. The rats in the intervention groups were given a single splenic injection of the corresponding cells after BDL， and samples were collected at the end of week 4. Related indicators were measured， including serum biochemistry， liver histopathology， the content of hydroxyproline （Hyp） in the liver， hepatic stellate cell activation， ductular reaction， liver regeneration， and the expression levels of key molecules in the Numb-p53 signaling axis. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the BDL group， the hUC-MSC group and the hUC-MSC^OE-EV group had significant reductions in the levels of serum biochemical parameters （aspartate aminotransferase， gamma-glutamyl transpeptidase， total bile acid， total bilirubin， and direct bilirubin）， liver fibrosis markers （the content of Hyp and the expression levels of alpha-smooth muscle actin， tumor necrosis factor-α， and transforming growth factor-beta 1）， and ductular reaction markers （the expression levels of CK7 and CK19） （all P <0.05）， and compared with the hUC-MSC^OE-EV group， the hUC-MSC^Numb-OE group had significantly greater improvements in the above indicators （all P <0.05）. In addition， compared with the hUC-MSC^OE-EV group， the hUC-MSC^Numb-OE group had significant improvements in the expression levels of liver regeneration-related markers （albumin and hepatocyte nuclear factor 4α） and the molecules associated with the Numb-p53 signaling axis （Numb， pNumb， Mdm2， and p53） （all P <0.05）. ConclusionOverexpression of the Numb gene can enhance the therapeutic effect of hUC-MSC on CLF， possibly by activating the Numb-PTB^L-p53-HNF4α axis， promoting the hepatic differentiation of hUC-MSCs and subsequently enhancing liver regeneration.

OBJECTIVES: To reprogram human placental fibroblasts (HPFs) into chemically induced epithelioid-like cells (ciEP-Ls) using a combination of small-molecule compounds. METHODS: HPFs cultured under normoxic conditions were identified using immunofluorescence assay, PCR and chromosomal karyotyping. Under hypoxic conditions (37 ℃, 5% O₂), HPFs were cultured in a medium containing small-molecule compounds C6TSEDRVAJZ (CHIR99021, 616452, TTNPB, SAG, EPZ5676, DZNep, Ruxolitinib, VTP50469, Afuresertib, JNK-IN-8, and EZM0414), and the cell morphology was observed daily. The expression levels of epithelial cell markers in the induced cells were detected by immunofluorescence, Western blotting and PCR. Chromosomal karyotyping of the induced cells was performed and the induction efficiency was calculated. RESULTS: Before induction, HPFs showed positive expressions of fibroblast surface markers CD34 and vimentin and were negative for epithelial surface markers. PCR results showed high expressions of fibroblast-specific genes S100A4 and COL1A1 in HPFs with a normal human diploid karyotype. After one day of induction, the HPFs underwent morphological changes from a multinodular spindle shape to a round or polygonal shape, which was morphologically characteristic of ciEP-Ls. On day 4 of induction, the cells exhibited high expressions of the epithelial cell markers E-cadherin and Lin28A. RT-qPCR results also showed that the cells expressed the epithelial markers Smad3, GLi3, PAX8, WT1, KRT19, and KRT18 with significantly down-regulated expressions of all the fibroblast surface markers and a normal human diploid karyotype. The reprogramming efficiency of HPFs into ciEP-Ls ranged from (64.53±2.8)% to (68.10±3.6)%. CONCLUSIONS The small-molecule compound combination C6TSEDRVAJZ is capable of inducing HPFs into ciEP-Ls under hypoxic conditions with a high induction efficiency.
Humans ; Fibroblasts/drug effects* ; Pregnancy ; Female ; Cell Differentiation/drug effects* ; Pyrimidines/pharmacology* ; Placenta/cytology* ; Cells, Cultured ; Pyridines/pharmacology* ; Pyrazoles/pharmacology* ; Epithelial Cells/cytology*

OBJECTIVES: To explore the efficacy of DSA-guided intrathecal drug delivery system combined with Zi Wu Liu Zhu Acupoint Therapy for management of cancer pain and provide reference for its standardized clinical application. Methods and. RESULTS: Recommendations were formulated based on literature review and expert group discussion, and consensus was reached following expert consultation. The consensus recommendations are comprehensive, covering the entire treatment procedures from preoperative assessment and preparation, surgical operation process, postoperative management and traditional Chinese medicine treatment to individualized treatment planning. The study results showed that the treatment plans combining traditional Chinese with Western medicine effectively alleviated cancer pain, reduced the use of opioid drugs, and significantly improved the quality of life and enhanced immune function of the patients. Postoperative follow-up suggested good treatment tolerance among the patients without serious complications. CONCLUSIONS The formulated consensus is comprehensive and can provide reference for clinicians to use DSA-guided intrathecal drug delivery system combined with Zi Wu Liu Zhu Acupoint Therapy. The combined treatment has a high clinical value with a good safety profile for management of cancer pain.
Humans ; Medicine, Chinese Traditional ; Cancer Pain/therapy* ; Drugs, Chinese Herbal/therapeutic use* ; Drug Delivery Systems ; Pain Management/methods* ; China

Objective:To explore the therapeutic effect of micro-flaps carrying sensory nerve in treatment of high-pressure injection injuries of the digit (HPIID).Methods:From January 2022 to June 2024, retrospective analysis of 7 patients who had HPIID were admitted to the Department of Hand Surgery Division 1, Norinco General Hospital. The patients were 5 males and 2 females with ages from 25 to 59 years. The digital injuries were: 3 index fingers, 2 middle fingers and 2 thumbs. All patients received debridement under microscope in primary surgery, with the defects at 1.5 cm×2.4 cm - 2.5 cm×5.5 cm in size after debridement. In stage Ⅱ surgery, 5 patients received the treatment of free fibular great toe flap carrying peroneal nerve of the great toe, with the flap size at 1.8 cm× 2.0 cm - 2.7 cm×4.0 cm. Two patients received the treatment of transfer of free fibular medial plantar flap carrying medial plantar nerve, with flap size at 1.9 cm×2.6 cm - 4.5 cm×5.7 cm. Donor sites were directly sutured in 5 patients, and 2 patients received skin grafting. Five patients received postoperative follow-up at outpatient clinic, 1 patient via telephone interview and 1 via WeChat review.Results:All 7 flaps survived and all wounds had primary healing. All donor and recipient sites and skin grafting sites healed primarily. Postoperative follow-up lasted for 4 to 9 months, with an average of 6.4 months. All the affected digits had satisfactory appearance and function, except 1 which was slightly slimmer than the healthy side. Range of motion of the affected digits was evaluated according to total active movement (TAM): 5 were in excellent and 2 in good. Sensory recovery of the digits was evaluated according to the British Medical Research Council (BMRC): 1 digit was at S 2, 2 at S 3 and 4 at S 4. One patient had two-point discrimination (TPD) at 9.0-15.0 mm, 2 at 6.0-10.0 mm, and 4 at 3.0-6.0 mm. Conclusion:For HPIID with a defect, surgical treatment with transfer of micro-flap carrying sensory nerve should be a preferred treatment option.

Objective To explore the efficacy of DSA-guided intrathecal drug delivery system combined with Zi Wu Liu Zhu Acupoint Therapy for management of cancer pain and provide reference for its standardized clinical application.Methods and Results Recommendations were formulated based on literature review and expert group discussion,and consensus was reached following expert consultation.The consensus recommendations are comprehensive,covering the entire treatment procedures from preoperative assessment and preparation,surgical operation process,postoperative management and traditional Chinese medicine treatment to individualized treatment planning.The study results showed that the treatment plans combining traditional Chinese with Western medicine effectively alleviated cancer pain,reduced the use of opioid drugs,and significantly improved the quality of life and enhanced immune function of the patients.Postoperative follow-up suggested good treatment tolerance among the patients without serious complications.Conclusion The formulated consensus is comprehensive and can provide reference for clinicians to use DSA-guided intrathecal drug delivery system combined with Zi Wu Liu Zhu Acupoint Therapy.The combined treatment has a high clinical value with a good safety profile for management of cancer pain.

[Objective]To explore the academic characteristics and the current status of active inheritance of the acupuncture and moxibustion schools in northern Zhejiang.[Methods]Through literature search,data query,personnel interview and other ways,collect the relevant records of acupuncture and moxibustion in northern Zhejiang,sort out the representative schools and summarize their common academic characteristics.[Results]Upon reviewing the literature,it was discovered that the acupuncture and moxibustion schools in northern Zhejiang had a long and storied history,holding a significant position within the academic system of the"Xiushui medical school".Through historical medical research and interviews,it was summarized that the acupuncture and moxibustion in northern Zhejiang possessed five major academic characteristics:precise acupoint selection,diligent study of classics,skillful application of techniques,compassionate medical practice and emphasis on moxibustion therapy.However,the schools faced challenges such as slow transmission,traditional methods and limited dissemination scope during the process of inheritance.[Conclusion]The acupuncture and moxibustion schools in northern Zhejiang have demonstrated distinct academic characteristics and potential for intergenerational innovation in acupoint theory,manual skills and humanistic care.Their inheritance practices have deepened the academic connotations of the"Xiushui medical school".In the future,it is essential to enhance the current status of active inheritance through modern technology and communication methods to promote the widespread dissemination and sustainable development of the academic ideas of northern Zhejiang acupuncture and moxibustion.

The paper introduces Professor JI Laixi's academic thought and clinical experience in treatment of primary open angle glaucoma with "nape-eight-needle" acupotomy. Professor JI Laixi believes that the key pathogenesis lies in "occlusion of xuanfu (subtle orifices) within the eyes and obstruction of meridian pathways". Using the unblocking principle of treatment, taking meridian theory of traditional acupuncture as the core and based on the anatomical principles of structural acupuncture, Professor JI has proposed his academic thought, "treating eye diseases from the nape". In treatment, "nape-eight-needle" acupotomy is adopted, combined with filiform needle acupuncture. It is the advantageous compound therapeutic method, aiming to open xuanfu, restore brain-eye meridian connectivity, harmonize body, qi and mind through systemic regulation, address both the causative factors and symptoms and prevent from blindness. This therapeutic approach provides a new idea for clinical treatment of primary open angle glaucoma.
Humans ; Acupuncture Therapy/history* ; Glaucoma, Open-Angle/therapy* ; Male ; Meridians ; Female ; Acupuncture Points ; Middle Aged ; Aged

Objective:To investigate the effect and potential mechanism of circular RNA-centrosome and spindle pole-associated protein 1 (circ-CSPP1) on the malignant biological behavior of hepatoma cells.Methods:Real-time fluorescence quantitative polymerase chain reaction (RT-PCR) was used to detect the expressions of circ-CSPP1 and microRNA-582-5p (miR-582-5p) in hepatoma cells, and Western blotting was used to detect the expression of karyopherins α2 (KPNA2). HepG2 cells were divided into the circ-CSPP1 overexpression group, the circ-CSPP1 overexpression control group, the si-CSPP1 group, the si-NC group, the si-CSPP1+ miR-582-5p inhibition group, and the si-CSPP1+ miR-582-5p inhibition control group. circ-CSPP1 overexpression plasmid, CSPP1 interfering small RNA, CSPP1 interfering small RNA, miR-582-5p inhibition sequence and negative control were transfected respectively in these groups. Cell proliferation in each group was detected by 5-acetylene-2'-deoxyuridine (Edu), invasion ability was detected by Transwell assay, and the binding of circ-CSPP1 and KPNA2 to miR-582-5p was verified by dual-luciferase assay. In the si-CSPP1 group, HepG2 cells transfected with si-CSPP1 lentivirus were subcutaneously injected into the back of nude mice ( n=12), and in the si-NC group, HepG2 cells transfected with negative control lentivirus ( n=12) were injected. The tumor mass, volume, circ-CSPP1 and KPNA2 were detected. Results:In the circ-CSPP1 overexpression group, the relative expression of circ-CSPP1 was (1.68±0.17), the expression of KPNA2 was (1.52±0.16), and the number of invasive cells in the 100-fold field of view was (128.4±13.5), which were all higher than those in the circ-CSPP1 overexpression control group [(1.25±0.16), (1.24±0.15), (128.4±13.5)], while the expression of miR-552-5p was lower than that in the circ-CSPP1 overexpression control group [(0.96±0.11) vs (1.31±0.15)]; The relative expression of circ-CSPP1 in the si-CSPP1 group was (1.02±0.13), KPNA2 was (0.74±0.09), and the number of invasive cells was (53.5±6.7), which were lower than those in the si-NC group [(1.28±0.14), (1.22±0.13), (74.6±8.3)], while the expression of miR-582-5p was higher than that in the si-NC group [(1.71±0.18) vs (1.32±0.14)]; The expression of circ-CSPP1 and KPNA2 and the number of invasion cells in the si-CSPP1+ miR-582-5p inhibition group was higher than that in the si-CSPP1+ miR-582-5p inhibition control group, and the differences were statistically significant (all P<0.05). The results of cell proliferation were consistent with those of invasion. The dual-luciferase gene report showed that, compared with the miR-NC group, the relative luciferase activity in HepG2 cells co-transfected with circ-CSPP1-WT or KPNA2-WT wild-type reporter vectors in the miR-882-5p mimic group decreased [(0.46±0.05) vs (1.03±0.11), (0.42±0.03) vs (1.01±0.09)]. The differences were all statistically significant (both P<0.05). However, there was no statistically significant difference in the relative luciferase activity in HepG2 cells co-transfected with the circ-CSPP1-MUT or KPNA2-MUT mutant reporter vectors (both P>0.05). The tumor weight, volume and circ-CSPP1 and KPNA2 expressions in tumor tissue of nude mice in the si-CSPP1 group were all lower than those in the si-NC group, and the expression of miR-582-5p was higher than that in the si-NC group. The differences were statistically significant (all P<0.05). Conclusion:Inhibition of circ-CSPP1 suppressed the malignant biological behavior of hepatoma cells and tumor growth by upregulating miR-582-5p and downregulating KPNA2.

Objective:To evaluate the expression of B7-H3 and CD133 in colorectal cancer(CRC),colorectal polyps,and normal colorectal mucosa and investigate their roles in the development and prognosis of CRC.Methods:Immunohistochemistry was used to detect the ex-pression of B7-H3 and CD133 in 195 CRC,76 villous/tubulovillous adenoma,64 tubular adenoma,30 non-adenomatous polyp,and 10 nor-mal colorectal mucosa samples obtained from The Second Affiliated Hospital of Zhengzhou University between October 2012 and April 2017 and Pengan County People's Hospital between January 2017 and May 2019.Patient age,sex,and immunohistochemical staining results of B7-H3,CD133,and carcinoembryonic antigen were incorporated as risk factors to establish a CRC survival prediction model.Results:B7-H3 and CD133 expression showed an increasing trend from normal mucosa to non-adenomatous polyps,tubular adenomas,villous/tubulovil-lous adenomas,and CRC(P<0.05),and correlated with adenoma size.It was also associated with CRC metastasis and shorter survival(P<0.05).Furthermore,the expressions of B7-H3 and CD133 demonstrated a value in the CRC survival prediction model,in the training as well as validation set.Conclusions:The immune regulator B7-H3 and cancer stem cell marker CD133 are associated with poor prognosis in CRC,and their expressions may serve as predictive factors for CRC prognosis.