ObjectiveTo investigate the effect of the herb pair Cuscutae Semen-Lycii Fructus on oxidative stress-induced blood-testis barrier dysfunction and spermatogenesis in the rat model of oligoasthenozoospermia （OAS） and decipher the mechanism based on the silent information regulator 1 （SIRT1）/nuclear factor erythroid 2-related factor 2 （Nrf2） signaling pathway. MethodsThirty-five male SD rats were randomized into a blank group （n=7） and a modeling group （n=28）. The OAS model was established by gavage of hydrocortisone aqueous solution combined with single factor electrical stimulation. The modeled rats were randomly assigned into the following groups： model， Cuscutae Semen-Lycii Fructus granules （3.2 g·kg^-1）， Cuscutae Semen-Lycii Fructus total flavonoids （0.34 g·kg^-1）， and L-carnitine （0.38 g·kg^-1）， and treated for 4 weeks. The sperm quality of rats was assessed by an automatic sperm analyzer. The levels of superoxide dismutase （SOD）， malondialdehyde （MAD）， and glutathione peroxidase （GSH-Px） in the testicular tissue were determined by enzyme-linked immunosorbent assay. Hematoxylin-eosin staining was employed to reveal the pathological changes in the testicular tissue and score the spermatogenic function. Transmission electron microscopy was employed to observe the ultrastructural changes of Sertoli cells. Western blot and Real-time PCR were employed to determine the protein and mRNA levels， respectively， of SIRT1， Nrf2， Occludin， zonula occludens-1 （ZO-1）， connexin 43 （CX43）， and β-catenin. ResultsCompared with the blank group， the model group showed decreased total sperm count and motility （P<0.05， P<0.01）， obvious damage in the testicular tissue and blood-testis barrier structure， reduced score of spermatogenic function （P<0.01）， declined levels of GSH-Px and SOD in the testicular tissue （P<0.05）， elevated level of MDA， and down-regulated protein levels of SIRT1， Nrf2， ZO-1， CX43， β-catenin， and occludin （P<0.05， P<0.01） and mRNA levels of SIRT1， Nrf2， ZO-1， CX43， and β-catenin in the testicular tissue （P<0.05， P<0.01）. After treatment， the testicular tissue， blood-testis barrier structure， and score of spermatogenic function （P<0.01） were improved in the Cuscutae Semen-Lycii Fructus granules group， Cuscutae Semen-Lycii Fructus total flavonoids group， and L-carnitine group. Compared with the model group， the treatment groups presented lowered levels of GSH-Px and SOD （P<0.05， P<0.01）， and the Cuscutae Semen-Lycii Fructus granule group showed a decline in MDA level. The protein and mRNA levels of SIRT1， Nrf2， ZO-1， CX43， β-catenin， and occludin were up-regulated in the Cuscutae Semen-Lycii Fructus granules group and total flavonoids group （P<0.05， P<0.01）. ConclusionThe herb pair Cuscutae Semen-Lycii Fructus can regulate the SIRT1/Nrf2 pathway to inhibit oxidative stress and alleviate the blood-testis barrier damage， thereby improving the spermatogenic function in the rat model of OAS. Total flavonoids may be the material basis for the therapeutic effect of Cuscutae Semen-Lycii Fructus.

ObjectiveTo investigate the effect of the herb pair Cuscutae Semen-Lycii Fructus on oxidative stress-induced blood-testis barrier dysfunction and spermatogenesis in the rat model of oligoasthenozoospermia （OAS） and decipher the mechanism based on the silent information regulator 1 （SIRT1）/nuclear factor erythroid 2-related factor 2 （Nrf2） signaling pathway. MethodsThirty-five male SD rats were randomized into a blank group （n=7） and a modeling group （n=28）. The OAS model was established by gavage of hydrocortisone aqueous solution combined with single factor electrical stimulation. The modeled rats were randomly assigned into the following groups： model， Cuscutae Semen-Lycii Fructus granules （3.2 g·kg^-1）， Cuscutae Semen-Lycii Fructus total flavonoids （0.34 g·kg^-1）， and L-carnitine （0.38 g·kg^-1）， and treated for 4 weeks. The sperm quality of rats was assessed by an automatic sperm analyzer. The levels of superoxide dismutase （SOD）， malondialdehyde （MAD）， and glutathione peroxidase （GSH-Px） in the testicular tissue were determined by enzyme-linked immunosorbent assay. Hematoxylin-eosin staining was employed to reveal the pathological changes in the testicular tissue and score the spermatogenic function. Transmission electron microscopy was employed to observe the ultrastructural changes of Sertoli cells. Western blot and Real-time PCR were employed to determine the protein and mRNA levels， respectively， of SIRT1， Nrf2， Occludin， zonula occludens-1 （ZO-1）， connexin 43 （CX43）， and β-catenin. ResultsCompared with the blank group， the model group showed decreased total sperm count and motility （P<0.05， P<0.01）， obvious damage in the testicular tissue and blood-testis barrier structure， reduced score of spermatogenic function （P<0.01）， declined levels of GSH-Px and SOD in the testicular tissue （P<0.05）， elevated level of MDA， and down-regulated protein levels of SIRT1， Nrf2， ZO-1， CX43， β-catenin， and occludin （P<0.05， P<0.01） and mRNA levels of SIRT1， Nrf2， ZO-1， CX43， and β-catenin in the testicular tissue （P<0.05， P<0.01）. After treatment， the testicular tissue， blood-testis barrier structure， and score of spermatogenic function （P<0.01） were improved in the Cuscutae Semen-Lycii Fructus granules group， Cuscutae Semen-Lycii Fructus total flavonoids group， and L-carnitine group. Compared with the model group， the treatment groups presented lowered levels of GSH-Px and SOD （P<0.05， P<0.01）， and the Cuscutae Semen-Lycii Fructus granule group showed a decline in MDA level. The protein and mRNA levels of SIRT1， Nrf2， ZO-1， CX43， β-catenin， and occludin were up-regulated in the Cuscutae Semen-Lycii Fructus granules group and total flavonoids group （P<0.05， P<0.01）. ConclusionThe herb pair Cuscutae Semen-Lycii Fructus can regulate the SIRT1/Nrf2 pathway to inhibit oxidative stress and alleviate the blood-testis barrier damage， thereby improving the spermatogenic function in the rat model of OAS. Total flavonoids may be the material basis for the therapeutic effect of Cuscutae Semen-Lycii Fructus.

Colorectal cancer （CRC） is a prevalent malignant tumor of the digestive tract， with a high incidence and high mortality. The majority of patients are diagnosed at the middle or advanced stage， which severely influences and threatens their physical health. Current treatment modalities such as surgery， radiotherapy， and chemotherapy often encounter challenges including metastasis， recurrence， and drug resistance. The phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway serves as a classical regulator that regulates physiological processes such as cell cycle， autophagy， apoptosis， and proliferation. Overexpression of this pathway is observed in various tumors. In the context of CRC， the activation of this pathway can facilitate the proliferation， invasion， and migration， inhibit the autophagy and apoptosis， promote the epithelial-mesenchymal transition of CRC cells， enhance angiogenesis within the tumor， and contribute to chemotherapy resistance and radiation resistance in CRC. Traditional Chinese medicine （TCM） treatment can exert an anti-CRC effect by inhibiting this pathway， thereby improving clinical efficacy and safety. This article retrieves relevant research literature published domestically and internationally regarding the regulation of the PI3K/Akt/mTOR signaling pathway by TCM in the treatment of CRC and conducts detailed classification and summary. The active components of TCM include glycosides， flavonoids， alkaloids， terpenoids， polyphenols， and naphthoquinones. The volatile oils and extracts of TCM include Angelicae Sinensis Radix volatile oil， Astragali Radix polysaccharides， Caryophylli Flos extract， Forsythiae Fructus extract， Curcumae Longae Rhizoma extract， and Celastrus orbiculatus extract. The compound formulas of TCM include Banxia Xiexin decoction， Jianpi Qingre Huoxue formula， and Chanling Plaster. Through summary and analysis， it is discovered that the abovementioned TCM can produce effects such as blocking the cell cycle， inducing autophagy and apoptosis， inhibiting angiogenesis， suppressing proliferation and migration， and reversing chemotherapy resistance and radiotherapy resistance by inhibiting the PI3K/Akt/mTOR pathway in CRC cells. TCM holds promise in the research and application of targeting the PI3K/Akt/mTOR signaling pathway for CRC treatment. The summary and conclusion of this article aim to provide references for subsequent research and the development of new drugs.

Colorectal cancer （CRC） is a prevalent malignant tumor of the digestive tract， with a high incidence and high mortality. The majority of patients are diagnosed at the middle or advanced stage， which severely influences and threatens their physical health. Current treatment modalities such as surgery， radiotherapy， and chemotherapy often encounter challenges including metastasis， recurrence， and drug resistance. The phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） signaling pathway serves as a classical regulator that regulates physiological processes such as cell cycle， autophagy， apoptosis， and proliferation. Overexpression of this pathway is observed in various tumors. In the context of CRC， the activation of this pathway can facilitate the proliferation， invasion， and migration， inhibit the autophagy and apoptosis， promote the epithelial-mesenchymal transition of CRC cells， enhance angiogenesis within the tumor， and contribute to chemotherapy resistance and radiation resistance in CRC. Traditional Chinese medicine （TCM） treatment can exert an anti-CRC effect by inhibiting this pathway， thereby improving clinical efficacy and safety. This article retrieves relevant research literature published domestically and internationally regarding the regulation of the PI3K/Akt/mTOR signaling pathway by TCM in the treatment of CRC and conducts detailed classification and summary. The active components of TCM include glycosides， flavonoids， alkaloids， terpenoids， polyphenols， and naphthoquinones. The volatile oils and extracts of TCM include Angelicae Sinensis Radix volatile oil， Astragali Radix polysaccharides， Caryophylli Flos extract， Forsythiae Fructus extract， Curcumae Longae Rhizoma extract， and Celastrus orbiculatus extract. The compound formulas of TCM include Banxia Xiexin decoction， Jianpi Qingre Huoxue formula， and Chanling Plaster. Through summary and analysis， it is discovered that the abovementioned TCM can produce effects such as blocking the cell cycle， inducing autophagy and apoptosis， inhibiting angiogenesis， suppressing proliferation and migration， and reversing chemotherapy resistance and radiotherapy resistance by inhibiting the PI3K/Akt/mTOR pathway in CRC cells. TCM holds promise in the research and application of targeting the PI3K/Akt/mTOR signaling pathway for CRC treatment. The summary and conclusion of this article aim to provide references for subsequent research and the development of new drugs.

Most chemical medicines have polymorphs. The difference of medicine polymorphs in physicochemical properties directly affects the stability, efficacy, and safety of solid medicine products. Polymorphs is incomparably important to pharmaceutical chemistry, manufacturing, and control. Meantime polymorphs is a key factor for the quality of high-end drug and formulations. Polymorph prediction technology can effectively guide screening of trial experiments, and reduce the risk of missing stable crystal form in the traditional experiment. Polymorph prediction technology was firstly based on theoretical calculations such as quantum mechanics and computational chemistry, and then was developed by the key technology of machine learning using the artificial intelligence. Nowadays, the popular trend is to combine the advantages of theoretical calculation and machine learning to jointly predict crystal structure. Recently, predicting medicine polymorphs has still been a challenging problem. It is expected to learn from and integrate existing technologies to predict medicine polymorphs more accurately and efficiently.

Objective To compare the blood glucose control effect and safety between insulin degludec and insulin aspart and insulin aspart 30 in non-obese patients with type 2 diabetes mellitus(T2DM).Methods Non-obese patients with T2DM were divided into treatment group and control group according to different treatment methods.The control group was treated with insulin aspart 30,and the treatment group was treated with insulin degludec and insulin aspart.Pancreatic islet related indicators[fasting C-peptide(FCP)and 2-hour postprandial C-peptide(2 h CP)],blood glucose control effect[fasting blood glucose(FBG),2-hour postprandial blood glucose(2 h PG)and glycated hemoglobin(HbA1c)],serum 25-hydroxyvitamin D[25(OH)D]and the risk of hypoglycemia were compared between the two groups.Results There were 41 cases in treatment group and 39 cases in control group.After treatment,FCP levels in treatment group and control group were(0.84±0.09)and(1.07±0.14)nmol·L-1;2 h CP levels were(1.03±0.15)and(1.69±0.17)nmol·L-1;FBG levels were(5.46±0.57)and(6.18±0.67)mmol·L-1;2 h PG levels were(8.17±0.85)and(9.03±0.94)mmol·L-1;HbA1 c were(5.35±0.57)％and(6.47±0.68)％;25(OH)D levels were(26.33±2.75)and(20.54±2.17)nmol·L-1,all with significant difference(all P＜0.05).The incidence rates of non-severe hypoglycemia in treatment group and control group were 14.63％and 35.90％,with statistically significant difference(P＜0.05).The incidence rates of severe hypoglycemia in treatment group and control group were 9.76％and 12.82％;the incidence rates of nocturnal hypoglycemia were 19.51％and 17.95％,without statistically significant difference(all P＞0.05).Conclusion The overall therapeutic effect of insulin degludec and insulin aspart on non-obese patients with T2DM is better than that of insulin aspart 30.The former can effectively regulate blood glucose and pancreatic islet cell function,lower the risk of non-severe hypoglycemia.

Objective To explore the effect of Maixuekang capsules on renal fibrosis and inhibition of Janus tyrosine protein kinase 2(JAK2)/signal transducer and activator of transcription family 3(STAT3)signal pathway in rats with immunoglobulin A nephropathy(IgAN).Methods The rat model of IgAN was established by modified 600 mg·kg-1 BSA+LPS 2 mL+CCL4 0.1 mL scheme,and the 24-hour urinary protein was detected.Fifty-five SD rats were randomly divided into 6 groups:Blank control group(distilled water+0.9％NaCl),model group(IgAN model),positive control group(10 mg·kg-1 telmisartan tablets after modeling),experimental-L,-M,-H groups(16,44,84 ATU·kg-1 Maixuekang capsules after modeling).The levels of serum creatinine(Scr),blood urea nitrogen(BUN),glutamic pyruvic transaminase(GPT)and glutamic oxalacetic transaminase(GOT)were determined by automatic biochemical analyzer.The morphology of kidney was observed by hematoxylin-eosin(HE)staining,and the deposition of IgA in rat kidney was detected by immunofluorescence.The expressions of α-SMA,TGF-β1,JAK2,STAT3 and P-STAT3 in renal tissue were detected by Western blot.Results The blood Scr of blank control group,model group,positive control and experimental-H groups were(41.42±2.30),(59.82±5.36),(50.28±3.18)and(50.47±7.19)μ mol·L-1,respectively;the 24-hour urinary protein were(2.35±1.21),(11.62±3.45),(5.01±4.36)and(8.14±4.19)mg·d-1,respectively;the relative expression of α-SMA protein were 0.48±0.10,1.00±0.00,0.71±0.13 and 0.57±0.22,respectively;the relative expression of TGF-β1 protein were 0.54±0.93,1.00±0.00,0.73±0.33 and 0.51±0.31,respectively;the relative expression of JAK2 protein were 0.36±0.22,1.00±0.00,0.82±0.36 and 0.57±0.28,respectively;p-STAT3/STAT3 were 0.40±0.19,1.00±0.00,0.46±0.18 and 0.31±0.18,respectively.Compared with the blank control group,the differences of above indexes in the model group were statistically significant(all P＜0.05).Compared with the model group,the differences of above indexes in the positive control and experimental-H groups were statistically significant(all P＜0.05).Conclusion Maixuekang capsules may reduce the level of renal fibrosis and protect renal function in rats with IgAN by inhibiting the activation of JAK2/STAT3 signal pathway.

Pyroptosis, an inflammatory caspase-dependent programmed cell death, plays a vital role in maintaining tissue homeostasis and activating inflammatory responses. Orthodontic tooth movement (OTM) is an aseptic force-induced inflammatory bone remodeling process mediated by the activation of periodontal ligament (PDL) progenitor cells. However, whether and how force induces PDL progenitor cell pyroptosis, thereby influencing OTM and alveolar bone remodeling remains unknown. In this study, we found that mechanical force induced the expression of pyroptosis-related markers in rat OTM and alveolar bone remodeling process. Blocking or enhancing pyroptosis level could suppress or promote OTM and alveolar bone remodeling respectively. Using Caspase-1^-/- mice, we further demonstrated that the functional role of the force-induced pyroptosis in PDL progenitor cells depended on Caspase-1. Moreover, mechanical force could also induce pyroptosis in human ex-vivo force-treated PDL progenitor cells and in compressive force-loaded PDL progenitor cells in vitro, which influenced osteoclastogenesis. Mechanistically, transient receptor potential subfamily V member 4 signaling was involved in force-induced Caspase-1-dependent pyroptosis in PDL progenitor cells. Overall, this study suggested a novel mechanism contributing to the modulation of osteoclastogenesis and alveolar bone remodeling under mechanical stimuli, indicating a promising approach to accelerate OTM by targeting Caspase-1.
Animals ; Humans ; Mice ; Rats ; Bone Remodeling/physiology* ; Caspase 1 ; Periodontal Ligament ; Pyroptosis ; Tooth Movement Techniques

【Objective】 To establish the internal quality control standard of leukocyte-depleted suspended red blood cell volume in our center, so as to guide the preparation of components, strengthen the internal quality control and improve the quality of blood preparations. 【Methods】 A total of 1 523 bags of whole blood collected using two manufacturers′ leukocyte-depleted blood bags from March to August 2023 at our center were extracted. The blood before and after filtration were weighed, and the volume of whole blood collected, the volume of filtration loss and the product volume based on the formula and measured specific gravity were calculated. According to the data distribution characteristics, the reference range of leukocyte-depleted suspended red blood cell volume was determined, and the differences of whole blood collection volume, filtration loss capacity and product capacity between the two manufacturers were analyzed. The quality control data of leukocyte-depleted red blood cells over the past year with the difference between another 100 bags of these cells and the reference interval were compared, and the effectiveness of reference interval was validated. 【Results】 The median whole blood collection volume in the sample size was 402.0 mL, with a median filtration loss of 42.4 mL, and an average volume of leukocyte-depleted suspended red blood cells at 322.5 mL. The whole blood collection volume (A: median 404.4 mL; B: median 397.7 mL, P<0.01) and the volume of leukocyte-depleted suspended red blood cell products (A: mean 331.4 mL; B: mean 312.0 mL, P<0.01) using manufacturer A′s leukocyte-depleted blood bag were both higher, with a lower filtration loss capacity (A: median 39.5 mL; B: median 46.6 mL, P<0.01). The standard deviation of the volume of leukocyte-depleted suspended red blood cell was 19.6, and the reference interval was 284.1-360.9 mL. The validation samples and quality control sampling data showed no difference from the interval samples (P>0.05). 【Conclusion】 According to the actual situation of our center, the volume standard of leukocyte-depleted suspended red blood cells in our center is determined to be 284.1-360.9 mL.

Pyroptosis is a form of lytic programmed cell death executed by a family of pore-forming proteins named gasdermin (GSDM). Pyroptosis plays crucial roles in host defense against pathogen infection and eliminating abnormal and harmful cells, while excessive pyroptosis causes inflammatory diseases including cytokine storm and septic shock. Mammalian GSDMs, except for pejvakin (PJVK), adopt an autoinhibited two-domain architecture, in which the N-terminal cytotoxic domain (GSDM-N) is restrained in an inactive state by the intramolecular interaction with the C-terminal inhibitory domain (GSDM-C). These two-domain proteins are activated by upstream protease cleavage within the interdomain linkers. The unleashed GSDM-N binds to acidic phospholipids in the cytoplasmic leaf of plasma membranes and undergoes dramatic conformational changes and oligomerization, then assembling into transmembrane pores for pyroptosis induction. GSDM pores lead to membrane rupture, cell swelling, and cytosol release, thereby mobilizing proinflammatory responses. GSDMs are evolutionarily conserved and have been discovered across all kingdoms of life, including bacteria, fungi, invertebrates such as cnidarians and mollusks, and all vertebrates. Proteolytic cleavage to liberate the pore-forming activity of GSDM-N appears to be a universal mechanism for most GSDMs activation, despite low sequence homology among the GSDMs from diverse species. However, recent studies discover that there exist noncanonical GSDMs lack of functional C-terminal inhibitory domains in some lower eukaryotic species. These noncanonical GSDMs are activated by unprecedent mechanisms independent of proteolytic cleavage. TrichoGSDM, present in the basal metazoan Trichoplax adhaerens, is a pore-forming domain-only protein and exists as a disulfides-linked autoinhibited dimer. Reduction of the disulfides by the conserved cytoplasmic antioxidant system, including glutathione (GSH) and thioredoxin (Trx), generates pore-forming active monomers capable of inducing lytic cell death. In filamentous fungus Neurospora crassa, polymorphic regulator of cell death-1 (rcd-1) encodes two GSDM-like proteins RCD-1-1 and RCD-1-2 in incompatible haplostrains, which trigger pyroptosis-like cell death in nonself discrimination (allorecognition) upon encountering during somatic cell fusion. RCD-1-1 and RCD-1-2 are both monomers and structurally similar to mammalian GSDM-N domains, lacking autoinhibitory fragments. They alone could bind acidic phospholipids, and associate with cell membrane in a resting state. Coexistence of RCD-1-1 and RCD-1-2 leads to formation of RCD-1-1/RCD-1-2 heterodimers through molecular mating, which further oligomerize into membrane-inserted pores, causing rapid lytic cell death. These findings reveal mechanistic diversities in GSDM activation and indicate versatile functions of GSDMs. Due to the highly proinflammatory nature of pyroptosis, the pore-forming activities of GSDMs have been illustrated to be precisely regulated at multiple levels. GSDMD transcription and expression is characterized to be induced by interferon regulatory factors 2 (IRF2). mRNA alternative splicing of GSDMB generates various isoforms, some of which exhibit potent pore-forming activity whereas the others bear none. Additionally, different types of post-translational modifications have been identified on GSDMs, playing distinct regulatory roles. For examples, itaconation of GSDMD, succinylation of GSDMD and GSDME, and phosphorylation of GSDMA, GSDMD and GSDME, negatively regulate GSDM pore formation, thereby inhibiting pyroptosis. Conversely, palmitoylation of GSDMD and GSDME, and ubiquitination of GSDMD promote the pore-forming activities and pyroptosis. Moreover, some proteases can cleave within the GSDM-N domains to block their pore-forming activities. On the other hand, bacterial pathogens evolve specific effectors to hijack host pyroptotic defense pathway through targeting upstream caspases, GSDMs or plasma membrane phospholipids. Given the crucial roles of GSDMD in immune defense and pathological inflammation, a few small-molecule inhibitors have been found to directly inhibit GSDMD activity. Since the identification of GSDMs as the executioners of pyroptosis, the GSDM family has attracted broad attention in immunology researches. Significant progress has been made to greatly advance our knowledge about how GSDMs action, and what are the immunological functions of pyroptosis. Investigations of GSDM-targeting therapies are emerging as a promising translational direction. In this paper, we review recent progress in the field of pyroptosis researches, with focus on various molecular mechanisms underlying GSDMs activation and regulation. The biological implication and future direction of pyroptosis research are also discussed.