BACKGROUND:Studies have shown that intradiscal injection of syringin solution can improve the structure and function of the intervertebral disc,prevent and slow down the process of intervertebral disc degeneration in rats.However,the biological half-life of syringin is short and it is difficult for it to continue to play a role in the intervertebral disc.Its bioavailability needs to be further improved.OBJECTIVE:To observe the effect of syringin-chitosan hydrogel on intervertebral disc degeneration in rats and the mechanism of syringin in the treatment of intervertebral disc degeneration.METHODS:(1)Cell experiment:Passages 2-5 rat nucleus pulposus cells were divided into four groups for treatment.The normal control group did not undergo any treatment.The degeneration group was added with interleukin-1β(to establish the intervertebral disc degeneration cell model).The drug group was added with interleukin-1β and syringing.The inhibitor group was added with interleukin-1β,syringing,and phosphatidylinositol 3-kinase(PI3K)inhibitor LY294002.After 24 hours of treatment,apoptosis,extracellular matrix,oxidative stress,and apoptosis-related proteins and PI3K/protein kinase B(AKT)signaling pathway proteins were detected respectively.(2)Animal experiment:Syringin-chitosan hydrogels were prepared,and the micromorphology and slow-release properties of the hydrogels were tested.Thirty SD rats were randomly divided into model control group,model intervention group,hydrogel group,syringin solution group,and syringin hydrogel group,with 6 rats in each group.The intervertebral disc degeneration model was established by the acupuncture method.Immediately after model establishment,the rats in model intervention group,hydrogel group,syringin solution group,and syringin hydrogel group were injected with PBS,chitosan hydrogel,syringin solution,and syringin-chitosan hydrogel,respectively.The samples were taken 8 weeks after modeling for histological detection.RESULTS AND CONCLUSION:(1)Cell experiment:Compared with the normal control group,apoptosis rate,reactive oxygen species level,and expression of BAX,cleaved caspase-9,cleaved caspase-3,and matrix metalloproteinase 13 protein were increased in the nucleus pulpocytes in the degeneration group(P＜0.05),and the expression levels of p-PI3K,p-AKT,BCL-2,and type Ⅱ collagen were decreased(P＜0.05).Superoxide dismutase activity decreased(P＜0.05).Compared with the degeneration group,apoptotic rate,reactive oxygen species level,and expression of BAX,cleaved caspase-9,cleaved caspase-3,and matrix metalloproteinase 13 protein were decreased in the syringin solution and syringin solution groups(P＜0.05),and expression levels of p-PI3K,p-AKT,BCL-2,and type Ⅱ collagen were increased(P＜0.05).Superoxide dismutase activity increased(P＜0.05).LY294002 could partially inhibit the effect of syringin.(2)Animal experiment:Syringin-chitosan hydrogel had a loose porous structure and good slow-release performance.Hematoxylin-eosin and safranin O-fast green staining showed that compared with the model control group and model intervention group,chitosan hydrogel,syringin solution and syringing-chitosan hydrogel could improve the intervertebral disc degeneration in different degrees,and the therapeutic effect of syringing-chitosan hydrogel was better than that of hydrogel and drug solution alone.(3)These findings indicate that syringin can regulate apoptosis of nucleus pulposus cells and extracellular matrix degradation induced by oxidative stress by activating PI3K/AKT signaling pathway,thus delaying disc degeneration.Compared with syringin injection alone,syringin loading in chitosan hydrogel can further delay the progression of intervertebral disc degeneration in rats.

BACKGROUND:Studies have shown that intradiscal injection of syringin solution can improve the structure and function of the intervertebral disc,prevent and slow down the process of intervertebral disc degeneration in rats.However,the biological half-life of syringin is short and it is difficult for it to continue to play a role in the intervertebral disc.Its bioavailability needs to be further improved.OBJECTIVE:To observe the effect of syringin-chitosan hydrogel on intervertebral disc degeneration in rats and the mechanism of syringin in the treatment of intervertebral disc degeneration.METHODS:(1)Cell experiment:Passages 2-5 rat nucleus pulposus cells were divided into four groups for treatment.The normal control group did not undergo any treatment.The degeneration group was added with interleukin-1β(to establish the intervertebral disc degeneration cell model).The drug group was added with interleukin-1β and syringing.The inhibitor group was added with interleukin-1β,syringing,and phosphatidylinositol 3-kinase(PI3K)inhibitor LY294002.After 24 hours of treatment,apoptosis,extracellular matrix,oxidative stress,and apoptosis-related proteins and PI3K/protein kinase B(AKT)signaling pathway proteins were detected respectively.(2)Animal experiment:Syringin-chitosan hydrogels were prepared,and the micromorphology and slow-release properties of the hydrogels were tested.Thirty SD rats were randomly divided into model control group,model intervention group,hydrogel group,syringin solution group,and syringin hydrogel group,with 6 rats in each group.The intervertebral disc degeneration model was established by the acupuncture method.Immediately after model establishment,the rats in model intervention group,hydrogel group,syringin solution group,and syringin hydrogel group were injected with PBS,chitosan hydrogel,syringin solution,and syringin-chitosan hydrogel,respectively.The samples were taken 8 weeks after modeling for histological detection.RESULTS AND CONCLUSION:(1)Cell experiment:Compared with the normal control group,apoptosis rate,reactive oxygen species level,and expression of BAX,cleaved caspase-9,cleaved caspase-3,and matrix metalloproteinase 13 protein were increased in the nucleus pulpocytes in the degeneration group(P＜0.05),and the expression levels of p-PI3K,p-AKT,BCL-2,and type Ⅱ collagen were decreased(P＜0.05).Superoxide dismutase activity decreased(P＜0.05).Compared with the degeneration group,apoptotic rate,reactive oxygen species level,and expression of BAX,cleaved caspase-9,cleaved caspase-3,and matrix metalloproteinase 13 protein were decreased in the syringin solution and syringin solution groups(P＜0.05),and expression levels of p-PI3K,p-AKT,BCL-2,and type Ⅱ collagen were increased(P＜0.05).Superoxide dismutase activity increased(P＜0.05).LY294002 could partially inhibit the effect of syringin.(2)Animal experiment:Syringin-chitosan hydrogel had a loose porous structure and good slow-release performance.Hematoxylin-eosin and safranin O-fast green staining showed that compared with the model control group and model intervention group,chitosan hydrogel,syringin solution and syringing-chitosan hydrogel could improve the intervertebral disc degeneration in different degrees,and the therapeutic effect of syringing-chitosan hydrogel was better than that of hydrogel and drug solution alone.(3)These findings indicate that syringin can regulate apoptosis of nucleus pulposus cells and extracellular matrix degradation induced by oxidative stress by activating PI3K/AKT signaling pathway,thus delaying disc degeneration.Compared with syringin injection alone,syringin loading in chitosan hydrogel can further delay the progression of intervertebral disc degeneration in rats.

OBJECTIVE To prepare and characterize curcumin nanomicelles （hereinafter referred to as Cur/mPEG-PBLA micelles）， and to evaluate the in vitro hepatoprotective activity against alcohol liver disease （ALD）. METHODS Cur/mPEG-PBLA micelles were prepared with the dialysis method using methoxy-poly（ethylene glycol）-poly（β-benzyl-L-aspartate） （mPEG-PLGA） as the carrier. The appearance and microscopic morphology of Cur/mPEG-PBLA micelles were observed， and particle size， polydispersity index， Zeta potential， encapsulation efficiency and drug loading content were all detected. The in vitro release， pH stability， thermal stability， dilution stability， storage stability， plasma stability tests， and hemolysis experiments were all performed. The cell model of ALD was established with anhydrous ethanol intervention using human liver cancer cells and normal liver cells as objects， Cur reference solution as reference， to evaluate in vitro preventive and ameliorative effects of Cur/mPEG- PBLA micelles on ALD. RESULTS The prepared Cur/mPEG-PBLA micelles exhibited a pale-yellow milky light， with a spherical shape and uniform distribution. The average particle size was about 140 nm， and the polydispersity index was less than 0.3. Zeta potential was （－8.15±0.05） mV； the encapsulation efficiency was （73.26±3.16）%， and the drug loading content was （4.87± 0.42）%. The cumulative release of Cur reference substance was close to 80% at 10 h； the cumulative release of Cur/mPEG-PBLA micelles at 8 h was 28.94% and only 48.25% at 48 h. pH stability and thermal stability of Cur/mPEG-PBLA micelles were better than those of Cur reference solution； Cur/mPEG-PBLA micelles showed good dilution stability， storage stability and plasma stability， and would not cause hemolysis. Cur reference solution and Cur/mPEG-PBLA micelles had varying degrees of in vitro preventive and ameliorative effects on ALD in two types of cells； after 48 h of application， the above effects of Cur/mPEG-PBLA micelles were significantly better than those of Cur reference solution at the same mass concentration （P＜0.05）. CONCLUSIONS Cur/mPEG-PBLA micelles can improve pH stability and thermal stability of Cur， delayits degradation rate， and have better in vitro hepatoprotective activity against ALD.

The proband was a 33-year-old pregnant woman (G4P1) who suffered spontaneous abortion in the first 3 months of pregnancy without a history of blood transfusion or transplantation. The fourth pregnancy was clinically diagnosed with threatened abortion, and a cesarean section was performed on June 28, 2023, at the Obstetrics and Gynecology Department of Dongguan Hospital of Traditional Chinese Medicine. During cross-matching tests, unexpected antibodies were detected in the proband′s plasma, which could not be specifically identified, and no suitable donor red blood cells could be found. The blood samples were sent to the Blood Transfusion Laboratory of Dongguan Blood Center. The laboratory used serology to identify the erythrocyte phenotype of the proband and confirmed the proband as having a rare p blood group. The unexpected antibody was identified as anti-PP1P K, and gene sequencing of the proband revealed that the new allele A4GALT* (c.100G>A+c.418_428delins) was homozygous, which is speculated to cause changes in the polypeptide chains p.Veral34ile and p.GERln140TRPFS *73, and inactivation of α1, 4-galactosyltransferase. At the same time, another new allele A4GALT*c.100G>A was found in family members, and it was predicted that the single change of p.Val34Ile caused by this mutation would not affect protein function or enzyme activity.

【Objective】 To explore the challenging blood cross-matching and resolution for multiple myeloma (MM) patients in different disease stages. 【Methods】 For a patient who was first diagnosed as MM and scheduled for blood transfusion, his blood was cross matched with donors’ blood by microcolumn gel method and tube test. When the major side of cross-matching was agglutinated, the patient’s plasma was cross matched with donors’ red blood cell (RBC) by polybrene test, then plasma dilution cross matched with donors’ RBC by microcolumn gel method. For a patient who was diagnosed as recurrent refractory MM and scheduled for blood transfusion, his blood was cross matched with donors’ blood by microcolumn gel method. 【Results】 1) Case 1 was a first-visit outpatient. The major side of microcolumn cross-match test was agglutinated with the shape of fine line. The result of tube method also showed agglutination of major sides, and the rouleaux were detected by the microscopy. Then polybrene method and microcolumn gel method (after plasma diluted) were applied for cross-matching again with the above two donors’ blood and showed compatibility. 2) Case 2 was a recurrent refractory MM patient. The major and minor sides of microcolumn cross-match test were both agglutinated with the shape of granular. The patient was treated with anti-CD38 monoclonal antibody. The RBCs, after treated with dithiothreitol (DTT) was used to cross match with patient plasma by microcolumn test, and the result was compatible. 【Conclusion】 Polybrene method and microcolumn gel method after plasma diluted are suitable for blood cross-matching of newly diagnosed MM patients, also for those treated with CD38 monoclonal antibody, as the drug interference with cross-matching can be eliminated by DTT.

A 38-year-old female patient with uraemia received tacrolimus 1 mg orally twice daily after kidney transplantation. The serum trough concentration of tacrolimus was 9.8 μg/L on the 15th day. On the 17th day, the patient developed abdominal pain, nausea, and vomiting. The abdominal CT showed that the volume of pancreas increased, the edge was irregular, and obvious exudation appeared around the pancreas and in the abdominal cavity. Laboratory tests showed blood amylase 430 U/L and lipase 231 U/L.The acute pancreatitis related to tacrolimus was considered. Tacrolimus was stopped and replaced by cyclosporine A 100 mg orally twice daily. Meanwhile, symptomatic and supportive treatments such as fasting, gastrointestinal decompression, lipid-lowering, acid inhibition, anti-infection, and parenteral nutrition were given. Three days later, the above symptoms were relieved. Nineteen days later, laboratory tests showed blood amylase 173 U/L and lipase 51 U/L; abdominal CT showed that the outline of the pancreas was clearer than before, and the exudation around was significantly reduced.

A 38-year-old female patient with uraemia received tacrolimus 1 mg orally twice daily after kidney transplantation. The serum trough concentration of tacrolimus was 9.8 μg/L on the 15th day. On the 17th day, the patient developed abdominal pain, nausea, and vomiting. The abdominal CT showed that the volume of pancreas increased, the edge was irregular, and obvious exudation appeared around the pancreas and in the abdominal cavity. Laboratory tests showed blood amylase 430 U/L and lipase 231 U/L.The acute pancreatitis related to tacrolimus was considered. Tacrolimus was stopped and replaced by cyclosporine A 100 mg orally twice daily. Meanwhile, symptomatic and supportive treatments such as fasting, gastrointestinal decompression, lipid-lowering, acid inhibition, anti-infection, and parenteral nutrition were given. Three days later, the above symptoms were relieved. Nineteen days later, laboratory tests showed blood amylase 173 U/L and lipase 51 U/L; abdominal CT showed that the outline of the pancreas was clearer than before, and the exudation around was significantly reduced.

Urea transporters (UT) play a vital role in the mechanism of urine concentration and are recognized as novel targets for the development of salt-sparing diuretics. Thus, UT inhibitors are promising for development as novel diuretics. In the present study, a novel UT inhibitor with a diarylamide scaffold was discovered by high-throughput screening. Optimization of the inhibitor led to the identification of a promising preclinical candidate,

Objective: To investigate the effects and potential mechanisms of tolvaptan on chronic intermittent hypoxia (CIH)-induced atrial remodeling in rats. Methods: A total of 45 Sprague-Dawley rats were divided into 3 groups by the random number table: control group, CIH group (6 h/d for 30 days), CIH plus tolvaptan group (8 mg·kg^-1·d^-1 per gavage for 30 days). Echocardiography examination was performed after 30 days. Thereafter, 5 rats were randomly chosen for histology evaluation, 5 for molecular biological examinations and another 5 rats underwent isolated heart electrophysiology study in each group. Protein and mRNA expression levels of miRNA-21, Spry1, PTEN, ERK/p-ERK, MMP-9, PI3K, AKT/p-AKT were detected. Results: Compared to the rats in control group, rats in the CIH group showed higher atrial interstitial collagen deposition (P<0.001), increased atrial fibrillation inducibility (P=0.022). The results of immunohistochemistry staining showed that the mean optical density (MOD) of ERK, p-ERK and MMP-9 were significantly increased (all P<0.05), the MOD of Spry1 and PTEN were significantly decreased (both P<0.05), above changes could be significantly reversed by cotreatment with tolvaptan. No significant differences were detected in PI3K and AKT among the three groups (P>0.05). In addition, compared with rats in control group, mRNA levels of miRNA-21, MMP-9, PI3K, AKT, and protein levels of ERK, p-ERK, MMP-9 were significantly increased in CIH group(all P<0.05), whereas protein levels of Spry1, PI3K, p-AKT were significantly decreased (all P<0.05). Above changes could be significantly attenuated. Conclusions CIH induces significant atrial remodeling in this rat model, which can be attenuated by tolvaptan possibly through modulating miRNA-21/Spry1/ERK/MMP-9 and miRNA-21/PTEN/PI3K/AKT signaling pathways.

Objective To investigate the effects and potential mechanisms of tolvaptan on chronic intermittent hypoxia (CIH)?induced atrial remodeling in rats. Methods A total of 45 Sprague?Dawley rats were divided into 3 groups by the random number table: control group, CIH group (6 h/d for 30 days), CIH plus tolvaptan group (8 mg·kg-1·d-1 per gavage for 30 days). Echocardiography examination was performed after 30 days. Thereafter, 5 rats were randomly chosen for histology evaluation, 5 for molecular biological examinations and another 5 rats underwent isolated heart electrophysiology study in each group. Protein and mRNA expression levels of miRNA?21, Spry1, PTEN, ERK/p?ERK, MMP?9, PI3K, AKT/p?AKT were detected. Results Compared to the rats in control group, rats in the CIH group showed higher atrial interstitial collagen deposition (P<0.001), increased atrial fibrillation inducibility (P=0.022). The results of immunohistochemistry staining showed that the mean optical density (MOD) of ERK, p?ERK and MMP?9 were significantly increased (all P<0.05), the MOD of Spry1 and PTEN were significantly decreased (both P<0.05), above changes could be significantly reversed by cotreatment with tolvaptan. No significant differences were detected in PI3K and AKT among the three groups (P>0.05). In addition, compared with rats in control group, mRNA levels of miRNA?21, MMP?9, PI3K, AKT, and protein levels of ERK, p?ERK, MMP?9 were significantly increased in CIH group(all P<0.05), whereas protein levels of Spry1, PI3K, p?AKT were significantly decreased (all P<0.05). Above changes could be significantly attenuated. Conclusions CIH induces significant atrial remodeling in this rat model, which can be attenuated by tolvaptan possibly through modulating miRNA?21/Spry1/ERK/MMP?9 and miRNA?21/PTEN/PI3K/AKT signaling pathways.