Background and Aims:Hepatocellular carcinoma(HCC)is one of the most common malignancies,with high incidence and mortality.The JAK/STAT signaling pathway plays a crucial role in regulating cell proliferation,apoptosis,and immune responses,and its persistent activation is closely associated with the development of HCC.This study aimed to investigate the effects of the JAK2 inhibitor AG490 and the STAT3-related inhibitor rapamycin(RPM)on the biological behaviors of HCC cells and their underlying molecular mechanisms.Methods:Human hepatoma HepG2 cells were divided into four groups:blank control,AG490,RPM,and AG490+RPM.Cell proliferation,migration,and apoptosis were assessed by MTT assay,scratch test,and flow cytometry,respectively.ELISA and qRT-PCR were used to detect the protein and mRNA expression of Bax,Bcl-2,caspase-3,survivin,and c-Myc.A Western blot analysis was performed to examine the expression and phosphorylation levels of JAK2 and STAT3.Results:Compared with the blank control group,the AG490,RPM,and AG490+RPM groups showed significantly decreased cell proliferation and migration abilities,as well as increased apoptosis,with the most pronounced effects observed in the AG490+RPM group(all P＜0.05).Inhibitor-treated groups showed elevated expression of Bax and caspase-3,decreased expression of Bcl-2,survivin,and c-Myc,with the most significant changes in the AG490+RPM group(all P＜0.05).In addition,the ratios of p-JAK2/JAK2 and p-STAT3/STAT3 were significantly reduced in all treated groups,with the lowest levels in the AG490+RPM group(all P＜0.05).Conclusion:Inhibitors targeting the JAK/STAT pathway significantly suppress proliferation and migration,and induce apoptosis in HCC cells,possibly by downregulating p-JAK2 and p-STAT3,as well as modulating genes related to apoptosis and proliferation.The combined use of AG490 and RPM exhibits superior antitumor effects,suggesting that multi-target blockade of the JAK/STAT pathway may represent a promising therapeutic strategy for HCC.

Background and Aims:Hepatocellular carcinoma(HCC)is one of the most common malignancies,with high incidence and mortality.The JAK/STAT signaling pathway plays a crucial role in regulating cell proliferation,apoptosis,and immune responses,and its persistent activation is closely associated with the development of HCC.This study aimed to investigate the effects of the JAK2 inhibitor AG490 and the STAT3-related inhibitor rapamycin(RPM)on the biological behaviors of HCC cells and their underlying molecular mechanisms.Methods:Human hepatoma HepG2 cells were divided into four groups:blank control,AG490,RPM,and AG490+RPM.Cell proliferation,migration,and apoptosis were assessed by MTT assay,scratch test,and flow cytometry,respectively.ELISA and qRT-PCR were used to detect the protein and mRNA expression of Bax,Bcl-2,caspase-3,survivin,and c-Myc.A Western blot analysis was performed to examine the expression and phosphorylation levels of JAK2 and STAT3.Results:Compared with the blank control group,the AG490,RPM,and AG490+RPM groups showed significantly decreased cell proliferation and migration abilities,as well as increased apoptosis,with the most pronounced effects observed in the AG490+RPM group(all P＜0.05).Inhibitor-treated groups showed elevated expression of Bax and caspase-3,decreased expression of Bcl-2,survivin,and c-Myc,with the most significant changes in the AG490+RPM group(all P＜0.05).In addition,the ratios of p-JAK2/JAK2 and p-STAT3/STAT3 were significantly reduced in all treated groups,with the lowest levels in the AG490+RPM group(all P＜0.05).Conclusion:Inhibitors targeting the JAK/STAT pathway significantly suppress proliferation and migration,and induce apoptosis in HCC cells,possibly by downregulating p-JAK2 and p-STAT3,as well as modulating genes related to apoptosis and proliferation.The combined use of AG490 and RPM exhibits superior antitumor effects,suggesting that multi-target blockade of the JAK/STAT pathway may represent a promising therapeutic strategy for HCC.

BACKGROUND:There is an internal relationship between hyperhomocysteinemia and vascular calcification.However,the pathogenesis of hyperhomocysteinemia promoting vascular calcification is still unclear. OBJECTIVE:To investigate the role of bone morphogenetic protein-2 in hyperhomocysteinemia-induced vascular calcification. METHODS:Human carotid wax samples were divided into a calcified group(n=29)and a non-calcified group(n=13)according to the presence or absence of calcified plaque.Sixteen ApoE-/-mice were randomly divided into a control group and a hyperhomocysteinemia group,with 8 mice in each group.Bone morphogenetic protein-2 vector was used to transfect rat thoracic artery smooth muscle A7r5 cells,and gradient concentration of homocysteine(50,100,200,and 400 μmol/L)was utilized to treat A7r5 cells.Calcification was detected by alizarin red staining and hematoxylin-eosin staining.The interaction of bone morphogenetic protein 2 with Runt-related transcription factor 2 was detected by immunofluorescence,and the expressions of bone morphogenetic protein 2,Runt-related transcription factor 2,and α-smooth muscle actin were detected by immunohistochemistry and western blot assay. RESULTS AND CONCLUSION:(1)Human carotid artery tissue staining revealed that compared with the non-calcification group,inflammatory cells increased and calcification positive rate increased in the calcification group(P<0.05).Compared with the non-calcification group,the expressions of bone morphogenetic protein-2 and Runt-related transcription factor 2 were up-regulated,and the expression of α-smooth muscle actin was decreased in the calcification group(all P<0.05).(2)The staining of mouse arterial specimens exhibited that,the positive rate of calcified area in the hyperhomocysteinemia group was significantly higher than that in the control group(P<0.05);serum homocysteine level in the hyperhomocysteinemia group was significantly higher than that in the control group(P<0.05).Compared with the control group,the expressions of bone morphogenetic protein-2 and Runt-related transcription factor 2 were up-regulated,and the expression of α-smooth muscle actin was decreased in the hyperhomocysteinemia group(all P<0.05).(3)A7r5 cell culture analysis demonstrated that with the increase of homocysteine concentration gradient,the degree of calcification,the content of bone morphogenetic protein-2 and Runt-related transcription factor 2 protein in A7r5 cells increased(P<0.05),and the content of α-smooth muscle actin protein decreased(P<0.05).(4)The A7r5 cell culture analysis of overexpressed bone morphogenetic protein 2 showed that the calcification degree of the overexpressed bone morphogenetic protein 2 group was increased compared with the corresponding control group,the β-sodium glycerophosphate group,and the homocysteine group.RUNt-related transcription factor 2 expression up-regulated(P<0.05)and α-smooth muscle actin expression down-regulated(P<0.05).(5)The expression of bone morphogenetic protein 2 increased in A7r5 cells cultured with homocysteine in calcified medium,and the expression of Runt-related transcription factor 2 increased with the increase of bone morphogenetic protein 2 expression.(6)The results confirm that bone morphogenetic protein-2 is a key target gene in the regulation of smooth muscle cell phenotypic transformation resulting in vascular calcification by hyperhomocysteinemia.Targeted regulation of bone morphogenetic protein-2 reduces hyperhomocysteinemia-induced vascular calcification.

Objective:To investigate the protective effect of calycosin on microgravity-induced muscle atrophy by using real-time shear wave elastography (RT-SWE).Methods:The potential key active compound calycosin of anti-muscular atrophy in Astragali Radix was screened by systematic pharmacology. Eighteen healthy male Sprague-Dawley rats were divided into the control group [0.5% carboxymethyl cellulose-Na (CMC-Na) gavage], the hind limb unloading (HLU)+CMC-Na group (HLU+0.5% CMC-Na gavage), and the HLU+calycosin group (HLU+calycosin gavage) according to the method of random number table, with 6 rats in each group. After 28 d of continuous administration, the organ index, the toxicity of liver and kidney, the wet weight of soleus muscle and the ratio of muscle weight to body weight was measured, respectively. The non-invasive RT-SWE was used to evaluate the thickness and elastic modulus of rectus femoris in each group and the one-way analysis of variance was used to compare the differences among groups.Results:There was no significant difference in organ index, liver and kidney toxicity among different groups of rats (all P>0.05). There were significant differences in the weight of soleus muscle and the ratio of muscle weight to body weight among different groups of rats ( F=60.66, 56.44, both P<0.001). Compared with the HLU+CMC-Na group, the weight of soleus muscle and the ratio of muscle weight to body weight in the HLU+calycosin group increased, and the differences were significant (both P<0.01). The thickness and elastic modulus of rectus femoris of rats in different groups were significantly different ( F=35.47, 14.68, both P<0.001). Compared with the HLU+CMC-Na group, the muscle thickness and elastic modulus of rats in HLU+calycosin group were increased, and the differences were significant (both P<0.01). Conclusions:The treatment of calycosin has no side effects on rats. It can improve the thickness and elastic modulus of rectus femoris, and effectively prevent microgravity-induced muscle atrophy, which may provide a new candidate drug for astronaut muscular atrophy.

【Objective】 To analyze the related factors of emotional and behavioral abnormalities in children with overactive bladder (OAB). 【Methods】 OAB children (aged 6 to 16 years) in a survey of 5 032 children from a county in Henan Province during Sep.2022 and Dec.2022 were identified and surveyed with Overactive Bladder Symptom Score (OABSS), Strength and Difficulties Questionnaire (SDQ) and Pediatric Sleep Questionnaire (PSQ). According to the SDQ score, they were divided into abnormal group (SDQ≥20) and normal group. 【Results】 There were 35.7%(137/385) cases in the abnormal group and 64.3% (248/385) in the normal group. Gender, education level of caregivers, body mass index (BMI), age, constipation, enuresis and severity of OAB were significantly associated with emotional and behavioral abnormalities (P<0.05). Children in the abnormal group showed significant differences in emotional symptoms, conduct problems, hyperactivity symptoms, peer interaction and sleep (P<0.001). Multivariate regression analysis revealed significant differences in gender, educational level of caregi-vers, BMI, age, constipation, enuresis, severity of OAB and PSQI between the two groups (P<0.05). 【Conclusion】 The prevalence of emotional and behavioral abnormalities is high in children with OAB, which is related to female gender, high BMI, puberty, constipation, enuresis and severity of OAB.

Objective:To investigate the protective effect of calycosin on microgravity-induced muscle atrophy by using real-time shear wave elastography (RT-SWE).Methods:The potential key active compound calycosin of anti-muscular atrophy in Astragali Radix was screened by systematic pharmacology. Eighteen healthy male Sprague-Dawley rats were divided into the control group [0.5% carboxymethyl cellulose-Na (CMC-Na) gavage], the hind limb unloading (HLU)+CMC-Na group (HLU+0.5% CMC-Na gavage), and the HLU+calycosin group (HLU+calycosin gavage) according to the method of random number table, with 6 rats in each group. After 28 d of continuous administration, the organ index, the toxicity of liver and kidney, the wet weight of soleus muscle and the ratio of muscle weight to body weight was measured, respectively. The non-invasive RT-SWE was used to evaluate the thickness and elastic modulus of rectus femoris in each group and the one-way analysis of variance was used to compare the differences among groups.Results:There was no significant difference in organ index, liver and kidney toxicity among different groups of rats (all P>0.05). There were significant differences in the weight of soleus muscle and the ratio of muscle weight to body weight among different groups of rats ( F=60.66, 56.44, both P<0.001). Compared with the HLU+CMC-Na group, the weight of soleus muscle and the ratio of muscle weight to body weight in the HLU+calycosin group increased, and the differences were significant (both P<0.01). The thickness and elastic modulus of rectus femoris of rats in different groups were significantly different ( F=35.47, 14.68, both P<0.001). Compared with the HLU+CMC-Na group, the muscle thickness and elastic modulus of rats in HLU+calycosin group were increased, and the differences were significant (both P<0.01). Conclusions:The treatment of calycosin has no side effects on rats. It can improve the thickness and elastic modulus of rectus femoris, and effectively prevent microgravity-induced muscle atrophy, which may provide a new candidate drug for astronaut muscular atrophy.

Objective: To investigate the clinicopathologic characteristics, immunophenotypes, and differential diagnostic features of extra-pleural solitary fibrous tumor (SFT) with uncommon histology. Methods: Seven cases of extra-pleural SFT with uncommon histology were collected during January 2015 and December 2016 in Zhejiang Provincal People′s Hospital; the clinical and radiologic features, histomorphology, immunophenotype and prognosis were analyzed. EnVision method was used for immunohistochemical staining of STAT6, CD34 and other differential diagnosis associated markers. Results: There were five male and two female patients, age from 23 to 54 years (mean=39 years). Three tumors were located in the soft tissue of head and neck, two in trunk subcutaneous soft tissue, one in sella region, and one in the kidney. Grossly the tumors ranged from 0.4 to 8.0 cm (mean=3.1 cm). Microscopically, all three head and neck cases resembled giant cell angiofibroma/giant cell subtype SFT, and one case showed sheet-like pattern of the multinucleated syncytial cells, creating a biphasic arrangement similar to myofibroma. Both truncal tumor resembled lipomatous type SFT, with one similar to dermatofibrosarcoma protuberans and the other to atypical spindle cell lipomatous tumor. The sella tumor showed morphology of a conventional SFT with high grade sarcomatous transformation. The renal tumor demonstrated a malignant SFT with entrapped benign renal tubules, mimicking a biphase synovial sarcoma or a malignant mixed epithelial and stromal tumor. By immunohistochemistry, all seven SFTs showed diffuse and strong nuclear reactivity to antibody against STAT6. Conclusions Extra-pleural SFTs show a significant heterogeneity of morphology and biological behavior which could cause differential confusion.Careful attention to its characteristic histomorphology with the use of STAT6 immunohistochemistry can help distinguish this tumor from its many mimickers.

Objective To discuss the appropriate timing of providing enteral nutrition through nasojejunal tube for patients with severe craniocerebral injury.Methods 126 cases of patients were divided into 3 groups randomly,providing enteral nutrition through naso-jejunal tube for the first group,the second group and the third group within 12～24 hours,48 hours later and 72 hours later after injury respectively.The nutrition situation of 3 groups was recorded 6 hours later,48 hours later,on the 5th day and the 10th day,including indicators such as total serum protein,blood albumin,serum creatinine,etc and complication cases of diarrhea,hemorrhage of digestive tract,palirrhea,aspiration,inhalation pneumonia and so on within 2 weeks after injury.Results In terms of indicators of albumin,creatinine 48 hours after injury and total protein,albumin and creatinine on the 5th day and 10th day,the first group were better than the second and third group,there were statistic differences between the three groups.Complication comparison within 2 weeks after injury:the difference of palirrhea cases among the three groups was significant,the third group had a higher ratio than the first and second group.And there was no statistic difference in the other indicators like diarrhea,hemorrhage of digestive tract,aspiration and inhalation pneumonia.Conclusions It is high time that patients with simple severe craniocerebral injury are provided with enteral nutrition through naso-jejunal tube within 12 to 24 hours,which can improve patients nutrition situation without the increase of the complications.

Objective To investigate the mRNA expression of UPA and MMP-9 in gastric carcinoma and their correlation with histological type, growth-type, differentiation, vessel invasion, metastasis and prognosis. MethodsIn situ hybridization was used to detect the mRNA expression of UPA and MMP-9. ResultsIn situ hybridization revealed that among 105 cases, the positive rates of UPA mRNA and MMP-9 mRNA were both 58.1%. There was no significant relationship between UPA mRNA and histological type( r _ s = 0.123, P =0.210)and differentiation ( r _ s =0.102, P =0.298)of the tumor. However, there was a significant difference between growth-type( r _ s =0.344, P =0.001),tumor invasion depth( r _ s =0.296, P =0.002),vessel invasion( r _ s =0.198, P =0.042),lymph node( r _ s =0.332, P =0.001)and distant metastasis( r _ s =0.530, P =0.001);there was no significant relationship between MMP-9 mRNA staining and histological type( r _ s =0.143, P =0.145)and differentiation( r _ s =0.102, P =0.298)of the tumor. However, there was a statistically significant difference between growth-type( r _ s =0.267, P =0.006),tumor invasion depth ( r _ s =0.335, P =0.001)、vessel invasion( r _ s =0.209, P =0.032),lymph node( r _ s =0.343, P =0.001)and distant metastasis( r _ s =0.468, P =0.001);There was a positive relationship between UPA mRNA and MMP-9 mRNA expression( r _ s =0.237, P =0.015). The mean survival time in cases with positive UPA mRNA and MMP-9 mRNA expression were significantly shorter than that of cases with negative expression. ConclusionThe mRNA expression of UPA and MMP-9 can predict the invasion and metastasis of gastric carcinoma, They can be used as markers of prognosis of gastric carcinoma in clinical practice.