ObjectiveTo observe the effect of Qishao capsules on renal injury in db/db mice with diabetic kidney disease （DKD），and explore its mechanism of protecting the kidney by inhibiting podocyte apoptosis. Methodsdb/m mice （7 mice） were used as the normal group，and db/db mice （35 mice） were randomly divided into a model group，a dapagliflozin group （0.001 g·kg^-1·d^-1），and low-，medium-，and high-dose groups of Qishao capsules （0.341 3，0.682 5，and 1.365 g·kg^-1·d^-1，respectively）. Drug intervention lasted for 8 consecutive weeks. After sampling，the serum renal function indicators ［creatinine（SCr），and urea nitrogen（BUN）］，fasting blood glucose （FBG），24 h urinary protein quantification （24 h-UTP）， and other indicators of the mice were measured. The pathological tissue morphology of the kidney was observed by periodic acid-silver methenamine （PASM） and Masson's trichrome （Masson） staining. Immunohistochemical detection of cysteine-dependent aspartate-specific protease （Caspase）-3 and B-cell lymphoma 2 （Bcl-2） was performed. Western blot was used to detect the protein expression of Caspase-8，Caspase-7，Caspase-3， and other molecules. Terminal deoxynucleotidyl transferase dUTP nick End labeling （TUNEL） staining was used to observe apoptosis in renal tissue. Immunofluorescence staining of Wilms tumor suppressor gene-1 （WT-1） was used to observe podocyte injury. Real-time polymerase chain reaction （Real-time PCR） was employed to detect Caspase-8 and Caspase-3 mRNA expression in the kidneys of experimental mice. Data analysis was conducted using statistical software GraphPad Prism 10. ResultsCompared with the normal group，the model group showed significantly increased 24 h-UTP，SCr，BUN，and FBG （P<0.01）. Additionally， PASM staining of renal tissue showed increased glycogen deposition，glomerular hypertrophy，and thickening of the basement membrane. Masson staining showed collagen fiber proliferation in renal tissue. Transmission electron microscopy showed thickening of the renal tissue basement membrane and fusion or loss of foot processes in the model group. The immunohistochemical results showed that Caspase-3 in the kidneys of the mice in the model group significantly increased （P<0.01），while the Bcl-2 protein expression level decreased significantly （P<0.01）. The number of TUNEL positive cells in renal tissue significantly increased （P<0.01）. The positive expression of WT-1 in podocytes was significantly reduced （P<0.01）. Western blot analysis showed significant upregulation of Caspase-8，Caspase-7，and Caspase-3 protein expression in renal tissue （P<0.01）. The mRNA expression levels of Caspase-8 and Caspase-3 in the kidneys increased significantly （P<0.01）. Compared with the model group，the Qishao capsules groups can significantly reduce the levels of 24 h-UTP，SCr，BUN，and FBG （P<0.01）. The pathological damage of renal tissue and podocyte injury were improved to some extent. Immunohistochemistry in the kidney showed a significant decrease in Caspase-3 （P<0.01） and a significant increase in Bcl-2 protein expression level （P<0.01）. Additionally， there were a significant decrease in the number of TUNEL positive cells in renal tissue （P<0.01），a significant increase in WT-1 positive expression in podocytes （P<0.01），marked downregulation of Caspase-8，Caspase-7，and Caspase-3 protein expression in renal tissue （P<0.05，P<0.01），and a significant decrease in Caspase-8 and Caspase-3 mRNA expression levels （P<0.01）. ConclusionQishao capsules can inhibit podocyte apoptosis by regulating the expression of Caspase-8，Caspase-7， and Caspase-3，thereby improving the diabetic renal injury in db/db mice.

Objective:To observe the effects of Qizhi Tongluo Prescription on renal interstitial fibrosis in rats with membranous nephropathy based on the Shh/Gli1 signaling pathway; To explore its intervention mechanism.Methods:Totally 60 male SD rats were divided into blank group ( n=10) and model group ( n=50) using random number table method. The model of membranous nephropathy was established according to the modified Border method. The successfully modeling rats were divided into model group, benazepril group and Qizhi Tongluo Prescription low-, medium- and high-dosage groups using random number table method. Benazepril group was gavaged with benazepril hydrochloride 10 mg/kg, Qizhi Tongluo Prescription low-, medium- and high-dosage groups were gavaged with Qizhi Tongluo Prescription solution 1.22 g/kg, 2.43 g/kg and 4.86 g/kg, and blank group and model group were gavaged with equal volume of normal saline, once a day, for 4 weeks. The 24-hour urine was collected to detect the 24-hour urinary protein quantification, and the blood was taken from the abdominal aorta to detect the levels of total cholesterol (TC), triglyceride(TG), and serum albumin(ALB); the pathological changes of rat kidney were observed by light microscope and transmission electron microscope; the protein expressions of sonic hedgehog factor (Shh), zinc finger protein 1 (Gli1) and α-smooth muscle actin (SMA) in renal tissues were detected by immunohistochemistry; the protein expressions of Shh, Gli1, α-SMA, TGF-β1, Collagen Ⅳ and plasminogen activator inhibitor-1 (PAl-1) in renal tissues were detected by Western blot. Results:Compared with the model group, the quantitative level of 24-hour urinary protein of rats in each administration group decreased ( P<0.05), serum TC and TG levels increased ( P<0.05), ALB level decreased ( P<0.05), the positive expressions of Shh, Gli1, α-SMA protein in renal tissue decreased ( P<0.05), and the protein expressions of Shh, Gli1, α-SMA, Collagen Ⅳ, TGF-β1, PAI-1 in renal tissue decreased ( P<0.05). Conclusion:Qizhi Tongluo Prescription can improve renal interstitial fibrosis in membranous nephropathy rats, possibly by inhibiting the Shh/Gli1 signaling pathway to delay renal interstitial fibrosis.

Objective To investigate the clinicopathological characteristics of colorectal cancer patients with different mismatch repair (MMR) statuses and their correlation with KRAS/NRAF/BRAF (KNB) gene mutations. Methods The clinicopathological data of 477 patients with colorectal cancer were collected, and MMR, microsatellite instability (MSI), and KNB status were detected via immunohistochemistry (IHC), PCR–capillary electrophoresis, and next-generation sequencing (NGS), respectively. The clinicopathological features of patients with different MMR statuses and correlations with KNB mutations were analyzed. Results Compared with the patients in the pMMR group, the patients in the classical dMMR group were younger, included more females, and exhibited more tumors in the right colon, mostly mucinous adenocarcinoma and poorly differentiated tumors (all P<0.05). The tumors in the nonclassical dMMR group were commonly found in the right colon and were prone to special histologic types (all P<0.05). MLH1-PMS2 codeletion, BRAF mutation, and KRAS G13 codon mutation were common in patients in both the classical and the nonclassical dMMR groups (both P<0.05). The results of MMR IHC (100%) were highly consistent with those of MSI PCR (99.1%). Patients in the classical dMMR group with KRAS mutations were younger, included more males, and were prone to specific histologic types, but distant metastasis was rare (all P<0.05). Conversely, lymph node metastasis was rare in patients in the nonclassical dMMR group with KRAS mutations (P=0.005). The mutation rate of the MSH6 gene was relatively high in the nonclassical dMMR group (P=0.002), and all patients presented complete deletion of MLH1-PMS2 combined with nonclassical expression of MSH6 (100%). Five patients with medullary carcinoma components had complete deletions of MLH1-PMS2. Among the five patients, three had combined nonclassical expression of MSH2/MSH6. Two of the three patients carried the MSH6 gene c.3261 locus mutation. Conclusion The clinicopathologic features of patients with classical/nonclassical dMMR colorectal cancer differ from those of patients with pMMR, and MMR IHC could be used to predict effectively the MSI status. The clinicopathologic features differ between classical and nonclassical dMMR colorectal cancer patients with KRAS mutations, but both groups present codeletion of MLH1-PMS2, BRAF mutation, and KRAS G13 codon mutation. In patients with nonclassical dMMR, complete deletion of MLH1-PMS2 combined with nonclassical expression of MSH6 is common. Mutations in the MSH6 gene may play a key role in the development of colorectal cancer with medullary carcinoma components.

ObjectiveTo observe the effect of salvianolate on the protein expressions of adenosine monophosphate （AMP）-activated protein kinase （AMPK）， silent information regulator 1 （SIRT1） and peroxisome proliferator-activated receptor-γ coactivator-1α （PGC-1α）， autophagy and apoptosis in kidney tissue of rats with membranous nephropathy （MN）， and to explore its possible molecular mechanism against MN. MethodEighty male SD rats were randomly divided into normal group， model group， benazepril hydrochloride group （10 mg·kg^-1）， and salvianolate low-， medium-， and high-dose groups （16.7， 33.3 and 66.7 mg·kg^-1）. The rats were modeled by injection of cationized bovine serum albumin （C-BSA） into the tail vein. After successful modeling， rats in the administration groups were given corresponding doses of drugs for 4 consecutive weeks， and then 24-hour urine， serum and kidney tissue were collected for the detection of 24-hour urinary protein （UTP）， blood urea nitrogen （BUN）， serum creatinine （SCr）， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， C reactive protein （CRP）， glutathione peroxidase （GSH-Px）， superoxide dismutase （SOD）， and malondialdehyde （MDA）. The pathological changes of kidneys were observed by light microscope， electron microscope and immunofluorescence. Western blot was used to detect the protein expressions of phospho-AMPK （p-AMPK）， AMPK， phospho-SIRT1 （p-SIRT1）， SIRT1 and PGC-1α in rat kidney tissue. The protein expressions of autophagy-specific gene （Beclin-1）， microtubule-associated protein 1 light chain 3 （LC3） Ⅱ， ubiquitin-binding protein （p62）， B cell lymphoma （Bcl-2）， Bcl-2-associated X （Bax）， and cysteine aspartic protease-7 （Caspase-7） in rat kidney tissue were determined by immunohistochemistry （IHC）. ResultCompared with the conditions in the normal group， the levels of UTP， IL-6， TNF-α， CRP and MDA in the model group were increased （P<0.05） while the levels of SOD and GSH-Px were decreased （P<0.05）， and there was no difference in BUN and SCr. Compared with the model group， the administration groups had lowered UTP， IL-6， TNF-α， CRP and MDA （P<0.05） while elevated SOD and GSH-Px （P<0.05）. It could be seen from hematoxylin and eosin （HE） staining， Masson staining， immunofluorescence and electron microscopy that the pathological damage of rat kidney tissue in the model group was significant， but after treatment with benazepril hydrochloride and salvianolate， the pathological damage of kidney cells was gradually improved. The expressions of p-AMPK/AMPK， p-SIRT1/SIRT1， PGC-1α， Bcl-2， Beclin-1 and LC3Ⅱ in rat kidney in the model group were lower than those in the normal group （P<0.05） while the expressions of Bax， Caspase-7 and p62 were higher （P<0.05）. Compared with the model group， benazepril hydrochloride group and salvianolate groups had an up-regulation in the expressions of p-AMPK/AMPK， p-SIRT1/SIRT1， PGC-1α， Bcl-2， Beclin-1 and LC3Ⅱ in the kidney （P<0.05） while a down-regulation in the expressions of Bax， Caspase-7 and p62 （P<0.05）. ConclusionThe protective effect of salvianolate on the kidneys of MN rats may be related to the activation of AMPK/SIRT1/PGC-1α signaling pathway， the up-regulation of autophagy and the reduction of apoptosis.

ObjectiveTo observe the effect of Dahuang Xiezhuo prescription on the clinical symptoms， blood uric acid， and renal tubular function of patients with immunoglobulin A （IgA） nephropathy in stages 1-2 of chronic kidney disease （CKD） complicated with hyperuricemia （HUA）. MethodSixty patients with IgA nephropathy in stages 1-2 of CKD complicated with HUA of spleen and kidney deficiency and combined turbidity and blood stasis syndromes were randomly divided into an observation group and a control group， with 30 cases in each group. The patients in the control group received basic treatment， i.e.， losartan potassium tablets 50-100 mg/time， once per day， and sodium bicarbonate tablets 0.5 g/time， three times per day by oral administration， combined with low-salt， low-fat， and low-purine diet. The patients in the observation group received Dahuang Xiezhuo prescription on the basis of basic treatment， one dose per day， twice a day in the morning and evening with warm water. Both groups were treated for two months. The total scores of traditional Chinese medicine（TCM）syndrome， blood pressure， 24 h urinary protein （24 h UTP）， blood urea nitrogen （BUN）， serum creatinine （SCr） ［glomerular filtration rate （eGFR） was calculated by CKD-epidemiology collaboration （CKD-EPI） formula］， serum uric acid （SUA）， and renal tubular function indexes ［urinary α₁-microglobulin （α₁-MG）， urinary β₂-microglobulin （β₂-MG）， urinary kidney injury molecule-1 （KIM-1）， and neutrophil gelatinase-associated lipocalin （NGAL）］ of the two groups before treatment and two months after treatment were recorded. The clinical efficacy of the two groups was evaluated two months after treatment. ResultAfter 2 months of treatment，the total effective rate in the observation group was 81.48%（22/27），higher than 50.00%（14/28） in the control group（χ² =6.661，P<0.05）. The total scores of TCM syndrome， 24 h UTP， and SUA in the observation group and the observation group were lower than those before treatment （P<0.05）， and compared with the control group after treatment， the observation group decreased more significantly （P<0.05）. After treatment， the blood pressure in the observation group and the observation group was lower than that before treatment （P<0.05）， and there was no significant difference in blood pressure between the two groups after treatment. After treatment， the levels of urinary α₁-MG， β₂-MG， KIM-1， and NGAL in the two groups were lower than those before treatment （P<0.05）， and the observation group was lower than the control group after treatment （P<0.05）. There were no significant inter-group and intra-group differences in BUN， SCr， and eGFR levels before and after treatment. There were no obvious abnormalities in blood routine， liver function， and electrolytes before and after treatment in the two groups， and no adverse reactions such as allergies occurred. ConclusionDahuang Xiezhuo prescription can effectively improve the clinical symptoms of IgA nephropathy with HUA （CKD1-2） patients with spleen and kidney deficiency and combined turbidity and blood stasis syndromes， reduce blood uric acid level， alleviate renal tubular injury， and protect the kidney. The curative effect is better than that of basic treatment.

ObjectiveTo observe the intervention effect of Dahuang Xiezhuo prescription （DHXZ） on inflammation and suppressor of cytokine signaling 3 （SOCS3）/Toll-like receptor 4 （TLR4） pathway in rats with chronic renal failure （CRF）， and to explore its molecular mechanism in alleviating renal inflammatory response. MethodThe 90 male SD rats， 15 were randomly selected as sham group， and the remaining 75 were used as modeling group to replicate CRF rat model by 5/6 nephrectomy. After successful modeling， the rats were randomly divided into model group， DHXZ low-， medium-， high-dose groups （6.825， 13.65， 27.3 g·kg^-1） and Niaoduqing Granules group （2.6 g·kg^-1）. The drug intervention groups received corresponding drugs by gavage for 8 consecutive weeks. After administration， hematoxylin-eosin （HE） staining and Masson staining were used to observe the morphological changes of rat renal tissue， and blood creatinine （SCr）， blood urea nitrogen （BUN） and blood uric acid （UA） were tested. Enzyme-linked immunosorbent assay （ELISA） was performed to detect the serum contents of interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α） and C-reactive protein （CRP）. The mRNA expressions of SOCS3 and TLR4 in renal tissue were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， and the protein expressions of SOCS3， TLR4， nuclear transcription factor （NF-κB） and myeloid differentiation factor （MyD88） were detected by Western blot. Immunohistochemistry was used to determine the protein expressions of NF-κB， MyD88， NOD-like receptor protein 3 （NLRP3） and melanoma deficiency factor 2 （AIM2）. ResultCompared with the sham group， the model group had a significant inflammatory response in renal tissue， and an increase in blood SCr， BUN， UTP， IL-6， TNF-α and CRP （P<0.05）. The protein and mRNA expressions of SOCS3 in renal tissue of rats in the model group were lower while the protein expressions of TLR4， NF-κB， MyD88， NLRP3 and AIM2 and the mRNA expression of TLR4 were higher than those in the sham group （P<0.05）. Compared with the model group， DHXZ and Niaoduqing granules groups presented markedly reduced inflammatory response in renal tissue and decreased blood SCr， BUN， UTP， IL-6， TNF-α and CRP （P<0.05）. Additionally， DHXZ and Niaoduqing granules up-regulated the protein and mRNA expressions of SOCS3 in renal tissue while down-regulated the protein expressions of TLR4， NF-κB， MyD88， NLRP3 and AIM2 and the mRNA expression of TLR4 （P<0.05）. ConclusionDHXZ can reduce the release and expression of inflammatory factors， inhibit the inflammatory response and improve renal function， and the mechanism may be related to the regulation of SOCS3/TLR4 signaling pathway.

ObjectiveTo observe the effect of Dahuang Xiezhuo prescription on the changes in renal pathology and reactive oxygen species （ROS）/thioredoxin-interacting protein （TXNIP）/NOD-like receptor protein 3 （NLRP3） pathway expression in the kidney tissues of rats with 5/6 nephrectomy， and to explore the mechanism of Dahuang Xiezhuo prescription in protecting renal function and delaying renal interstitial fibrosis and the possibility. MethodNinety healthy male SD rats were randomly divided into a sham operation group， a model group， low， medium， and high-dose （6.825， 13.65， 27.30 g·kg^-1） Dahuang Xiezhuo prescription groups， and a Niaoduqing granule group （2.60 g·kg^-1）. Except the sham operation group， 5/6 nephrectomy was used to replicate the rat model of chronic renal failure （CRF）. After modeling， each administration group was given the corresponding dose of drug suspension by intragastric administration， once a day for consecutive 8 weeks. After administration， serum creatinine （SCr） and urea nitrogen （BUN） levels and 24 h urinary protein quantification （UTP） levels were detected. Western blot assay was used to detect the protein expressions of thioredoxin （TRX）， TXNIP， and NLRP3. The protein expressions of TRX， TXNIP， NLRP3， apoptosis-associated speck-like protein containing a CARD （ASC）， transformation growth factor-β （TGF-β）， Collagen Ⅳ， α-smooth muscle actin （α-SMA）， and fibronectin （FN） were detected by immunohistochemistry. ResultAs compared with the sham operation group， serum levels of SCr， BUN， and UTP in the model group were increased （P<0.05）， TRX， TXNIP， NLRP3， ASC， TGF-β， Collagen Ⅳ， α-SMA， and FN proteins were increased （P<0.01）， and renal interstitial fibrosis significantly occurred. As compared with the model group， the levels of SCr， 24 h BUN， and UTP in the low， medium， and high-dose Dahuang Xiezhuo prescription groups and the Niaoduqing granule group were decreased to varying degrees （P<0.05）， TRX， TXNIP， NLRP3， ASC， TGF-β， Collagen Ⅳ， α-SMA， and FN were decreased （P<0.01）， and renal interstitial fibrosis was improved to varying degrees. ConclusionDahuang Xiezhuo prescription can protect renal function and delay renal interstitial fibrosis in rats with CRF.

China/epidemiology* ; Diabetes Mellitus, Type 2/etiology* ; Dyslipidemias/epidemiology* ; Humans ; Incidence ; Prospective Studies

Objective To evaluate the left ventricular diastolic function of patients with normal left ventricular ejection fraction ( LVEF) by echocardiography and real‐time cardiac catheter measurement ,and improve the accuracy and reliability of echocardiographic diagnosis . Methods One hundred and twenty patients with know n or suspected coronary artery disease w ho underwent coronary angiography and left ventricular catheterization were prospectively selected from July 2017 to January 2018 in the Affiliated Hospital of Jiangsu University . According to the left ventricular end diastolic pressure ( LVEDP) real‐time measurement ,the patients were divided into groups of LVEDP ≤15 mm Hg ( 43 cases ) and LVEDP ＞ 15 mm Hg ( 77 cases) . General data were compared and the difference of echocardiographic parameters between the two groups were analyzed ,and the ROC curve of each echocardiographic parameter for diagnosing LVEDP was draw n . Results T he parameters including flow propagation velocity ( VP) ,the ratio of filling fraction of E and A ( E/A) ,early diastolic filling deceleration time ( DT ) ,the duration of mitral A ( A‐dur ,) mitral annulus velocity at the septal side ( e′sep) ,systolic pulmonary venous flow velocity ( PVs) ,diastolic pulmonary venous flow velocity ( PVd ) and PVs/PVd were used to the diagnosis of the increasing of LVEDP ,however their accuracies were low ( AUC between 0 .5～0 .7) . T he parameters including left atrial volume index ( LAVI ) , tricuspid regurgitation ( T Rmax ) ,mitral annulus velocity in lateral wall of left ventricle ( e′lat ) ,average e′,E/e′sep ,E/e′lat ,average E/e′,velocity of pulmonary vein atrial reversal ( PVa) ,pulmonary vein atrial reversal duration ( Pva‐dur) ,the difference between the duration of pulmonary venous A wave and mitral A wave( PvaD‐AD) were also used to the diagnosis of the increasing of LVEDP , but their accuracies were still poor ( AUC between 0 .7～0 .9 ) . According to the real‐time left ventricular pressure measurement and different parameters of echocardiography ,the multivariate regression equation :LVEDP＝ 0 .292 LAVI + 0 .35 PVa + 0 .04 T Rmax + 0 .075 ( PvaD‐AD ) －0 .109 PVs －6 .773 was put forward as a correction standard ,the accuracy of the diagnosis of LVEDP was significantly improved ( AUC ＝0 .922) . Conclusions T he assessment of left ventricular diastolic function needs to be performed comprehensively with multiple parameters . T he multiple regression equation can accurately evaluate left ventricular diastolic function in patients with normal LVEF .