ObjectiveTo explore the effects of different dosage forms of Kaixinsan （KXS） on the morphology and function of mitochondria in rat models of Alzheimer's disease （AD） and potential mechanisms of action. MethodsMale SD rats were randomly assigned to a sham group， model group， treatment groups receiving KXS decoction， powders， and granules （3.08 g·kg^-1）， as well as donepezil group （0.51×10^-3 g·kg^-1）， with 10 rats in each group. AD model was created using intracerebroventricular injection of streptozocin （STZ）. After 30 days of administration， behavioral assessments were conducted， and mitochondrial morphology was observed using transmission electron microscopy. Mitochondrial respiratory chain complex content was measured via enzyme-linked immunosorbent assay （ELISA）. Changes in mitochondrial membrane potential were measured via JC-1 staining， and superoxide dismutase （SOD） activity and reactive oxygen species （ROS） levels were measured via biochemical assays. The mRNA expression of adenosine 5'-monophosphate-activated protein kinase （AMPK）， peroxisome proliferator-activated receptor gamma coactivator-1α （PGC-1α）， and silent information regulator 3 （SIRT3） was detected by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR）， and Western blot was used to examine the protein expression levels of optic atrophy protein1 （OPA1）， mitochondrial fission protein 1 （FIS1）， AMPK， p-AMPK， PGC-1α， and SIRT3. ResultsCompared with the sham group， rats in the model group had significantly lower recognition index， spontaneous alternation rate， escape latency， number of platform crossings， time spent in the target quadrant， and percentage of distance traveled in the target quadrant distance （P<0.05， P<0.01）. Significant mitochondrial damage was observed in the hippocampal tissue， with a marked decrease in mitochondrial respiratory chain complex content （P<0.01） and reduced mitochondrial membrane potential （P<0.05）. Additionally， the SOD activity was reduced， while ROS levels were elevated （P<0.01）. The mRNA expression of PGC-1α and SIRT3 was significantly downregulated （P<0.01）， along with decreased protein expression levels of OPA1， p-AMPK/AMPK， PGC-1α， and SIRT3， whereas FIS1 protein expression was significantly upregulated （P<0.05， P<0.01）. Compared with the model group， rats in KXS-treated groups （various dosage forms） showed significant improvement in behavioral indexes （P<0.05， P<0.01）， reduced hippocampal mitochondrial damage， and more organized mitochondrial cristae. Mitochondrial respiratory chain complex content was significantly increased （P<0.05， P<0.01）， and mitochondrial membrane potentials were elevated （P<0.05）. SOD activity was elevated， and ROS levels were significantly reduced （P<0.05， P<0.01）. Furthermore， the mRNA expression of PGC-1α and SIRT3 was upregulated， with increased protein levels of OPA1， p-AMPK/AMPK， PGC-1α， and SIRT3， while FIS1 protein expression levels were significantly reduced （P<0.05， P<0.01）. Across the KXS-treated groups， the granule group showed a higher spontaneous alternation rate than the decoction and powder groups （P<0.05）. ConclusionKXS decoction， powders， and granules can improve the learning and memory ability of rats， with granules being the most effective. The mechanism of action may involve activation of the AMPK/PGC-1α/SIRT3 signaling pathway， improvement of the mitochondrial function， and subsequent amelioration of the brain energy metabolism disorders.

ObjectiveTo explore the effects of different dosage forms of Kaixinsan （KXS） on the morphology and function of mitochondria in rat models of Alzheimer's disease （AD） and potential mechanisms of action. MethodsMale SD rats were randomly assigned to a sham group， model group， treatment groups receiving KXS decoction， powders， and granules （3.08 g·kg^-1）， as well as donepezil group （0.51×10^-3 g·kg^-1）， with 10 rats in each group. AD model was created using intracerebroventricular injection of streptozocin （STZ）. After 30 days of administration， behavioral assessments were conducted， and mitochondrial morphology was observed using transmission electron microscopy. Mitochondrial respiratory chain complex content was measured via enzyme-linked immunosorbent assay （ELISA）. Changes in mitochondrial membrane potential were measured via JC-1 staining， and superoxide dismutase （SOD） activity and reactive oxygen species （ROS） levels were measured via biochemical assays. The mRNA expression of adenosine 5'-monophosphate-activated protein kinase （AMPK）， peroxisome proliferator-activated receptor gamma coactivator-1α （PGC-1α）， and silent information regulator 3 （SIRT3） was detected by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR）， and Western blot was used to examine the protein expression levels of optic atrophy protein1 （OPA1）， mitochondrial fission protein 1 （FIS1）， AMPK， p-AMPK， PGC-1α， and SIRT3. ResultsCompared with the sham group， rats in the model group had significantly lower recognition index， spontaneous alternation rate， escape latency， number of platform crossings， time spent in the target quadrant， and percentage of distance traveled in the target quadrant distance （P<0.05， P<0.01）. Significant mitochondrial damage was observed in the hippocampal tissue， with a marked decrease in mitochondrial respiratory chain complex content （P<0.01） and reduced mitochondrial membrane potential （P<0.05）. Additionally， the SOD activity was reduced， while ROS levels were elevated （P<0.01）. The mRNA expression of PGC-1α and SIRT3 was significantly downregulated （P<0.01）， along with decreased protein expression levels of OPA1， p-AMPK/AMPK， PGC-1α， and SIRT3， whereas FIS1 protein expression was significantly upregulated （P<0.05， P<0.01）. Compared with the model group， rats in KXS-treated groups （various dosage forms） showed significant improvement in behavioral indexes （P<0.05， P<0.01）， reduced hippocampal mitochondrial damage， and more organized mitochondrial cristae. Mitochondrial respiratory chain complex content was significantly increased （P<0.05， P<0.01）， and mitochondrial membrane potentials were elevated （P<0.05）. SOD activity was elevated， and ROS levels were significantly reduced （P<0.05， P<0.01）. Furthermore， the mRNA expression of PGC-1α and SIRT3 was upregulated， with increased protein levels of OPA1， p-AMPK/AMPK， PGC-1α， and SIRT3， while FIS1 protein expression levels were significantly reduced （P<0.05， P<0.01）. Across the KXS-treated groups， the granule group showed a higher spontaneous alternation rate than the decoction and powder groups （P<0.05）. ConclusionKXS decoction， powders， and granules can improve the learning and memory ability of rats， with granules being the most effective. The mechanism of action may involve activation of the AMPK/PGC-1α/SIRT3 signaling pathway， improvement of the mitochondrial function， and subsequent amelioration of the brain energy metabolism disorders.

The guideline and item classification approach represents a distinct historical method ology, first introduced by ZHU Xi in his Compendium of Comprehensive Mirror for Aid in Government during the Southern Song Dynasty. LOU Ying later adapted this framework to the field of traditional Chinese medicine, critically analyzing and addressing the limitations of prior academic theories. By drawing upon the rich intellectual traditions of classical texts and historical works, LOU Ying restructured and refined his ideas, ultimately establishing the "guideline and item" medical theory. This innovative framework profoundly influenced LOU Ying′s work, shaping both the compilation of medical texts and the development of novel theoretical approaches in traditional Chinese medicine. In disease differentiation and treatment, LOU Ying proposed the theory of guideline and item classification, that is, taking yin and yang as two guidelines, taking qi and blood, exterior and interior, upper and lower parts, deficiency and excess, cold and heat as ten items, and constructing a syndrome differentiation system focusing on qi and blood, focusing solely on upper and lower parts, distinguishing exterior and interior, carefully examining deficiency and excess, and distinguishing cold and heat. The guideline and item classification system integrates and harmonizes multiple syndrome differentiation method ologies, including the eight-principle pattern identification, viscera syndrome differentiation, differentiation of syndromes according to the pathogenesis of Sanjiao, and qi-blood-fluid pattern identification. As a precursor to modern method of syndrome differentiation and treatment, it possesses unique theoretical significance and considerable clinical relevance. Its applicability to diagnosing and treating various diseases, coupled with its greater operational practicality compared to other methods, highlights its value in contemporary clinical practice. This system offers a complementary and enhanced framework for syndrome differentiation and treatment, warranting further adoption and promotion in the clinical setting.

Objective　To analyze the clinical distribution of carbapenem-resistant Enterobacter cloacae (CR-ECL) and changes in carbapenemase enzyme types in Ningbo and to provide a guide for the rational application of antibiotics in clinical practice. Methods　Clinical isolates of CR-ECL were selected in a hospital in Ningbo from 2012 to 2022. Bacterial identification and drug sensitivity testing were performed using the VITEK-2 COMPACT automated microbiology system. Strain confirmation was conducted using the VITEK MS mass spectrometry system. The presence of CR-ECL was confirmed using the E-Test method. Carbapenemase production in CR-ECL was determined using a rapid carbapenemase detection kit and polymerase chain reaction (PCR). Results　A total of 1 428 isolates of ECL were detected in a hospital in Ningbo from 2012 to 2022, of which 36 strains were CR-ECL, accounting for 2.52%. The top three sample sources of CR-ECL were sputum (52.78%), urine (19.44%), and bile (13.89%). The patients were mainly from neurosurgery wards (22.22%), general surgery (13.89%), and ICU wards (11.11%). The separation rate of CR-ECL showed no significant difference between different genders and age groups. The resistance rates of CR-ECL to ertapenem, imipenem, and meropenem were 94.44%, 58.33%, and 36.11% respectively. The resistance rates of CR-ECL to third/fourth generation cephalosporins, aztreonam, enzyme-inhibitor complexes, and quinolones were all above 69.50%, significantly higher than those of non-carbapenem-resistant Enterobacter cloacae (NCR-ECL) strains (P<0.05). For aminoglycosides, CR-ECL and NCR-ECL both maintained low resistance rates (0% and 0.22%, respectively). Among the 36 CR-ECL strains, 31 strains (86.11%) were carbapenemase producers, including three types: NDM (28 strains, 77.78%), KPC (4 strains, 11.11%), and IMP (2 strains, 5.56%). The other two carbapenemase types, OXA-48 and VIM, were not detected. The consistency rate between the rapid detection of carbapenem enzyme type and PCR detection was 100%. Conclusions　The detection rate of CR-ECL in a hospital in Ningbo showed a trend of fluctuation. CR-ECL strains are distributed across different departments and various samples of hospitals, with no differences in distribution observed between genders or age groups. Clinical isolated CR-ECL mainly produced NDM-type carbapenemases, but the enzyme types such as KPC and IMP have also emerged in recent years. The rapid identification of carbapenemase types can play an important guiding role in the clinical treatment of CR-ECL infection.

This research project was designed to explore the molecular basis of the loss of contact inhibition in hepatoma cells by regulating the cell growth density of hepatic cells (L02) and hepatoma cells (HepG2) respectively. Analyzing the character of morphology, the change of cytoskeleton, the ability of deformation, the expression and distribution of E-cadherins of hepatic cells and hepatoma cells with different density, we found: Hepatoma cells' E-cadherins increased when compared to the hepatic cells'; Hepatic cells' up-regulated E-cadherins, and with their increased growth density, hepatic cells gathered in the contacted areas; Hepatoma cells, however, tended to down-regulate the expression of E-cadherins, and they kept the fusion distribution. The migration rate and net migration distance of these two kinds of cells were inhibited by growth density. Hepatoma cells kept the strong ability of migration, but the migration trace discretization of hepatic cell decreased with the increase of growth density. Hepatoma cells kept the high discretization of migration trace in different growth density. These different results show that hepatic cells are in positive correlation with E-cadherins distribution, and are in negative correlation with its migration. There is no aggregation tendency seen with respect to hepatoma cells' E-cadherins. The effect of hepatoma cells' growth density on migration is not obvious.
Cadherins ; metabolism ; Cell Count ; Cell Line ; Cell Movement ; Hep G2 Cells ; Humans ; Liver ; cytology ; metabolism

Objective To explore the influence of integrin redistribution on hepatoma cell alignment and migration and the influence of cytoskeleton reassembly on integrin redistribution by the method of mechanical loading unloading and fibronection(FN) coating. Method By using immuneofluescence staining, cofocal laser scanning microscopy and quantitative morphological analysis, integrin distribution change and crtoskeleton assembly adjustment were observed and the deformation of cell movement was tested and analyzed quantitatively. Results (1) cells with different forms have different integrin expressions and distribution features. The β1 integrin expression for spreading cells was higher than that for round (nonspreading) cells. For spreading cells, the strongest staining was found towards the attachment surface. While for round (nonspreading) cells, the integrin staining on the free surface towards medium was stronger than that towards the attachment surface. (2) After 5 hours of mechanical stretch, the β1 integrin expression for both spreading and round cells increased, and distribution peaks towards the attachment surface broadened. At 1 hour after unloading, the β1 integrin expression decreased and the distribution of integrin staining showed the tendency of dispersion, especially for round cells. (3) After coating the substrates with FN, the β1 integrin expression increased. The integrin staining for either spreading or round cells was more towards the attachment surface to reduce the migration of hepatoma cells. 4) After 5 hours of mechanical stretch, 60% of cells showed their orientation of major axes distributed between 70°～110° towards the stretching direction, and the cytoskeleton aligned vertically to the stretching direction. Cytoskeletons were found significantly depolymerized at 1 hour of unloading. Conclusions The change of integrin distribution is affected by cytoskeleton aligned and the number of ligand. The distribution feature of the whole integrin expression on the surface of individual round cells is related to their stronger invasion and metastasis capability.