Objective To study the feasibility of additive improvement of heart protection by preconditioning with MLA and adenosine.Methods Rabbits of group A were injected with MLA, then 24?h later the hearts were isolated, mounted on a Langendorff apparatus, preconditioned with adenosine, perfused with filtered modified Krebs-Henseleit(K-H) buffer through left atria, instrumented left ventricular function monitor system and paced ventricularly at 180 beat/min. Hearts were arrested with 4?℃ modified St.Thomas solution, stored for 4?h at 4?℃ in this solution, and reperfused with K-H buffer. Left ventricular function, myocardial CK-mb leakage and tissue levels of adenosine triphosphate were measured. Groups B and C were only preconditioned with MLA or adenosine, respectively, while group D was only preserved with the modified St.Thomas solution in 4?℃.Results The left ventricular function recovery as percentage (+dp/dt max) in groups A, B, C was 70.97 ? 17.92 , 65.54 ? 22.62 , 64.36 ? 16.10 , respectively. If compared with group D ( 39.07 ? 13.78 ), the differences were significant ( P

Objective To study the potential improvement of donor heart storage by A1 adenosine receptor-induced delayed preconditioning and the mechanism.Methods Male Wistar rats were randomly divided into 8 groups. Group A was pretreated with 2-chloro-N6-cyclopen-tyladenosine (CCPA),and 24 h later,the hearts were stored in St.Thomas solution for 4 h at 4 ℃ and reperfused with K-H buffer for 1 h,while group B was only injected with vehicle of CCPA. Similarly,group C was administered with CCPA,and 24 h later,subjected to 3 h of hypothermial ischemia and 1 h reperfusion,while groups D,E,F received antisense ODN (AS),sense ODN and scrambled ODN to the initiation site of rat Mn-SOD mRNA before preconditioning with CCPA,respectively. In the meantime,groups G and H were only administered with AS or vehicle of CCPA,respectively. Left ventricular function,myocardial CK-mb leakage,tissue levels of adenosine triphosphate and Mn-SOD were measured.Results The recovery percentage of ?dp/dt max of left ventricle in groups A,C,E and F were much higher than in groups B,D,G and H ( P

Objective PINK1 and Parkin are directly invoveled in the regulation and maintenance of mitochondrial functional morphology. We aim to explore the effect of Wnt2 overexpression on PINK1^B9 Mutant Drosophila and its mechanism in this study. Methods The GAL4-UAS system was used to construct the normal control flies（W1118/ + ；MHC-GAL4/+）, PINK1^B9 transgenic Drosophila model flies（UAS-PINK1^B9 /y；MHC-GAL4 / +；Parkinson's disease model of Drosophila melanogaster), the Wnt2 overexpression flies（UAS-PINK1^B9 /y；MHC-GAL4 / Wnt2 OE） and the Wnt2 RNAi flies（UAS-PINK1^B9 /y；MHC-GAL4 /Wnt2）. On the 5th day, the abnormal wings phenotype rate and flying rate of flies were observed. The contents of Ndufs3 proteins were detected by Western blot. The mRNA expression levels of PGC-1α, Nrf1 and TFAM related to mitochondrial metabolism and synthesis were detected by real-time fluorescence quantitative PCR. The morphology of mitochondria was observed by electron microscopy. Complex I and Complex II function was detected by high-resolution mitochondrial respiratory system. Results Compared with the normal control flies, PINK1^B9 transgenic Drosophila model flies showed increased abnormal wings phenotype rate([1.87±0.06]% vs [68.79±0.70]%), decreased flying rate([ 97.51±0.52)% vs(3.95±0.53)%], and the differences were statistically significant(P<0.05); Compared with PINK1^B9 transgenic Drosophila model flies, the Wnt2 RNAi flies showed decreased abnormal wings phenotype rate[(10.14±1.72)%], increased flying rate([41.83±2.57]%)(P<0.05). Compared with the normal control flies, PINK1^B9 transgenic Drosophila model flies showed decreased expression levels of PGC-1α and Nrf1,Ndufs3 proteins, Complex I and Complex II(P<0.05);On the contrary, the Wnt2 RNAi flies showed increased trends compared with PINK1^B9 transgenic Drosophila model flies(P<0.05). Conclusion Overexpression of Wnt2 protects PINK1^B9 transgenic Drosophila models, which is related to the improvement of mitochondrial function.

Objective:To construct a phage display antibody library of special antigen responding to serum starved U251 cell,from which the serum responding gene and protein would be gotten.Methods:U251 cells were cultured in serum-absent midium for 48 h.Its protein was extracted and used to immunize BALB/c mice.Total RNA of the spleenocytes of immunized mice was extracted.VH and VL were amplified by RT-PCR and were linked into ScFv(Single chain fragment of variation) with a linker.ScFv was recombined to pCANTAB5E vector and was transformed to TG1 strain.Results:The library capacity was up to 3?106 cfu/L.A positive clone was identified from 8 random clones of this library.Conclusion:The special ScFv phage library is constructed successfully.It is the basis for screening special antibodies which can recognize serum responding protein.

ObjectiveTo set up the new lab examination method for 1p, 19q and 10q loss of heterozygosity(LOH) in glioma.MethodsThirty-eight cases of oligodendroglioma were enrolled into the study. Real-time quantitative polymerase chain reaction-based microsatellite analysis was performed on tumor tissues in order to study the status of chromosomes 1p, 19q and 10q.ResultsAmong the 38 cases of oligodendroglioma, 25 cases (65.7%) showed 1p LOH, 26 cases (68.4%) showed 19q LOH, while 5 cases (13.2%) showed 10q LOH.ConclusionReal-time quantitative polymerase chain reaction-based microsatellite analysis is a rapid and specific for detecting LOH in glioma tissues.

Objective:To investigate the application value of near-infrared autofluorescence imaging in identifying and protecting parathyroid glands in endoscopic thyroid surgery. Methods:From May 2022 to February 2023, 158 patients who underwent endoscopic thyroid surgery in the Department of Thyroid and Breast Vascular Surgery of Guilin People's Hospital were selected. The endoscopic fluorescence camera system was used to monitor the parathyroid glands under autofluorescence during endoscopic thyroid surgery. A total of 214 pieces were collected, among which the first 15 cases that could not be preserved in situ during the operation needed to be autotransplanted or the tissue clamped parts that could not be clearly identified as parathyroid glands were sent to fast-frozen pathology to determine whether they were parathyroid glands. Results:Among the first 15 patients who could not be preserved in situ during the operation or whose anatomy could not be clearly defined, 23 parathyroid glands were detected by autofluorescence imaging, 21 parathyroid glands were confirmed by pathology, and 2 were adipose tissue, with an accuracy rate of 91.30%; 158 patients underwent surgery Blood calcium decreased 2 hours after operation compared with preoperative blood calcium（P<0.05）, decreased blood calcium 5 days after operation compared with preoperative blood calcium（P<0.01）, and increased slightly 5 days after the operation compared to blood calcium 2 hours after the operation, but the difference was not statistically significant（P>0.05）; while comparing parathyroid hormone（PTH）, PTH at 2 hours after operation decreased significantly compared with PTH before operation（P<0.01）, and PTH at 5 days after operation compared with PTH before operation PTH also decreased（P<0.01）, but increased compared with PTH 2 hours after operation（P=0.001）. Conclusion:In laparoscopic thyroid surgery, the application of near-infrared autofluorescence imaging technology can help surgeons quickly identify and protect parathyroid glands, and reduce the incidence of permanent hypoparathyroidism. Combining autofluorescence imaging, visual anatomy recognition under magnification of laparoscope, and intraoperative frozen pathological examination "trinity" method can improve the success rate of parathyroid gland recognition.
Humans ; Parathyroid Glands/transplantation* ; Thyroid Gland/surgery* ; Calcium ; Parathyroid Hormone ; Optical Imaging/methods* ; Laparoscopy ; Thyroidectomy/methods*

OBJECTIVE: To investigate the relationship between the severity of allergic asthma and the levels of atrial natriuretic peptide (ANP), and to analyze the potential role of ANP signaling in the pathogenesis of asthma.
 METHODS: We recruited 96 subjects, including 23 healthy volunteers, 25 stable allergic asthmatics, 21 mild allergic asthmatics and 27 moderate allergic asthmatics, from the Affiliated Hospital of Guilin Medical University. ANP, IFN-γ and IL-4 levels in serum were detected by enzyme-linked immunosorbent assay (ELISA), and the mRNA and protein expressions of natriuretic peptide receptor A (NPRA), transcription factor T-bet and GATA3 were measured by RT-PCR and Western blot.
 RESULTS: The levels of ANP in serum and the expressions of NPRA mRNA and protein in the peripheral blood mononuclear cell (PBMC) from the mild asthma group or the moderate group were elevated compared with those in the stable asthma group or the mild group, respectively (P<0.05). Consistently, expressions of GATA3 and levels of IL-4 showed the same tendency (P<0.05). In addition, levels of ANP in serum were positively correlated with the severity of asthma, whereas negatively correlated with the ratio of T-bet/GATA3 and IFN-γ/IL-4 (r=-0.85, P<0.05; r=-0.88, P<0.05, respectively).
 CONCLUSION Levels of ANP signaling in serum were significantly increased with the severity of allergic asthma, suggesting a close relation with the pathogenesis of asthma; ANP signaling may play a role in the pathogenesis of allergic asthma through inducing the Th2-type immune response.
Asthma ; Atrial Natriuretic Factor ; Enzyme-Linked Immunosorbent Assay ; Fetal Proteins ; GATA3 Transcription Factor ; Humans ; Hypersensitivity ; Interleukin-4 ; Leukocytes, Mononuclear ; RNA, Messenger ; Receptors, Atrial Natriuretic Factor ; Signal Transduction ; T-Box Domain Proteins

OBJECTIVE To investigate the effect of bs-5-YHEDA iron chelating peptide combined with semaglutide on the cognitive ability and pathological characteristics of D-Gal-induced Alzheimer's disease(AD) model mice. METHODS Forty mice were randomly divided into 5 groups, namely the healthy control group, PBS group, bs-5-YHEDA iron chelating peptide group, combined treatment group and positive control group, with 8 mice in each group, half of each sex. Except for the healthy control group, D-galactose was injected to induce the AD mice model for 6 weeks. For 3 consecutive weeks starting from the 4th week, the bs-5-YHEDA iron chelating peptide group was injected with bs-5-YHEDA(1 mg·mL–1) once every other day at 200 µL in the tail vein; the bs-5-YHEDA iron chelating peptide(1 mg·mL–1) and semaglutide(25 nmol·kg–1·d–1) were given alternately once a day in the combination treatment group; the positive control group was given memantine(3.3 mg·kg–1·d–1) by gavage every other day. The healthy control group and PBS group were injected with the equal dose of PBS. At the end of treatment, the learning memory ability of mice was detected by the Morris water maze method, whole brain and whole blood were dissected, and pathological changes in hippocampal region were observed by HE staining, and Aβ expression and Tau protein phosphorylation levels were detected by immunohistochemistry, enzyme-linked immunosorbent assay and immunoblotting. RESULTS In the Morris water maze spatial exploration experiment, the differences in the number of times the mice traversed the platform, the ratio of swimming distance to the target quadrant, and the time ratio were statistically significant in each group(P<0.05); compared with the PBS group, the ratio of swimming distance to the target quadrant increased in the combined treatment group, and the differences were statistically significant(P<0.05). The results of HE staining showed that compared with the healthy control mice, the hippocampal area in the PBS group showed reduced levels of pyramidal cells, disorganized arrangement, cell edema, and deep staining of nuclei consolidation. Cellular disorganization, deep staining of nuclei and apoptosis in the hippocampus were significantly improved in each treatment group after drug treatment. Immunohistochemistry and Western blotting results showed that the Aβ expression levels and Tau protein phosphorylation levels were significantly higher in the PBS-administered mice compared with the healthy control mice, and the Aβ expression levels and Tau protein phosphorylation levels were reduced in each group after drug treatment, with statistically significant differences(P<0.01 or P<0.001 ). CONCLUSION The combination of bs-5-YHEDA iron chelating peptide and semaglutide can effectively improve the learning and memory ability and pathological characteristics of AD mice, but from the results of immunohistochemistry and immunoblotting experiments, the improvement of pathological characteristics of AD mice in the combination treatment group is not obvious compared with the single bs-5-YHEDA iron chelating peptide group, suggesting that there may be a threshold effect of our designed dual-target combination treatment on the cognitive improvement of AD mice, and the optimization and validation of the effect of multi-target combination treatment need further study.

Objective To investigate the distribution of aquaporin 4 (AQP4) and inwardly rectifying potassium channel 4.1 (Kir4.1) inthe astrocytes from human mesial temporal lobe epilepsy (MTLE). Methods Hippocampal specimens, including 10 cases of MTLE and 6cases of non-MTLE, were observed under optical and transmission electron microscopy. The distribution of AQP4 and Kir4.1 in astrocyteswas investigated with immunoflurescence. Results Compared with non-MTLE hippocampus, the main structural changes of MTLE includedremarkable hyperplasia astrocytes, serious swelling astrocytes and distinguished astrophy neurons. In non-MTLE hippocampus, immunoflurescencesignals of AQP4 and Kir4.1 were enriched along perivascular astrocyte end-feet domain. However, it reveals significant loss ofAQP4 and Kir4.1 in perivascular astrocyte end-feet domain in MTLE. Conclusion Loss of perivascular AQP4 and Kir4.1 in the humanMTLE may help to understand the roles of astrocyte in MTLE.

OBJECTIVETo investigate the effects of glycosides of tripterygium wilfordii (GTW), methyltestosterone and Zhuanggushenjin capsule on nitric oxide synthase (NOS) in rat testes.

METHODSForty-five rats were equally divided into 5 groups, and respectively given GTW [10 mg/(kg x d)], methyltestosterone [2 mg/(kg x d)], Zhuanggushenjin capsule [0.3 g/(kg x d)], distilled water plus Tween 80 (control I), and distilled water alone (control II) for 4 weeks. At the end of the 5th week, the immunochemical ABC method was used to observe the effects of the three drugs on the NOS positive Leydig cells of the rats.

RESULTSCompared with control II, the GTW group had a significant decrease in the numbers of nNOS and eNOS positive Leydig cells, the methyltestosterone group showed an increase in the number of nNOS but a decrease in that of eNOS positive Leydig cells, and the Zhuanggushenjin group had an increase in the numbers of both nNOS and eNOS positive Leydig cells.

CONCLUSIONGTW can reduce NO production by inhibiting eNOS and nNOS, and hence influence the spermatogenic process. Zhuanggushenjin capsule plays an important role in improving male sexual function by enhancing nNOS and eNOS expression and NO synthesis.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Leydig Cells ; drug effects ; enzymology ; Male ; Methyltestosterone ; pharmacology ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase Type I ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Rats ; Rats, Sprague-Dawley ; Spermatogenesis ; drug effects ; Tripterygium