Objective:To investigate the effects of chronic persistent hypoxia on hepatic function, histological morphology, and ultrastructure in aged mice, and to evaluate the protective role of pyrroloquinoline quinone(PQQ).Methods:Thirty-two 2-month-old (young group)and thirty-two 18-month-old(aged group)male C57BL6/J mice were each randomly divided into four groups (n=8 per group): normoxia+ normal saline (NS)group, normoxia+ PQQ group, hypoxia+ NS group, and hypoxia+ PQQ group.The normoxia+ NS and normoxia+ PQQ groups were housed under normoxic conditions[fraction of inspired oxygen(FiO 2)=21%], while the hypoxia+ NS and hypoxia+ PQQ groups were continuously exposed to a hypoxic environment[FiO 2=(10±0.5)%]simulated by a custom-made hypoxic chamber, maintaining a constant oxygen concentration for 24 hours per day.The normoxia+ NS and hypoxia+ NS groups received daily intragastric administration of NS, whereas the normoxia+ PQQ and hypoxia+ PQQ groups received daily intragastric administration of PQQ disodium salt(8 mg·kg -1·d -1).After 8 weeks of continuous intervention, blood samples were collected to measure red blood cell count, hemoglobin levels, and liver function-related biochemical indicators.Lung tissues were processed for HE staining, and liver tissues were processed for both HE staining and electron microscopy.The histological and ultrastructural features of each group were observed under light and electron microscopy, respectively, and the differences between the groups were compared and analyzed. Results:Compared with the normoxia+ NS groups, both young and aged hypoxia+ NS groups exhibited significant pulmonary arteriole narrowing( P<0.001), with markedly elevated red blood cell count and hemoglobin levels (all P<0.001), which were not alleviated by PQQ.Compared with the young normoxia+ NS group, the young hypoxia+ NS group showed significantly higher alanine aminotransferase (ALT)and lactate dehydrogenase (LDH)levels( Z=2.72, 2.53, P=0.007, 0.011), whereas the young hypoxia+ PQQ group exhibited LDH levels similar to those of the young normoxia+ NS group.The aged hypoxia+ NS group exhibited significant ALT elevation( t=2.66, P=0.013)compared with the aged normoxia+ NS group.Light microscopy revealed hepatocyte ballooning degeneration, mild fatty accumulation, and focal necrosis around central veins in the young hypoxia+ NS group, while the young hypoxia+ PQQ group exhibited no significant pathological damage but displayed numerous deeply stained binucleated hepatocytes.The aged normoxia+ NS group demonstrated hepatocyte ballooning degeneration and inflammatory cell infiltration around central veins, whereas the aged normoxia+ PQQ group exhibited no obvious pathological damage with scattered deeply stained binucleated hepatocytes.The aged hypoxia+ NS group exhibited significant necrosis following physiological oxygen concentration gradient distribution, while the aged hypoxia+ PQQ group displayed no obvious pathological damage with scattered deeply stained binucleated hepatocytes.Electron microscopy revealed that the aged normoxia+ NS group had reduced mitochondrial electron density ( P<0.001)and less developed rough endoplasmic reticulum compared with the young normoxia+ NS group.The young hypoxia+ NS group exhibited a smaller mitochondrial area( P<0.001), decreased mitochondrial matrix electron density( P<0.001), blurred or absent mitochondrial cristae, inactive rough endoplasmic reticulum, and increased accumulation of glycogen and lipid droplets compared with the young normoxia+ NS group, while the young hypoxia+ PQQ group maintained mitochondrial matrix electron density comparable to the young normoxia+ NS group.The aged hypoxia+ NS group exhibited larger mitochondrial area( P=0.001), decreased mitochondrial matrix electron density( P<0.001), blurred or absent mitochondrial cristae, mitochondrial edema, increased lysosomes, and elevated cytoplasmic electron density compared with the aged normoxia+ NS group.The aged hypoxia+ PQQ group exhibited reduced mitochondrial area( P<0.001)and restored mitochondrial matrix electron density to levels comparable with the aged normoxia+ NS group.The aged normoxia+ PQQ group demonstrated increased mitochondrial matrix electron density compared with the aged normoxia+ NS group( P<0.001). Conclusions:Chronic persistent hypoxia induces hepatic functional, histological and ultrastructural damage in mice, with more pronounced effects in aged animals.PQQ provides a certain degree of protection against these injuries.

Objective To investigate the protective effects of Qiji Special Dietary Food on exercise-induced myocardial injury in mice.Methods An exercise-induced myocardial injury model was established using a treadmill running protocol,and at the same time,the modeled mice were administered Qiji Special Dietary Food via oral gavage,followed by a 4-week treadmill exhaustion test.Serum levels of cardiac troponin T(cTnT),creatine kinase(CK),and lactate dehydrogenase(LDH)were measured.Myocardial tissues were analyzed for superoxide dismutase(SOD)activity and malondialdehyde(MDA)content.Histopathological alterations and ultrastructural changes in myocardial tissue were evaluated using light microscopy and transmission electron microscopy(TEM).Results After 4 weeks of exhaustive training,compared to the control group,the model group exhibited significantly elevated serum cTnT,CK,and myocardial MDA levels(P<0.01),along with reduced myocardial SOD activity(P<0.01).Compared to the model group,high-,medium-,and low-dose treatment significantly attenuated the exhaustive exercise-induced increases in serum cTnT level and myocardial MDA content(P<0.01),restoring these indicators to the levels comparable to those of the normal control group(P<0.05).Additionally,all treatment groups had markedly increased myocardial SOD activity,with no significant difference from the normal group.Histopathological and ultrastructural analyses revealed markedly alleviated myocardial damage in the treatment groups,with the medium-dose group exhibiting the most pronounced protective effects.Conclusion Qiji Special Dietary Food demonstrates significant protective effects against exercise-induced myocardial injury in mice,which maybe associated with its antioxidant activity and mitigation of oxidative stress.

Objective:To investigate the effects of chronic persistent hypoxia on hepatic function, histological morphology, and ultrastructure in aged mice, and to evaluate the protective role of pyrroloquinoline quinone(PQQ).Methods:Thirty-two 2-month-old (young group)and thirty-two 18-month-old(aged group)male C57BL6/J mice were each randomly divided into four groups (n=8 per group): normoxia+ normal saline (NS)group, normoxia+ PQQ group, hypoxia+ NS group, and hypoxia+ PQQ group.The normoxia+ NS and normoxia+ PQQ groups were housed under normoxic conditions[fraction of inspired oxygen(FiO 2)=21%], while the hypoxia+ NS and hypoxia+ PQQ groups were continuously exposed to a hypoxic environment[FiO 2=(10±0.5)%]simulated by a custom-made hypoxic chamber, maintaining a constant oxygen concentration for 24 hours per day.The normoxia+ NS and hypoxia+ NS groups received daily intragastric administration of NS, whereas the normoxia+ PQQ and hypoxia+ PQQ groups received daily intragastric administration of PQQ disodium salt(8 mg·kg -1·d -1).After 8 weeks of continuous intervention, blood samples were collected to measure red blood cell count, hemoglobin levels, and liver function-related biochemical indicators.Lung tissues were processed for HE staining, and liver tissues were processed for both HE staining and electron microscopy.The histological and ultrastructural features of each group were observed under light and electron microscopy, respectively, and the differences between the groups were compared and analyzed. Results:Compared with the normoxia+ NS groups, both young and aged hypoxia+ NS groups exhibited significant pulmonary arteriole narrowing( P<0.001), with markedly elevated red blood cell count and hemoglobin levels (all P<0.001), which were not alleviated by PQQ.Compared with the young normoxia+ NS group, the young hypoxia+ NS group showed significantly higher alanine aminotransferase (ALT)and lactate dehydrogenase (LDH)levels( Z=2.72, 2.53, P=0.007, 0.011), whereas the young hypoxia+ PQQ group exhibited LDH levels similar to those of the young normoxia+ NS group.The aged hypoxia+ NS group exhibited significant ALT elevation( t=2.66, P=0.013)compared with the aged normoxia+ NS group.Light microscopy revealed hepatocyte ballooning degeneration, mild fatty accumulation, and focal necrosis around central veins in the young hypoxia+ NS group, while the young hypoxia+ PQQ group exhibited no significant pathological damage but displayed numerous deeply stained binucleated hepatocytes.The aged normoxia+ NS group demonstrated hepatocyte ballooning degeneration and inflammatory cell infiltration around central veins, whereas the aged normoxia+ PQQ group exhibited no obvious pathological damage with scattered deeply stained binucleated hepatocytes.The aged hypoxia+ NS group exhibited significant necrosis following physiological oxygen concentration gradient distribution, while the aged hypoxia+ PQQ group displayed no obvious pathological damage with scattered deeply stained binucleated hepatocytes.Electron microscopy revealed that the aged normoxia+ NS group had reduced mitochondrial electron density ( P<0.001)and less developed rough endoplasmic reticulum compared with the young normoxia+ NS group.The young hypoxia+ NS group exhibited a smaller mitochondrial area( P<0.001), decreased mitochondrial matrix electron density( P<0.001), blurred or absent mitochondrial cristae, inactive rough endoplasmic reticulum, and increased accumulation of glycogen and lipid droplets compared with the young normoxia+ NS group, while the young hypoxia+ PQQ group maintained mitochondrial matrix electron density comparable to the young normoxia+ NS group.The aged hypoxia+ NS group exhibited larger mitochondrial area( P=0.001), decreased mitochondrial matrix electron density( P<0.001), blurred or absent mitochondrial cristae, mitochondrial edema, increased lysosomes, and elevated cytoplasmic electron density compared with the aged normoxia+ NS group.The aged hypoxia+ PQQ group exhibited reduced mitochondrial area( P<0.001)and restored mitochondrial matrix electron density to levels comparable with the aged normoxia+ NS group.The aged normoxia+ PQQ group demonstrated increased mitochondrial matrix electron density compared with the aged normoxia+ NS group( P<0.001). Conclusions:Chronic persistent hypoxia induces hepatic functional, histological and ultrastructural damage in mice, with more pronounced effects in aged animals.PQQ provides a certain degree of protection against these injuries.

Purpose: This study aims to comprehensively evaluate the clinical efficacy of chemotherapy or endocrine therapy maintenance in metastatic breast cancer (MBC) patients. Materials and Methods: The meta-analysis of randomized clinical trials (RCTs) and propensity score matching of multicenter cohort study evaluated MBC patients who underwent first-line chemotherapy or endocrine therapy maintenance. This study is registered with PROSPERO: CRD42017071858 and ClinicalTrials.gov: NCT04258163. Results: A total of 2,867 patients from 15 RCTs and 760 patients from multicenter cohort were included. The results from meta-analysis showed that chemotherapy maintenance improved progression-free survival (PFS) (hazard ratio [HR], 0.63; 95% confidence interval [CI], 0.54 to 0.73; p < 0.001; moderate-quality evidence) and overall survival (OS) (HR, 0.87; 95% CI 0.78 to 0.97; p=0.016; high-quality evidence) than observation. In the cohort study, for hormone receptor–positive MBC patients, chemotherapy maintenance improved PFS (HR, 0.67; 95% CI, 0.52 to 0.85; p < 0.001) and OS (HR, 0.55; 95% CI 0.42 to 0.73; p < 0.001) compared with observation, and endocrine therapy maintenance also improved PFS (HR, 0.65; 95% CI, 0.53 to 0.80; p < 0.001) and OS (HR, 0.55; 95% CI, 0.44 to 0.69; p < 0.001). There were no differences between chemotherapy and endocrine therapy maintenance in PFS and OS (all p > 0.05). Regardless of the continuum or switch maintenance therapy, showed prolonged survival in MBC patients who were response to first-line treatment. Conclusion This study provided evidences for survival benefits of chemotherapy and endocrine therapy maintenance in MBC patients, and there was no difference efficacy between chemotherapy and endocrine therapy maintenance for hormone receptor–positive patients.

Objective To observe the therapeutic effects of Jinsanzhen acupuncture combined with continuous aerobics on the rats with chronic stress-induced depression .Methods The chronic stress-induced depression rat model was developed by using chronic unpredicted mild stimulation ( CUMS) .Thirty male SD rats were randomly divided into the normal control group , the model group ( or the experimental group ) and the treatment group .The rats in the normal control group were given normal feed , but the rats of the model group received stimulation every morning , and the rats of the treatment group also received stimulation every morning , except that they were placed on the treadmill for 30-minute running exercise every afternoon , and then, they were treated with Jinsanzhen acupuncture . Four weeks later , various indicators were detected in the rats of all the groups .Results As compared with that of the rats in the model group, the platform-jumping latency of the rats in the Jinsanzhen therapy combined with continuous aerobics group was elevated , and the number of errors was decreased (P<0.05), open field horizontal and vertical scores were all increased to certain extents (P<0.05), the latency of water maze was decreased and the number of passing through the platform were increased (P<0.05).Pathologic, elec-tron microscopic and Nissl detection results indicated that Jinsanzhen acupuncture combined with continuous aerobics could produce cer -tain effects on the brain tissue of the model group .Conclusion Jinsanzhen acupuncture combined with continuous aerobic exercise could produce good effects on the rats with chronic stress-induced depression .

Objective To research the therapeutic efficacy of the composite gel collagen from marine fish skin on stomatocace in rats.Methods Fifty-six male SD rats were randomly divided into 7 groups:i.e.the 3 dosages groups (the high dosage,medium dosage and low dosage groups),the single gel group,the single collagen group,the positive control group and the model group,each consisting of 8 animals.Thirty-five per cent glacial acetic acid solution was used to develop the rat model of stomatocace,then,changes in stomatocace sizes were observed and detected both before and after medication in all the groups.Changes in the ultrastructure of the ulcer-affected cells in various groups were observed under electronmicroscope and pathological changes in the surface area of ulcer-affected tissues in various groups were further observed by HE staining.Results The composite gel collagen from marine fish skin could reduce ulcer sizes of the affected surface areas in various dosage groups.At day 3 after medication,ulcer sizes in the high dosage,medium dosage and low dosage groups as well as the model group were respectively (8.87 ± 2.11) mm2,(9.50 ± 0.48)mm2,(9.62 ± 1.29) mm2 and (12.31 ± 0.69) mm2.At day 7 after medication,ulcer sizes in the high dosage,medium dosage and low dosage groups as well as the model group were respectively (6.13 ± 2.00) mm2,(7.31 ± 2.70) mm2,(8.92 ± 2.09) mm2 and (10.46 ± 1.33) mm2.At day 10 after medication,ulcer sizes in the high dosage,medium dosage and low dosage groups as well as the model group were respectively(2.52 ± 1.30) mm2,(3.42 ± 1.80) mm2,(4.65 ± 1.51) mm2 and (8.62 ± 1.24) mm2.At day 12 after medication,ulcer sizes in the high dosage,medium dosage and low dosage groups as well as the model group were respectively(0.44 ± 0.92) mm2,(0.41 ± 0.40) mm2,(0.87 ± 1.20) mm2 and (3.12 ± 1.71) mm2.Statistical significance could be noted,when comparisons were made between different dosages groups and the model group (P ＜ 0.05),furthermore,there was a certain dosage-dependent relationship between them.Electronmicroscopy revealed that the tissues of ulcer-affected areas in various dosage groups were generally healed,epithelial cells were tightly lined and degenerative cells could only be occasionally seen between epithelial cells,but without presence of inflammatory cells.Pathological observation revealed that lesion of the tissues in the ulcer-affected areas of various dosage groups was relatively lighter and tissue necrosiswas also relatively lighter.Furthermore,granulation and fibrosis of the affected tissues were quite obvious.Conclusions Composite gel collagen from marine fish skin could produce certain therapeutic efficacy on stomatocace in rats.

Objective To research the therapeutic efficacy of the composite gel collagen from marine fish skin on stomatocace in rats.Methods Fifty-six male SD rats were randomly divided into 7 groups:i.e.the 3 dosages groups (the high dosage,medium dosage and low dosage groups),the single gel group,the single collagen group,the positive control group and the model group,each consisting of 8 animals.Thirty-five per cent glacial acetic acid solution was used to develop the rat model of stomatocace,then,changes in stomatocace sizes were observed and detected both before and after medication in all the groups.Changes in the ultrastructure of the ulcer-affected cells in various groups were observed under electronmicroscope and pathological changes in the surface area of ulcer-affected tissues in various groups were further observed by HE staining.Results The composite gel collagen from marine fish skin could reduce ulcer sizes of the affected surface areas in various dosage groups.At day 3 after medication,ulcer sizes in the high dosage,medium dosage and low dosage groups as well as the model group were respectively (8.87 ± 2.11) mm2,(9.50 ± 0.48)mm2,(9.62 ± 1.29) mm2 and (12.31 ± 0.69) mm2.At day 7 after medication,ulcer sizes in the high dosage,medium dosage and low dosage groups as well as the model group were respectively (6.13 ± 2.00) mm2,(7.31 ± 2.70) mm2,(8.92 ± 2.09) mm2 and (10.46 ± 1.33) mm2.At day 10 after medication,ulcer sizes in the high dosage,medium dosage and low dosage groups as well as the model group were respectively(2.52 ± 1.30) mm2,(3.42 ± 1.80) mm2,(4.65 ± 1.51) mm2 and (8.62 ± 1.24) mm2.At day 12 after medication,ulcer sizes in the high dosage,medium dosage and low dosage groups as well as the model group were respectively(0.44 ± 0.92) mm2,(0.41 ± 0.40) mm2,(0.87 ± 1.20) mm2 and (3.12 ± 1.71) mm2.Statistical significance could be noted,when comparisons were made between different dosages groups and the model group (P ＜ 0.05),furthermore,there was a certain dosage-dependent relationship between them.Electronmicroscopy revealed that the tissues of ulcer-affected areas in various dosage groups were generally healed,epithelial cells were tightly lined and degenerative cells could only be occasionally seen between epithelial cells,but without presence of inflammatory cells.Pathological observation revealed that lesion of the tissues in the ulcer-affected areas of various dosage groups was relatively lighter and tissue necrosiswas also relatively lighter.Furthermore,granulation and fibrosis of the affected tissues were quite obvious.Conclusions Composite gel collagen from marine fish skin could produce certain therapeutic efficacy on stomatocace in rats.

Objective:To compare the adverse reactions of concurrent chemoradiotherapy with docetaxel versus those of sequential and scheduled adjuvant therapy after a modified radical mastectomy in locally advanced breast cancer. Additionally, this work aims to evaluate the safety and feasibility of a synchronous therapy schedule. Methods:A total of 155 female breast cancer patients in the Fourth Affiliated Hospital of Guangxi Medical University were enrol ed from January 2009 to December 2014. Al the patients were diagnosed with infiltrating ductal carcinoma and stage pT3-4, pN1-3c M0, or pAnyTpN2-3cM0 after modified radical mastectomy. After completing the fluorouracil+epirubicin+cyclophosphamide adjuvant chemotherapy, all the patients were randomly divided into two groups by using the method of sealed envelopes. The synchronous group, which received synchronous chemoradiotherapy with docetaxel, comprised 78 cases. The sequential group, which received radiotherapy fol owing docetaxel chemotherapy, comprised 77 cases. The clinical toxic reactions and effects in both groups were assessed after all schedules. Results:A median follow-up period of 39 (16-62) months showed that the radiation side effects of the synchronous and sequential groups were mild. No patients with 3-4 grade radiation-induced skin reactions or symptoms of heart and lung radiation side effects were reported. The rate of 1-2 grade radiation-induced skin reactions was 89.7%(70/78) in the synchronous group and 88.3%(68/77) in the sequential group, but the difference was not statistically significant (P>0.05). The three-year recurrence-free survival rate was 92.3%(72/78) in the synchronous group and 81.8%(63/77) in the sequential group, and the difference was statistically significant (P=0.046). Conclusion:Synchronous chemoradiotherapy with docetaxel as adjuvant therapy exhibited mild and tolerable adverse reactions fol owing modified radical mastectomy in local y advanced breast cancer. Compared with the sequential schedule, the synchronous schedule showed a significantly increased three-year recurrence-free survival rate. Therefore, a synchronous chemo-radiotherapy schedule is safe and feasible and can be used as a treatment option for locally advanced breast cancer.

Objective To establish a single nucleotide polymorphisms genotyping (SNP) method for a convenient, accurate, and routine analysis of clinical samples. Methods Based on the design of oligonucleotide probe, the assay was performed through three steps:the conjunction of the detection probe, universal amplification, labeling and ELISA reaction. The genotype of each SNP was revealed by reading signals of each set of reaction tubes. This assay was applied to detect sixty-two plasma samples of lung cancer for circulating DNA for three SNPs of EGFR, c.2573T＞G(L858R), EGFR, c.2582T＞G＞T(G719C). Results were compared with those obtained by direct sequencing. Results The heterozygote mutation was identified for L858R by both methods, although no mutation was detected for L861Q and G719C. Six samples were identified as heterozygotes with the new method, and only two samples were unambiguously identified as heterozygotes by the direct sequencing. Two additional samples could not be identified as heterozygotes because the peak of mutant allele was very low compared with that of wild allele. Conclusion The developed method enabled accurate identification of SNP in a convenient manner, and which is adapted to routine analysis from heterogeneous samples unambiguously.

Objective To establish a simple,rapid and sensitive nucleotide polymorphisms genotyping method in order to conduct the routine clinical detections under the simple laboratory condition by this method.Methods Based on the ligase-agarose gel electrophoresis,the oligonucleotide detection probes of mutational sites was designed.The detection underwent the detection probe connecting,purification and universal amplification,finally the mutation genotypes of detection sites were judged by the ap-peared bands in the agarose gel electrophoresis(AGE).With the 3 SNP sites EGFR,c.2573T>G(L858R),EGFR,c.2582T> A (L861Q)and EGFR,c.2155 G>T(G719C)in epidermal growth factor receptor(EGFR)gene as the detection objects,the plasmid template and plasma circulating DNA sample in lung cancer were performed the detection.Results The established method was easy to operate with higher specificity and sensitivity.After 20-30 cycles of PCR amplification,the genotype of detection sites was clearly estimated according to the amplification band.When detecting the mixed alleles in the heterogeneous sample,minimal 2.5%mutation alleles could be detected out.This method and the direct sequencing method could respectively detect 6 cases and 2 cases of heterozygotes mutation in the SNP site of L858R among 62 samples of lung cancer.Conclusion The established detection method for SNP genotyping is suitable to the routine mutation detection on the heterogeneous samples under the simple laboratory condi-tion.