Objective The special features of discoid meniscus(DM) symptoms， the relationship between the symptoms and the signs of DM, the type and scale of DM injury as well as the prognosis were studied. Methods According to the analysis of 64 cases of DM diagnosed by arthroscopy, a comparitive analysis was made and the following topics were addressed: the distribution of the complete type of DM in patients of different age groups, the signs of locking and non-locking, with or without effusion, snapping and non-snapping, a comparison of the distribution of the snapping in different groups of injury types, a comparison of the distribution of the flexion contracture in different types of injury, a comparison of the distribution of the rates of the total and subtotal menisectomy in different groups divided according to the period of pain, the presence of effusion, the presence of locking, snapping, quadriceps atrophy and extension function. The data was analysed statistically. Results There is significant difference in the distribution of the complete type DM between 20 and over 20-years of age(P0.05). Conclusion The complete type of DM was mostly seen in cases below 20 years of age and was rarely seen in cases with the locking sign. Snapping sign was mostly seen in cases of complicated and extensive injuries and was one of the indications of arthroscopic surgery. Knee flexion contracture was mostly seen in cases of longitudinal and extensive injuries. The estimation of the results of arthroscopic surgery for DM is difficult on the bases of the clinical symptoms and signs.

Objective To investigate the expressions of advanced glycation end products (AGEs) and their receptors in keloid. Methods Serum and skin tissue specimens were collected from 20 patients with keloid, 20 patients with hyperplastic scar and 20 normal human controls. Fluorospectrophotometer was used to measure the serum level of AGEs, and immunohistochemistry and Western blotting to detect the expressions of AGEs and AGER in skin tissue specimens. Results The serum level of AGEs was (0.713 ± 0.098) AU/ml and (0.699 ± 0.077) AU/ml respectively in patients with keloid and those with hypertrophic scar, significantly higher than that in normal controls (0.179 ± 0.056 AU/ml, F = 283.82, P < 0.01 ). A positive expression of AGEs and AGER was observed in tissue specimens of keloid and hyperplastic scar, but not in the control specimens. As Western blotting showed, the expressions of AGEs and AGER were higher in tissue specimens of keloid and hyperplastic scar than in the control specimens (F = 18.04, 42.80, both P < 0.05), while no significant difference between keloid and hyperplastic scar tissue specimens was observed (P> 0.05). Conclusion There is a high expression of AGEs and AGER in keloid, which may contribute to the development of keloid.

Objective To investigate the expressions of advanced glycosylation end products (AGE) in skin of mice with diabetes mellitus (DM) for different durations,and to evaluate their influence on collagen fibers.Methods Forty healthy 8-week-old male C57BL/6J mice were divided into DM group (n =20) and control group (n =20) to receive multiple intraperitoneal injections of low dose streptozotocin (50 mg/kg) and citric acid buffer (0.1 mol/L),respectively,for 5 consecutive days.Ten mice were sacrificed in each group on week 4 and 12 respectively after the last intraperitoneal injection,and full-thickness skin tissue samples were harvested from the middorsal region of each mouse.Then,hematoxylin-eosin (HE) staining was performed to observe histological changes,and total collagen content was estimated according to hydroxyproline content measured by an alkalinehydrolysis method.The cross-linking degree of collagen was determined by Edman degradation method using pepsin,the mRNA expression level of collagen type Ⅰ and Ⅲ by real-time quantitative PCR,the content of AGE by fluorospectrophotometry and Western blotting,and the level of malondialdehyde (MDA) by using a thiobarbituric acid method.Statistical analysis was carried out by t test.Results As light microscopy showed,the skin became obviously thinner in the diabetic mice with a progressive decrease in the number of collagen fibers in comparison with the control mice.On week 4 and 12 after the last injection,the diabetic mice exhibited a significant reduction in the content of hydroxyproline ((684.5 ± 76.7) vs.(787.7 ± 87.7) rg/g,(558.1 ± 73.1) vs.(757.8 ± 75.3) mg/g,both P ＜ 0.01) and in the levels of cross-linked collagen as well as mRNA expressions of collagen Ⅰ and Ⅲ (P ＜ 0.01 or 0.05),but a significant increase in the content of AGE ((37.47 ± 10.65) vs.(26.39 ± 3.74) AUF/mg hydroxyproline,(47.70 ± 5.66) vs.(29.91 ± 6.50) AUF/mg hydroxyproline,both P ＜ 0.01) and MDA ((6.62 ± 0.47) vs.(4.82 ± 0.56) μmol/L,(8.63 ± 0.36) vs.(5.15 ± 0.46) μmol/L,both P＜ 0.01) in skin tissue,compared with the control mice.The level of non-cross-linked collagen in skin tissue was also lower in the diabetic mice than in the control mice on week 12 (P ＜ 0.05).Moreover,the contents of hydroxyproline and the expression levels of collagen I in skin were significantly lower (P ＜ 0.05),but the levels of AGE and MDA were significantly higher (P ＜ 0.01) in the diabetic mice on week 12 than in those on week 4.Conclusions The characteristics of collagen fibers in skin are altered in diabetic mice when compared with normal control mice,which may be associated with increased AGE content and oxidative injury in skin.

AIM:To examine the expression profiles of both genes and proteins in hippocampus of rats with temporal lobe epilepsy(TLE)for revealing the molecular mechanisms of TLE and looking for the candidate targets and new therapeutic approaches in clinical practice.METHODS:Rat temporal lobe epilepsy was induced by administration of lithium chloride and pilocarpine(LiCl-PILO).The expression spectra of genes and proteins were constructed through the techniques of cDNA microarray,two-dimensional(2D)electrophoresis and Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS).Subsequently,the differentially expressed genes and proteins were identified and analyzed.RESULTS:There were 192 genes of differential expression observed in hippocampal tissues of LiCl-PILO-induced temporal lobe epilepsy,and 159 genes have been registered in Genbank database,in which 84 genes were up-regulated while 75 genes were down-regulated.78 protein spots of differential display were screened out,in which 31 proteins were detected to be down-regulated and 47 were up-regulated.Finally,5 proteins were identified.CONCLUSION:These genes and proteins found in our study may play pivotal roles in the pathogenic mechanisms of epilepsy and may promise new therapeutic targets for refractory epilepsy in the future.

Objective:To study the protective effect of neoeriocitrin on PC12 cell injury induced by amyloid β protein fragment 25-35 (Aβ 25-35), and explore its role in preventing and treating AD. Methods:The MTT method was used to detect the effect of Neoeriocitrin on the proliferation of normal PC12 cells and Aβ 25-35 damaged PC12 cells, and the intervention concentration of neoeriocitrin solution was screened. The PC12 cells were divided into the normal control group, model group, estradiol group and neoeriocitrin group according to the random number table method. The normal control group and model group were cultured with DMEM for 24 h; DMEM and 1×10 -3 μmol/L estradiol solution were added to the estradiol group; DMEM and 6×10 4 μmol/L neoeriocitrin solution were added to the neoeriocitrin group. After 2 hours of culture, except for the normal control group, the remaining groups were added with 20 μmol/L Aβ 25-35 stock solution, and the culture was continued for 22 h. AnnexinV-FITC/PI double labeling was used to detect apoptosis rate, Western blot method was used to detect estrogen receptor β (estrogen receptor β, ERβ) and p-P38/P38 protein expression. The content of acetylcholine (ACh) was detected, and the activity of Choline acetyl transferase (ChAT) and Acetylcholin esterase (AChE) in the supernatant of PC12 cells were detected. Results:Compared with the model group, the cell apoptosis rate [(8.080 ± 0.578)% vs. (18.500 ± 0.870)%] significantly decreased ( P<0.01), the expression of ERβ protein (0.348 ± 0.042 vs. 0.273 ± 0.006) significantly increased ( P<0.01), p-P38/P38 protein expression (0.372 ± 0.058 vs. 0.571 ± 0.063) significantly decreased ( P<0.01), the content of ACh [(14.319 ± 1.039) μg/mg vs. (9.157 ± 1.605) μg/mg], ChAT activity [(0.715 ± 0.053) U/mg vs. (0.280 ± 0.093) U/mg] significantly increased ( P<0.01), AChE activity [(2.607 ± 2.048) U/mg vs. (6.038 ± 1.867) U/mg] significantly reduced ( P<0.01). Conclusions:Neoeriocitrin can promote the proliferation of PC12 cells damaged by Aβ25-35, inhibit cell apoptosis, and has a certain cytoprotective effect. Its mechanism may be related to the regulation of ERβ expression and inhibition of P38 protein phosphorylation, thereby improving the function of the cholinergic system related.

Objective:To observe the effect of warming needle therapy combined with Tuina(Chinese therapeutic massage)on the clinical symptoms and anxiety state of patients with cervical vertigo accompanied by anxiety. Methods:Seventy patients with cervical vertigo accompanied by anxiety state were divided into an observation group and a control group using the random number table method,with 35 patients in each group.In addition to disease education,the observation group was treated with warming needle therapy and Tuina 3 times a week,and the control group was treated with betahistine mesylate tablets orally 3 times a day.Both groups were treated for 4 weeks.The changes in the scores of the evaluation scale for cervical vertigo(ESCV),self-rating anxiety scale(SAS),and Hamilton anxiety rating scale(HAMA)were compared between the two groups after treatment. Results:One case in the observation group dropped out due to failure to cooperate with the treatment during the study.In the treatment group,the total effective rate and the cured plus markedly effective rate were 94.1%and 50.0%,respectively,and 88.6%and 8.6%in the control group,respectively;the differences between the two groups were statistically significant(P<0.05).After treatment,the ESCV,SAS,and HAMA scores of both groups decreased significantly compared with those before treatment(P<0.01),and the scores of the observation group were lower than those of the control group(P<0.01 or P<0.05). Conclusion:Warming needle therapy combined with Tuina can significantly improve the clinical symptoms of patients with cervical vertigo accompanied by anxiety state.

OBJECTIVETo label embryonic stem (ES) cells with enhanced green fluorescent protein (EGFP) on the hypoxanthineguanine phosphoribosyl transferase (HPRT) gene locus for the first time to provide a convenient and efficient way for cell tracking and manipulation in the studies of transplantation and stem cell therapy.

METHODSHomologous fragments were obtained by polymerase chain reaction (PCR), from which the gene targeting vector pHPRT-EGFP was constructed. The linearized vector was introduced into ES cells by electroporation. The G418(r)6TG(r) cell clones were obtained after selection with G418 and 6TG media. The integration patterns of these resistant cell clones were identified with Southern blotting.

RESULTSEGFP expressing ES cells on the locus of HPRT were successfully generated. They have normal properties, such as karyotype, viability and differentiation ability. The green fluorescence of EGFP expressing cells was maintained in propagation of the ES cells for more than 30 passages and in differentiated cells. Cultured in suspension, the "green" ES cells aggregated and formed embryoid bodies, retaining the green fluorescence at varying developmental stages. The "green" embryoid bodies could expand and differentiate into various types of cells, exhibiting ubiquitous green fluorescence.

CONCLUSIONSThis generation of "green" targeted ES cells is described in an efficient protocol for obtaining the homologous fragments by PCR. Introducing the marker gene in the genome of ES cells, we should be able to manipulate them in vitro and use them as vehicles in cell-replacement therapy as well as for other biomedical and research purposes.

Animals ; Cells, Cultured ; Chromosome Mapping ; Embryo, Mammalian ; cytology ; Green Fluorescent Proteins ; Hypoxanthine Phosphoribosyltransferase ; genetics ; Luminescent Proteins ; metabolism ; Mice ; Recombination, Genetic ; Stem Cells ; metabolism ; Transgenes

OBJECTIVETo evaluate the effect of vascular endothelia1 growth factor (VEGF) on the hematogenous metastasis of non-small cell lung cancer (NSCLC).

METHODSThe identification of lung cancer cells in the peripheral blood were carried out by cytological, immunohistocytologica1 and immunofluorecent stains respectively, following isolation of cytokeratin-expressing cells with magnetic activated cell sorting. The quantification of cancer cells in the blood was performed according to the established flow cytometric assay. The plasma VEGF was measured by commercially available ELISA kit.

RESULTSThe lung cancer cells in the blood, showing a remarkable nuclear polymorphism, expressed the epithelial marker cytokeratin and telomerase reverse transcriptase (hTERT). These cells were stained positive by an NSCLC-specific monoclonal antibody S5Al0-2, but negative by antibodies against CD34 and CD45 antigens. Using the flow cytometric assay, 44 cases (28.6%) of l54 NSCLC patients were found to have cancer cells in their blood, with the incidence of positive cases correlated with the stage of disease. The plasma VEGF level was significantly increased in NSCLC patients in comparison with healthy individuals and patients with benign pulmonary diseases. This level was correlated with the stages of disease in patients with adenocarcinoma. In patiens with cancer cells in their blood, a higher level of plasma VEGF was related with an increased number of cancer cells.

CONCLUSIONThe plasma VEGF level is increased in NSCLC patients with approximate1y one fourth to have cancer cells in the peripheral blood. In these patients, increased VEGF level promotes hematogenous tumor metastasis, as indicated by a much higher number of cancer cells in the blood.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung ; blood ; pathology ; DNA-Binding Proteins ; Endothelial Growth Factors ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunohistochemistry ; Keratins ; analysis ; Lung Neoplasms ; blood ; pathology ; Lymphokines ; blood ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplastic Cells, Circulating ; chemistry ; pathology ; Telomerase ; analysis ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

Objective To investigate the effect of comprehensive intervention led by nursing staffers on the drug compliance in patients with hepatitis B virus-related hepatocellular carcinoma (HCC) after receiving transcatheter arterial chemoembolization (TACE) treatment.Methods By using random sampling method,a total of 96 patients with hepatitis B virus-associated HCC,who had been treated with TACE and had taken nucleotide analogue drug for one month in the interventional department of a certain grade Ⅲ tumor hospital,were enrolled in this study.Under the premise of informed consent,comprehensive intervention,which was led by nursing staffers and was participated by both doctors and nurses,was conducted.Results After comprehensive intervention,the average score of drug compliance was (93.670±6.046) points,while the pre-intervention average score of drug compliance was (82.040±10.024) points,the difference between the two was statistically significant (P＜0.05).The ratio of patients who showed good drug compliance changed from pre-intervention 62.4％ to post-prevention 97.8％(P＜0.05).The post-prevention of patients,whose hepatitis B virus deoxyribonucleic acid (HBVDNA) level was within the normal range,was 45.16％,which was highcr than the pre-intervention ratio of 15.05％,the difference between the two was statistically significant (P＜0.05).Conclusion The comprehensive intervention led by nursing staffers can effectively improve the drug compliance in patients with hepatitis B virus-associated HCC after receiving TACE,and can reliably control serum HBVDNA level as well.

Objective:To compare the clinical efficacies between arthroscopic anatomic single bundle reconstruction of posterior cruciate ligament (PCL) assisted by internal tension relieving technique combined with rapid rehabilitation and anatomic single bundle reconstruction combined with conventional rehabilitation in the treatment of PCL rupture.Methods:A retrospective cohort study was used to analyze the clinical data of 88 patients with PCL rupture admitted to First Affiliated Hospital of Kunming Medical University from September 2016 to September 2020. The patients included 65 males and 23 females, aged 18-55 years [(39.3±10.8)years]. Forty-four patients underwent arthroscopic anatomic single bundle reconstruction of PCL assisted by internal tension relieving technique combined with rapid rehabilitation therapy (tension-relieving group), and 44 patients underwent arthroscopic routine anatomic single bundle reconstruction of PCL combined with conventional rehabilitation (traditional group). The two groups were compared before and at 3, 12, and 24 months after surgery regarding the following items: International Knee Literature Committee (IKDC) score, Hospital for Special Surgery (HSS) score, Lysholm score, knee motion cycle (maximum stride length, minimum stride length, and stride frequency) and 6 kinematic indicators (angle of forward and back extension, angle of internal and external rotation, angle of internal and external rotation, up and down displacement, internal and external displacement, and forward and back displacement). The Marburger arthroscopy score (MAS) and Professor Ao Yingfang′s improved score by secondary arthroscopy were compared between the two groups at 12 months after surgery. The perioperative complications were observed.Results:All patients were followed for 24-36 months [(25.5±6.3)months]. In tension-relieving group and the traditional group, the values of IKDC score were (71.8±9.8)points and (68.5±6.5)points at 3 months after surgery, (87.6±6.0)points and (87.6±5.5)points at 12 months after surgery, and (95.5±3.1)points and (92.8±11.6)points at 24 months after surgery, respectively. The values were gradually increased, significantly higher than those before surgery [(48.1±16.9)points and (47.1±15.0)points] (all P<0.05). There were no significant differences between the two groups at each time point (all P>0.05). In tension-relieving group and the traditional group, the values of HSS score were (74.2±6.2)points and (68.4±9.5)points at 3 months after surgery, (91.9±5.4)points and (88.4±4.7)points at 12 months after surgery, and (97.1±2.0)points and (96.2±2.8)points at 24 months after surgery, respectively. The values of HSS score gradually increased, significantly higher than those before surgery [(57.5±17.7)points and (56.8±14.3)points] (all P<0.05). At 3 and 12 months after surgery, the values of HSS score in the tension-relieving group were significantly higher than those in the traditional group (all P<0.05), but others were not significantly different between the two groups (all P>0.05). In tension-relieving group and the traditional group, the values of Lysholm score were (74.2±14.9)points and (70.3±7.5)points at 3 months after surgery, (90.9±6.1)points and (88.7±4.7)points at 12 months after surgery, and (96.9±3.0)points and (96.3±2.8)points at 24 months after surgery, respectively. The values of Lysholm score were gradually increased, significantly higher than those before operation [(48.7±20.7)points and (48.2±19.9)points] (all P<0.05). There were no significant differences between the two groups at any time points (all P>0.05). At 3, 12, and 24 months after surgery, the motion cycle (maximum stride length, minimum stride length and stride frequency) and 6 kinematic indicators (angle of forward bending and backward extension, angle of internal and external rotation, angle of internal and external rotation, internal and external displacement, up and down displacement, and forward and backward displacement) of knee joint were significantly improved in both groups compared with those before surgery (all P<0.05). At 3, 12, and 24 months after surgery, the forward and backward displacement in the tension-relieving group was significantly decreased than that in the traditional group (all P<0.05), but others were not significantly different between the two groups (all P>0.05). The MAS was rated as excellent to good in 14 patients and fair to poor in 2, with the excellent and good rate of 87.5% (14/16) in the tension-relieving group, while the score was rated as excellent to good in 11 patients and fair to poor in 3, with the excellent and good rate of 78.6% (11/14) in the traditional group ( P>0.05). The Professor Ao Yingfang′s improved score was (10.6±1.5)points in the tension-relieving group, markedly higher than that in the traditional group [(9.6±2.3)points] ( P<0.05). No perioperative complications were observed. Conclusion:Compared with anatomic single bundle reconstruction combined with conventional rehabilitation, arthroscopic anatomic single bundle reconstruction of PCL assisted by internal tension relieving technique combined with rapid rehabilitation have higher subjective function score, better anteroposteric stability during knee movement, and better results of secondary microscopy.