Objective To investigate the expression of MMP-9 genes in primary hepatocellular carcinoma(HCC).Methods RT-PCR was used to detect MMP-9 mRNA in surgically extracted specimens from 28 patients with HCC.Results The expression of MMP-9 mRNA was significantly higher in tumor than in corresponding nontumoral tissue(P0.05).Meanwhile,the expression of MMP-9 mRNA in HCC was significantly correlated to the existence of portal vein tumor thrombus(P

Abstract: Objective To observe the curative effect of thread-hanging combined with cotton plug on stage Ⅲ paronychia. Methods Sixty-one patients with stage Ⅲ paronychia were selected and randomly divided into a treatment group and a control group. The treatment group (n=31) was treated with thread-hanging and tampon under local infiltration anesthesia, and changed dressing and tampon every day after operation. After the wound healed, the patient soaked his feet in warm water every day and changed the tampon himself until the symptoms subsided, and the knot did not receive special treatment, and the nail plate would naturally shed as it outgrew the paronychia. The control group (n=30) was treated with thread-hanging and nail groove reconstruction under nerve block anesthesia, and the dressing was changed every day after operation. After thread removal, the patients soaked their feet in warm water every day until the symptoms subsided, and the knot was not specially treated, and it naturally fell off with the growth of the deck beyond the nail groove. The postoperative Visual Analog Scale (VAS) pain score, pain duration, wound healing time, cure rate, effective rate and recurrence rate of paronychia, and patients' satisfaction with the operation were compared between the two groups. Results Compared with the control group, the treatment group had lower VAS pain scores on the first and third postoperative days (2.1±0.3) and (0.2±0.1) vs. (6.3±0.1) and (3.2±0.2), respectively, shorter duration of pain and wound healing time (3.3±0.3) days and (10.1±0.5) days vs. (5.2±0.3) days and (15.2±0.3) days, respectively, higher cure rate (87.1% vs. 66.7%), lower failure rate (12.9% vs. 33.3%), lower recurrence rate (7.4% vs. 20.0%), and higher patient satisfaction (97.0% vs.75.3%). The treatment group showed significant superiority over the control group in all outcomes. Conclusion For patients with stage Ⅲ paronychia, thread-hanging combined with cotton tampon without nail groove reconstruction is advantageous as it avoids additional skin trauma, and does not affect the nail appearance and normal periungual barrier after healing, , reduces patient discomfort, and shortens the time off work, resulting in a higher cure rate. This treatment approach is therefore worth promoting in clinical practice.

BACKGROUND:At present, the separation and culture technique of chondrocytes has been mature, but the chondrocytes grow slowly which are prone to degenerate using the present technique. It is not conducive to the fol ow-up test.
OBJECTIVE:To investigate and improve the separation and culture method of articular chondrocytes of New Zealand rats at 4 weeks of age.
METHODS:New Zealand rats aged 4 weeks were selected to take cartilage tissues from the bilateral knees that were resected under aseptic condition. Chondrocytes were isolated by type Ⅱ col agenase enzyme digestion and mechanical isolation method. The cells were cultured and passaged, and then identified by morphologic observation, toluidine blue staining and type Ⅱ col agen enzyme immunohistochemical methods. Growth curve was pictured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method.
RESULTS AND CONCLUSION:Inverted microscope observation showed that the primary cultured chondrocytes adhered at 6 hours after cultivation. The monolayer formation occurred at 72 hours after cultivation, and the cells were ready to be passaged at 96 hours after cultivation. In the fourth generation, some cells represented a spindle-like appearance. In the fifth generation, most cells turned into irregular shape appearance, and cellproliferation capacity diminished. Toluidine blue staining showed that the nuclei of cultured chondrocytes were blue and cytoplasm was pale blue. Immunofluorescent staining showed that cultured chondrocytes had a positive expression of col agen type Ⅱ and the color was tawny. Proliferative rate of chondrocytes in the first to third generations had no differences (P<0.05), while differences were found compared with the fourth generation in 4-7 days (P<0.05) and the fifth generation in 1-7 days (P<0.05). The results indicate that type Ⅱ col agenase enzyme digestion and mechanical isolation method is successful for isolating, cultivating New Zealand rat articular chondrocytes in vitro, and the first to third generations can be the best choice for the experiments of knee osteoarthritis.

Objective To investigate the expression of aquaporin 3 (AQP3) in lesions of four cuta-neous tumors. Methods Immtmohistochemistry was utilized to measure the expression of AQP3 in tissue samples from 30 patients with seborrheic keratosis, 15 patients with Bowen's disease, 32 patients with squa-mous cell carcinoma, 17 patients with malignant melanoma and 15 normal human controls. Results AQP3 was observed in all the tissue samples from both patients and controls. A significant increment was noticed in the expression of AQP3 in patients with Bowen's disease, squamous cell carcinoma and malignant melanoma compared with the normal controls (all P < 0.01), while no significant difference was found between patients with seborrheic keratosis and the controls (P > 0.05). The highest expression of AQP3 was obtained in lesions from patients with squamous cell carcinoma and malignant melanoma, followed by those with Bowen's disease (both P < 0.01), and there was no significant difference between squamous cell carcinoma and malignant melanoma (P > 0.05). The differentiation status of squamous cell carcinoma significantly corre-lated with the expression of AQP3 (P < 0.01). AQP3 was significantly higher in malignant melanoma with metastasis than that in malignant melanoma without metastasis (P < 0.05). Conclusion The expression of AQP3 is upregulated in malignant skin tumors.

AIM:To investigate the proliferation property of stable chemerin gene knockdown vascular smooth muscle cells ( VSMCs) and to explore its mechanism .METHODS:The normal VSMCs , chemerin gene interfering control VSMCs and stable chemerin gene knockdown VSMCs were divided into normal group , PDGF group, control group and knockdown group .The VSMCs in PDGF group were given platelet-derived growth factor-BB ( PDGF-BB) to initiate proli-feration.The cell counting and BrdU assay were employed to investigate the proliferation property of VSMCs .The mitogen-activated protein kinase ( MAPK) signal pathway was determined by Western blot .RESULTS:The cell number and BrdU A value in PDGF-BB-treated VSMCs significantly increased as compared with the normal VSMCs ( P<0.05 ) .Compared with the normal VSMCs , the chemerin knockdown VSMCs showed obviously decreased cell number and BrdU A value ( P<0.05).Simultaneously, no significant difference in the proliferation of VSMCs between the normal VSMCs and the control VSMCs was observed.No significant difference of the protein levels of p-ERK1/2, ERK1/2, p-p38 MAPK and p38 MAPK among 4 kinds of VSMCs was found .The protein level of p-JNK in PDGF-BB-treated VSMCs was up-regulated, while it was down-regulated in chemerin knockdown VSMCs compared with the normal VSMCs .CONCLUSION: Chemerin pro-motes the proliferation of mouse vascular smooth muscle cells by up-regulating p-JNK production .

Objective To investigate the natural infection of Theiler's murine encephalomyelitis virus (TMEV) in mice,and to survey the distribution of virus in tissues and the changes of serum antibody in the experimentally TMEV-infected mice.Methods Enzyme linked immunosorbent assay (ELISA) and fluorescence quantitative RT-PCR (qRT-PCR) assay were used to detect the antibody and nucleic acid of TMEV in clinical samples.These samples included SPF mice collected from Guangdong area in 2010-2015,mice obtained from a non-barrier laboratory rodent colony,and wild Rattus norvegicus live-trapped around the non-barrier laboratory rodent colony.36 ICR mice were intracerebrally inoculated with TMEV BeAn strain.The clinical signs of the animals were observed daily post-inoculation.Three mice were euthanatized at day 0,3,7,10,17,21,31,39 and 46 post-inoculation (dpi),respectively.Tissue and serum samples were collected for TMEV detection.Results The TMEV antibody and nucleic acid positive rates of SPF mice collected from Guangdong area in 2010-2015 were 5.29％ (n=2834) and 27.27％ (n=457),respectively.The TMEV antibody and nucleic acid positive rates of the mice obtained from a non-barrier laboratory rodent colony were 71.95％ (n=82) and 53.66％ (n=82),respectively.The TMEV nucleic acid positive rate of wild Rattus norvegicus was 25.93％ (n=27).In the TMEV positive mice,only two mice showed obvious clinical symptoms.The cecal contents,feces and brain samples were the best candidates for qRT-PCR assay.The viral nucleic acid could be detected in the brain,heart,liver,lung and stomach of ICR mice at 3 dpi,but no viral nucleic acid was detected in the spleen,kidney,and cecum.The viruses in liver,heart,lungs and stomach were completely cleared at 10 dpi,and the viruses persisted in the brain throughout the experiment.The TMEV antibody could be detected at 7 dpi,and then the antibody positive rate reached 100％ at 17 dpi.The antibody level increased gradually and maintained up to 46 days.ICR mice showed latent infection after TMEV inoculation,with no obvious symptoms and eye pathological changes.Conclusions The experimental mice and wild Rattus norvegicus in Guangdong area are both infected with TMEV,and the infection rate is high.The mice inoculated with TMEV BeAn strain show latent infection.The TMEV antibody produced in mice can be detected at 7 dpi and persisted until the end of the experiment.The viruses are found in the liver,heart,lung and stomach for a short time,but are persisted in the brain for a long time.There is a good consistency of TMEV detection between qRT-PCR and ELISA.The qRT-PCR assay can be used as a powerful complement method for the national standard of laboratory animals.

Objective:To evaluate the efficacy and safety of small incision surgery combined with multi-point skin fixation in the treatment of axillary osmidrosis.Methods:104 patients with axillary osmidrosis who were treated in the dermatology department of the Third Hospital of Changsha from January 2017 to December 2020 were retrospectively analyzed. They were divided into the observation group (56 cases) and the control group (48 cases). Both groups were treated with small incision pruning combined with porous drainage. On this basis, the observation group was treated with multi-point skin fixation gauze compression bandage, while the control group was treated with conventional gauze stacking compression bandage. The efficacy, satisfaction, postoperative wound healing time and complication rate of the two groups were compared.Results:The effective rate of the observation group and the control group were 96.43%(54/56) and 95.83%(46/48) respectively, with no significant difference ( P>0.05). Compared with preoperative, the Visual Analogue Scale (VAS) score of patients in the two groups was significantly lower after operation, and the difference was statistically significant (both P<0.05). The satisfaction of patients in the observation group was higher than that in the control group [(4.05±1.15)points vs (3.19±1.00)points], and the difference was statistically significant ( t=4.10, P<0.05). The wound healing time in the observation group was shorter than that in the control group, and the incidence of complications was lower than that in the control group, with statistically significant difference (all P<0.05). Conclusions:Small incision surgery combined with multi-point skin fixation for the treatment of axillary osmidrosis has good curative effect, short postoperative wound healing time and fewer complications, and improved patient satisfaction, which can be popularized in clinical application.

Objectives: . We aimed to develop a new calculation model for calcium requirements in dialysis patients following parathyroidectomy. Methods: . A total of 98 patients with secondary hyperparathyroidism receiving parathyroidectomy from January 2014 to January 2022 were enrolled in this study. Among these patients, 78 were randomly selected for construction of the calcium requirement calculation model, and the remaining 20 patients were selected for model validation. The calcium requirement model estimated the total calcium supplementation for 1 week after surgery using variables with significant relationships in the derivation group by stepwise multiple linear regression analysis. Bias, precision, and accuracy were measured in the validation group to determine the performance of the model. Results: . The model was as follows: calcium requirement for 1 week after surgery=33.798–8.929×immediate postoperative calcium+0.190×C-reactive protein–0.125×age+0.002×preoperative intact parathyroid hormone+0.003×preoperative alkaline phosphatase (R2=0.8). The model was successfully validated. Conclusion . We generated a novel model to guide calcium supplementation. This model can assist in stabilizing the serum calcium levels of patients during the early postoperative period. Furthermore, it contributes to the individualized and precise treatment of hypocalcemia in patients following parathyroidectomy.

Objective To establish a rapid,specific and sensitive TaqMan real-time fluorescence quantitative PCR assay for detection of murine polyomavirus ( MPyV) .Methods The specific primers and TaqMan probe were designed based on genome sequence of MPyV.The primers amplified a 69 bp fragment.After optimizing the reaction system and reaction condition, the standard curve was plotted by detecting recombinant plasmid standards.The specificity, sensitivity and reproducibility of this method were evaluated.In addition, samples of lungs, spleens and feces obtained from experimentally infected mice and 86 clinical samples were used to validate the efficacy of this real-time PCR assay.Results The specificity assay showed that this assay could specifically detect MPyV and the sensitivity for MPyV was about 100 copies/well.The coefficients of variation ( CV) of both intra-assay and inter-assay were less than 1.13%.All of the samples from experimentally infected mice were positive for MPyV and 3 out of 86 clinical samples were positive by this TaqMan-PCR detection with a positive rate of 3.5%.Conclusions The real-time fluorescence quantitative TaqMan-PCR assay established in this study has high specificity, sensitivity and stability.It can be used for clinical diagnosis, routine detection and epidemiological investigation of murine polyomavirus infections.

Objective:To investigate the clinical characteristics of the disease spectrum of pediatric hyper blood immunoglobulin E (IgE).Methods:A total of 498 children with total serum IgE ≥ 5×10 5 IU/L admitted to the Department of Pediatrics, Sun Yat-Sen Memorial Hospital of Sun Yat-Sen University from January 2012 to December 2018 were enrolled.Their clinical data, etiology distribution, clinical manifestations, laboratory results, treatment and outcomes were retrospectively analyzed.According to serum total IgE level, patients were divided into mildly increased IgE group (5×10 5-<10×10 5 IU/L), moderately increased group (10×10 5-<20×10 5 IU/L), and severely increased group (≥20×10 5 IU/L). The distribution of disease types among the 3 groups were compared. Results:(1) Allergic disease (213 cases) was the most common etiology in children with hyper blood IgE, and infectious disease (163 cases), mycoplasma pneumoniae (109 cases) and EB virus (120 cases) were common pathogens.(2) The incidence of allergic diseases (45.0%) and infectious diseases (42.2%) in the mildly increased group was significantly higher than that in the moderately increased group (40.8%, 26.2%, respectively) and the severely increased group(38.9%, 12.2%, respectively) (all P<0.001). The incidence of immune diseases(18.5%), tumors and hematological diseases (5.4%) in the moderately increased group was significantly higher than that of the mildly increased group (4.4%, 2.0%, respectively) (all P<0.001). The incidence of immune diseases (34.4%), tumors and hematological diseases (11.1%) in the severely increased group was significantly higher than that of the mildly increased group(4.4%, 2.0%, respectively) and the moderately increased group (18.5%, 5.4%, respectively) (all P<0.001). (3) The main clinical manifestations were fever (63.5%), respiratory symptoms (53.7%) and lympha-denopathy (53.7%), 47.5% of the children with hyper blood IgE had an increased white blood cell count, and 12.1% of them had an increased eosinophil count.(4) The most common specific allergens were dust mite combination (32.0%), milk (17.0%), and egg white (16.0%). There was no difference in disease distribution among the 3 groups of hyper blood IgE children with positive specific IgE ( P=0.164). Conclusions:Hyper blood IgE in children are most commonly caused by allergic and infectious diseases.The etiological distributions of hyper blood IgE in children at varying severities differ a lot.The higher the total IgE level, the higher the incidence of immunodeficiency disease, rheumatic disease, tumor and hematological disease.