Objective To investigate the expression level of endogenetic nitric oxide(NO) in the reproduction process of human anterior cruciate ligament cells. Methods Anterior cruciate ligament cells were isolated and subcultured from the human anterior cruciate ligament. LPS was used to induce the anterior cruciate ligament cell to express the inducible nitric oxide synthase(iNOS ), and N-monomethyl -L-Arginine (L-NMMA) was used as the interdiction of nitric oxide. They were alone or together added in the culture medium of anterior cruciate ligament cells in different groups. The level of NO was indirectly measured in the medium of HACL. Results LPS promoted significantly anterior cruciate ligament cells to produce endogenetic nitric oxide, compared with the control group(P

BACKGROUND: Intra-articular injection of hormone has been applied in the treatment of arthritis because it can alleviate arthralgia rapidly, which is accompanied commonly by progressive cartilage impairments. It is not clear if supplement of growth factor like insulin effect can play a protective role in articular chondrocytes.OBJECTIVE: To observe the effects of insulin or hydrocortisone alone and the combination on the proliferation of chondrocytes.DESIGN: Grouping comparative study, the effect of one medicine was analyzed by using one-factor analysis of variance, while the combined effect was analyzed with multi-factor analysis of variance.SETTING: Guangdong Institute of Trauma Sugery.MATERIALS: Articular cartilage from the knees of New Zealand white rabbits of 4 - 6 weeks old.METHODS: This study was carried out at Guangzhou Traumatic Research Institute from Feberary 2000 to May 2001. Chondrocytes were isolated from the knee joints of New Zealand white rabbits, digested with hyaluronidase,pancreatin and type Ⅱ collagenase and exposed to insulin, hydrocortisone or the combination of insulin and hydrocortisone of different dosage. They were divided into four groups:Control group ( without adding insulin and hydrocortisone), insulin group (0. 035,0. 35,3.5,35 mg/L subgroups), hydrocortisone group(1,5,10,50,100 mg/L subgroups) and insulin(0. 35 mg/L) combined with hydrocortisone(50 mg/L) group. Their influence on chondrocytes proliferation was observed with methyl thiazolyl tetrazolium(MTT) method.sulin.at the concentration of 0. 035 mg/L( P ＜ 0.01 ), reaching the maximum at could inhibit the proliferation of chondrocytes ( P ＜ 0.05 ), which became significant with increasing concentration and no viable chondrocytes could be exposed to 0 . 35 mg/L insulin combined with 50 mg/L hydrocortisone, the promoting effect of insulin was inhibited due to negative cooperation.CONCLUSION: Insulin at low concentration could enhance the proliferation of chondrocytes, but hydrocortisone displayed inhibiting effect on the growth of chondrocytes. The function of insulin was antagonized when combined with hydrocortisone.

BACKGROUND: Type Ⅱ collagen has been used as the carrier for chondrocyte transplantation in animal models, but whether type Ⅱ collagen may cause arthritis or mediate cytotoxicity remains unknown.OBJECTIVE: To detect the cellular immune functions of the New Zealand rabbits immunized by porcine type Ⅱ collagen.DESIGN: An exploratory comparative study based on the observations.SETTING: An institute of trauma surgery of a municipal hospital.MATERIALS: The study was conducted in the Institute of Trauma Surgery,Guangzhou Red Cross Hospital from August 1999 to February 2000. Six New Zealand rabbits, whose body mass ranged from 2.0 kg to 3.0 kg, were chosen of either gender.METHODS: The rabbits were immunized by porcine type Ⅱ collagen for 60days, during which the plasma was regularly taken for detection of type Ⅱ collagen antibody. On the 60th day, the peripheral blood as well as the spleens and lymph nodes were taken to separate the lymphocytes, which were subjected to secondary stimulation with type Ⅱ collagen in vitro to observe the reactive cell proliferation. The lymphocytes were randomly divided into two groups, and the first group was treated with phytohemagglutinin(PHA) of different concentrations to serve as the positive control, in which non-specific immunity was examined; The second group was treated with type Ⅱ collagen of different concentrations for examining specific immunity.peripheral blood lymphocytes of normal and immunized rabbits.RESULTS: On the 21st day, the titer of the antibody presented the first peak, and 40 days after the re-injection of the antigen the second peak appeared, which maintained for 20 days and then gradually descended. The lymphocytes of the normal rabbits proliferated in response to PHA stimulation but not to the first stimulation with the type Ⅱ collagen. The lymphocytes of the immunized rabbits exhibited significant proliferation upon stimulations with both PHA and type Ⅱ collagen. At the concentration of 25 mg/L, type Ⅱ collagen stimulation was sufficient to induce lymphocyte proliferation, the peak of which occurred when the collagen concentration reached 50 mg/L.CONCLUSION: Xenogenic type Ⅱ collagen at an adequate concentration may induce the increase of the type Ⅱ collagen antibody in immunized rabbits and proliferation of lymphocytes of the spleens and peripheral blood to cause cellular immune reaction and even immunological arthritis in relation to the transplantation.

OBJECTIVETo culture fibroblast cells from the knee ligaments and to study the biological characteristics of these cells.

METHODSCells of the anterior cruciate ligament (ACL) and the medial collateral ligament (MCL) from New Zealand white rabbit were cultured in vitro. Cellular growth and expression of the collagen were analyzed. Moreover, an in vitro wound closure model was established and the healing of the ACL and the MCL cells was compared.

RESULTSMaximal growth for all these cells were obtained with Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum, but RPMI 1640 and Ham's F12 media were not suitable to maintain these cells. Morphology of both ACL and MCL cells from New Zealand white rabbit was alike in vitro, but the MCL cells grew faster than the ACL cells. Both cell types produced similar amount of collagen in culture, but the ratio of collage type I to type III produced by ACL cells was higher than that produced by MCL cells. Wound closure assay showed that at 36 hours after injury, cell-free zones created in the ACL cultures were occupied partially by the ACL cells; in contrast, the wounded zone in the MCL cultures was almost completely covered by the cells.

CONCLUSIONSAlthough the ACL cells and the MCL cells from New Zealand white rabbit show similar appearance in morphology in culture, the cellular growth and the biochemical synthesis of collagen as well as the healing in vitro were significantly different. These differences in intrinsic properties of the two types of cells in vitro might contribute to the differential healing potentials of these ligaments in vivo.

Animals ; Anterior Cruciate Ligament ; cytology ; Cell Division ; physiology ; Cells, Cultured ; Collagen ; metabolism ; Collateral Ligaments ; cytology ; Culture Media ; Female ; Fibroblasts ; physiology ; Male ; Rabbits ; Sensitivity and Specificity

To detect the cellular immunity state of New Zealand white rabbit immunized by pig type II collagen. The New Zealand white rabbit was immunized by type II collagen for sixty days. The plasma was collected at a regular interval and the anti-type II collagen antibodies were examined. At the sixtieth day, the peripheral circular lymphocytes and the lymphocytes separated from spleen cells of rabbit and lymph nodes were collected and were stimulated by type II collagen in vivo again. The regulation of reactive cellular proliferation caused by the stimulation was detected. The experiment samples were divided into two groups. The first group was the positive control group by adding different concentrations of PHA and the non-specific immunity was assayed. The different concentrations of type II collagen were added to the second group and the specific immunity was assayed. The lymphocytes of normal rabbits showed proliferation by PHA stimulation but no proliferation by the first stimulation of type II collagen. Obvious proliferation due to the stimulation of both PHA and type II collagen in the immunized rabbit were observed. It shows that certain concentration of heterogeneous collagen may cause an increase of anti-type II collagen antibody in immunized rabbit and may cause a proliferation of lymphocytes in rabbit spleen and peripheral blood. The heterogeneous type II collagen causes cellular immunity in vivo.
Animals ; Cell Division ; drug effects ; Collagen Type II ; administration & dosage ; immunology ; Cytotoxicity, Immunologic ; Female ; Histocompatibility Antigens ; Lymphocytes ; cytology ; immunology ; Male ; Rabbits ; Spleen ; cytology ; Swine ; Transplantation, Heterologous

Objective:To explore the application effect of the "4M1E" comprehensive management mode (man, machine, material, method, environment) in ultrasonic examination, and to provide reference for the health management center of hospitals to formulate reform measures to improve the quality and efficiency of ultrasonic examination.Methods:By adoping a non-randomized controlled research, a total of 400 clients who underwent physical examination in the Health Management Center of Zhujiang Hospital, Southern Medical University were selected as subjects by convenience sampling method. Among them, 200 clients who underwent physical examination from August to October 2021 were selected as the control group to implement routine physical examination mode, another 200 clients who underwent physical examination from August to October 2022 were selected as the experimental group to implement the "4M1E" comprehensive management mode. The ultrasonic examination time, passing rate, bladder filling, pre-examination knowledge score and satisfaction of the two groups were compared.Results:The ultrasonic examination time of experimental group was (9.54 ± 3.25) min, lower than (12.33 ± 5.07) min of control group, and the difference was statistically significant ( t=6.53, P<0.05). The passing rates of ultrasonic examination at 1 time, 2 times and 3 times in the experimental group were 86.0% (172/200), 12.0% (24/200) and 2.0% (4/200), respectively, which were better than 73.5% (147/200), 20.5% (41/200) and 6.0% (12/200) in the control group with statistical significance ( χ2=10.05, P<0.05); the proportions of overfilling, proper filling and poor filling in the experimental group were 1.0% (2/200), 86.0% (172/200) and 13.0% (26/200), respectively, which were better than 2.0% (4/200), 70.0% (140/200) and 28.0% (56/200) in the control group, the differences of two groups were statistically significant ( χ2=14.93, P<0.05). Pre-examination knowledge score of experimental group (8.16 ± 1.43) was higher than that of control group (6.19 ± 1.94) points, and the difference was statistically significant ( t=-11.48, P<0.05). The overall satisfaction score of the experimental group (20.55 ± 2.46) was higher than that of the control group (18.45 ± 2.86). The differences was statistically significant ( t=-7.87, P<0.05). Conclusions:The "4M1E" comprehensive management mode can make physical examination clients fully prepare for the examination, reduce the time of ultrasonic examination, complete the examination efficiently, and comprehensively improve the sense of experience and satisfaction.

Background: and Purpose: Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is a severe central nervous system disorder mediated by NMDAR antibodies that damages neurons. We investigated the correlation between cytoskeletal autoantibodies and the clinical severity in patients with anti-NMDAR encephalitis. Methods: Non-NMDAR autoantibodies were identified by screening matched cerebrospinal fluid (CSF) and the serum samples of 45 consecutive patients with anti-NMDAR encephalitis and 60 healthy individuals against N-methyl-D-aspartate receptor 1-transfected and nontransfected human embryonic kidney 293T cells. Immunocytochemistry was performed to assess antibody binding in rat brain sections and primary cortical neurons. Cell-based assays and Western blotting were applied to identify autoantibodies targeting medium neurofilaments (NFMs). We compared clinical characteristics between patients with NMDAR encephalitis who were positive and negative for anti-NFM-autoantibodies. Results: Anti-NFM autoantibodies were detected in both the serum and CSF in one patient (2%) and in the serum only in six patients (13%). No antibodies were detected in the serum of healthy controls (7/45 vs. 0/60, p=0.0016). Four of the seven patients with anti-NFM autoantibodies in serum were children (57%), and three (43%) had abnormalities in brain magnetic resonance imaging. These patients responded well to immunotherapy, and either no significant or only mild disability was observed at the last follow-up. Anti-NMDAR encephalitis did not differ with the presence of anti-NFM autoantibodies. Conclusions Anti-NFM autoantibodies may be present in patients with anti-NMDAR encephalitis, indicating underlying neuronal damage. A large cohort study is warranted to investigate the clinical differences between patients with NMDAR encephalitis according to their antiNFM antibody status.

Objective:To understand the epidemiological and clinical characteristics of melioidosis in Haikou City, to rise the people's awareness of melioidosis and to provide basis for prevention and control of the disease.Methods:The clinical data of 254 patients with melioidosis treated in 4 Class A tertiary hospitals in Haikou City from January 2000 to September 2020 were collected, and the epidemiological characteristics, clinical manifestations, infection site, prognosis and drug sensitivity were retrospectively analyzed.Results:Among 254 patients with melioidosis, 226 males (88.98%) and 28 females (11.02%), and the gender ratio was 8.07 ∶ 1.00. Farmers were the main occupation, accounting for 37.80% (96/254). The median age was 53 years old, mainly in 41 - 80 years old, accounting for 83.46% (212/254). Han nationality was the most, accounting for 89.76% (228/254). The onset season was mainly in summer and autumn, and the peak was from August to October (117 cases). Patients were mainly distributed in coastal areas, among which Haikou City (49 cases) was the most, followed by Dongfang City (46 cases), Danzhou City (23 cases) and Wenchang City (21 cases). Totally 196 cases (77.17%) had basic diseases, diabetes was the most common (162 cases). The main symptoms of admission were fever (211 cases), followed by cough (108 cases) and expectoration (88 cases). The infection sites were mainly blood (104 cases, 40.94%), lung (60 cases, 23.62%), liver and spleen (32 cases, 12.60%). Totally 195 patients were treated with sensitive antibiotics, at discharge, 37 cases (18.97%) were cured, 129 cases (66.15%) improved, 18 cases (9.23%) did not heal, 7 cases (3.59%) died and 4 cases (2.05%) were discharged voluntarily. Results of drug sensitivity tests from 2010 to 2020 showed that the sensitivity rates of Burkholderia pseudomallei to imipenem (142 cases), meropenem (16 cases) and ceftazidime (141 cases) were all 100.00%, and the sensitivity rates of doxycycline (25 cases) and compound sulfamethoxazole (142 cases) were 92.00% (23/25) and 99.30% (141/142), respectively. Conclusions:Males, farmers, middle-aged and elderly people and people with diabetes and other basic diseases are the high incidence population of melioidosis in Haikou City. The incidence peak is in summer and autumn. The common clinical manifestations are fever, pulmonary infection, abscess of liver and spleen, etc. In the treatment, Burkholderia pseudomallei is more sensitive to imipenem, meropenem and ceftazidime.