Anterior cruciate ligament is the most important ligament to maintain the anterior and rotation stability of knee joint. Rupture of anterior cruciate ligament is one of the most common injuries of knee joint, and thus leads to knee instability and traumatic osteoarthritis. Anterior cruciate ligament reconstruction is usually performed to restore the anterior stability of knee joint, and is considered to reduce the secondary injury of medial meniscus, lateral meniscus and cartilage. Thus anterior cruciate ligament reconstruction can improve the function of knee joint. Traditional single bundle technique to reconstruct anterior cruciate ligament has been performed for many years. This technique can restore the anterior stability of knee joint and has excellent clini?cal results. Nearly 61%patients showed obvious radiographic osteoarthritis 20 years after anterior cruciate ligament reconstruction using bone-patella-bone graft. But, there is no agreement regarding to reduce the development of osteoarthritis after reconstruc?tion of anterior cruciate ligament. However, it has been reported that osteoarthritis would develop after reconstruction of anterior cruciate ligament in long term follow up study. Recently, with the further understanding of anatomy of biomechanics of anterior cru?ciate ligament, new techniques for anterior cruciate ligament reconstruction are developed, such as double bundle reconstruction, anatomic reconstruction and individual reconstruction. It remains controversial that whether these new technique can prevent the development of osteoarthritis after rupture of anterior cruciate ligament. Currently, no reconstruction technique for anterior cruci?ate ligament is perfect, and every technique has advantages and disadvantages. In terms of reducing the prevalence of osteoarthri?tis after reconstruction of anterior cruciate ligament, which technique is the best still remains unclear. New treatment and evalua?tion methods should be developed. In the future, not only the restoration of stability of knee joint should be considered, but also the articular cartilage contact kinematics including tibiofemoral joint and patellofemoral joint after anterior cruciate ligament recon?struction. Reduction of the development of osteoarthritis is an important topic after reconstruction of anterior cruciate ligament.

Objective To study the expression of hTERT protein in precancerous lesions of human stomach, and to analyze the relationship between telomerase activity and cell apoptosis, in order to elucidate the effects of telomerase activity and cell apoptosis in the pathogenesis of gastric carcinoma. Methods Sp method was employed to detect the hTERT expression and the apoptosis was determined in situ by TUNEL staining in 64 patients with chronic gastritis and gastric carcinoma. Among the 64 patients, 16 suffered from chronic gastritis with gastratrophia, 15 with intestinal metaplasia, 14 with allotypic proliferation, and the 19 remainders suffered from gastric carcinoma. Results (1) The respective positive frequency of hTERT protein expression in chronic gastritis, intestinal metaplasia, gastric allotypic proliferation and human gastric carcinoma were 25.0%, 46.7%, 64.3% and 78.9%. The positive frequency of hTERT protein expression in human gastric carcinoma was significantly higher than that in gastric allotypic proliferation, intestinal metaplasia and chronic atrophic gastritis (P

Objective To study the expression of hTERT in human gastric carcinoma and its precancerous lesions and to explore the relationship between hTERT expression and Helicobacter pylori (Hp) infection. Methods hTERT expression was detected by immunohistochemical method. Hp was detected by rapid urease test and Warthin-Starry silver staining. Results The expression level of hTERT in human gastric carcinoma (78.95%) was significantly higher than that in gastric dysplasia (64.29%), in intestinal metaplasia (46.67%), and in atrophic gastritis (25.00%, P

Objective To assess the morphological characteristics of bone marrow in patients of severe fever with thrombocytopenia syndrome ( SFTS) and its value in diagnosis.Methods The bone marrow morphology was retrospectively reviewed in 28 laboratory confirmed patients with SFTS from Zhoushan Hospital during January 2012 and December 2015.The correlation between bone marrow -derived macrophage and peripheral blood cells was analyzed with t test.Results All patients presented leukocytopenia and thrombocytopenia.Poor bone marrow hematopoietic function was observed in 23 patients (82%) showing granulocyte, erythrocyte and megakaryocyte hypoplasia , but no pathological hematopoietic disorder was observed.Eighteen patients (64%) had various degrees of increased amount of macrophage in the bone marrow; peripheral white blood cell count and platelets in patients with macrophage ≥0.5% were lower than those with macrophage <0.5%, and the difference was of statistical significance (t =3.836 and 4.499, P<0.01).Conclusion SFTS patients have characteristic bone marrow morphology , and bone marrow examination is beneficial for differentiation of SFTS from blood lymphatic system diseases and other virus infection.

Objective To analyze the relationship between cytotoxic T cells Perforin level, IFN-γand IL-10 in patients with chronic hepatitis B. Methods After a short-term cultivation of peripheral blood collected from 50 patients with chronic hepatitis B, a flow cytometry was employed to detect the levels of Per-forin, IFN-γ and IL-10 in CD8~+ cells to compare with those of the health donors the HBV DNA copies in their blood were also measured by RT-PCR to analyze the relationship with Perforin, IFN-γ, and IL-10 in CD8~+ cells. Results In patients with chronic hepatitis B, the levels of Perforin, IFN-γ and IL-10 in CD8~+ ceils were (5.30 ± 2.62)%, (4.05 ± 2.25) % and (0.77 ± 0.50) %, respectively, which were statistical-ly lower than health donors(the t values were 4.50, 4.56 and 4.20 respectively; P < 0.01) ; Of 26 cases of chronic hepatitis B patients with HBeAg-positive Pefforin and IFN-γ in CD8~+ T cells were (4.54 ± 1.93) % and (3.32 ± 1.59)%, respectively, significantly lower than those of the 24 chronic hepatitis B patients with HBeAg-negative (the t values were 2.22 and 2.54, respectively; P <0.05) ; And the levels of Perforin, IFN-γ had a negative relation with the HBV DNA copies (the coefficient correlations-0. 539 and-0. 340; P < 0.01 and P < 0.05). Conclusion In patients with chronic hepatitis B, the reduced levels of Pefforin, IFN-γ and IL-10 may be related with the long-term existence of HBV, and the protracted course of disease.

Objective To investigate the significance of measurement of the expression CD28 on tumor infiltrating lymphocytes in patients with stomach cancer.Methods Subjects for this investigation include the tumor tissues and its nearby normal tissues from 48 patients with stomach cancer.The levels of the expression of CD28, CD8 on tumor infiltrating lymphocytes were analyzed by flow cytometry.Results First, the levels of the expression of CD8+CD28+ and CD8-CD28+ on tumor infiltrating lymphocytes from the tumor tissues were remarkably lower than from its nearby normal tissues (P

BACKGROUND: Meseuchymal stem cells (MSCs) have extremely strong self-duplication ability and multidirectional ifferentiation potential. When bone marrow stromal cells (BMSC) are isolated and cultured in vitro, implanted in vivo, the distribution and colonization are still unclear, which is concerned with whether BMSC can be usedas target cells in clinic.OBJECTIVE: To explore the capacity of colonizing to the liver after allografting of green fluorescent protein (GFP) labeled MSCs of rats by different approaches.DESIGN: Factorial design.SETTING: Department of General Surgery, Second People's Hospital of Guangdong Province, Postdoctoral Workstation of Sun Yat-Sen University;Department of General Surgery, Zhujiang Hospital, Southern Medical University; Department of Organ Transplantation, Third Hospital Affiliated to Sun Yat-Sen University.MATERIALS: The experiment was performed at the Staff Room of Pharmacology, Basic Department, First Military Medical University of Chinese PLA from January 2003 to December 2004. A total of 36 clean adult SD rats were selected and randomly assigned into 5 groups: CCL4 plus portal vein transplantation group (n=6), portal vein transplantation control group (n=6), CCL4 plus caudal vein transplantation group (n=6), caudal vein transplantation control group (n=6) and mixed group (n=12).METHODS: ① MSCs were obtained from rat marrow and labeled with GFP. After amplifying in vitro, MSCs suspension was implanted with thin needle, with the volume of 0.5 mL/100 g. ②CCL4 plus portal vein transplantation group: In 3 days before MSCs transplantation, the rats were administrated with 20 g/L CCL4 2.5 mL/kg by gastric perfusion every day. The dose was double at the first time. Labeled MSCs were implanted from portal vein. Portal vein transplantation control group: Before transplantation the MSCs were bred commonly, and the labeled MSCs were implanted from portal vein. CCL4 plus caudal vein transplantation group: In 3 days before MSCs transplantation, the rats were administrated with 20 g/L CCL42.5 mL/kg by gastric perfusion every day. The dose was double at the first time. Labeled MSCs were implanted from caudal vein. Caudal vein transplantation control group: Before transplantation the MSCs were bred commonly, and the labeled MSCs were implanted from caudal vein. Mixed group: On the basis of the former 4 groups, 2 rats were implanted with non-labeled MSCs; Another 2 rats fed with CCL4 for 3 days and normal feed were established, without MSCs transplantation. ③At days 3 and 7 after transplantation expression of transplanted MSCs in liver of rats of each group were examined with fluorescent quantitative PCR.MAIN OUTCOME MEASURES: ①Results of MSCs isolation, purification, in vitro amplification and phenotype identification, ②result of GFP-labeled MSCs, ③observation of growth of rats following allografting of MSCs, and ④result of quantitative identification of GFP positive DNA amount in hepatic tissues of each group.RESULTS: Totally 36 experimental SD rats were involved in the result analysis. ①Percoll gradient separating medium was applied to isolate bone marrow of rats. The obtained cells were transferred and amplified,and then mostly showed coincident shuttle shape. Cells did not express CD34 and CD45, but CD29, CD44 and CD90 of MSCs, which were noncommitted stem cells in non-differentiating status that were different from hemopoietic stem cells in bone marrow. ②The green fluorescent cells appeared 24 hours after MSCs transfection. From hour 48 to 72 the number of positive cells significantly increased, with strong intensity.The transfection efficiency was 20%-30% under high-power field, and most of the cells were with green fluorescence. But green fluorescent cells did not appear in the MSCs cells as control. ③After allografting of labeled or non-labeled MSCs of rats with different approaches, at day 1 the rats were listless with bad food appetite, less mobilization; At day 2mostly of them had normal diet and mood, but there was no significant difference in rats of each group. ④The rats in each group with the exception of mixed group had green fluorescent protein positive cells in liver at days 3 and 7. The number of green fluorescent protein positive DNA was higher in liver tissues in the CCL4 plus portal vein transplantation group and CCL4 plus caudal vein transplantation group than in the portal vein transplantation control group and caudal vein transplantation control group (P＜0.05).CONCLUSION: Duration and amount of stem cells colonizing in liver may be associated with liver injury, while not related to the implantation approach. In normal animals with uninjured liver the stem cells can colonize in liver, and the amount is associated with transplantation approach and post-transplantation duration.

Objective:To evaluate the effects of intranasal administration of glial cell line-derived neurotrophic factor (GDNF) on postoperative cognitive dysfunction in aged rats.Methods:Forty healthy Sprague-Dawley rats of both sexes, aged 21-23 months, weighing 480-600 g, were divided into 4 groups ( n=10 each) using a random number table method: sham operation group (group S), operation group (group O), intranasal administration of low-dose GDNF group (group G1) and intranasal administration of high-dose GDNF group (group G2). Rats underwent exploratory laparotomy under anesthesia with chloral hydrate in O, G1 and G2 groups, while the rats in group S only received sham operation.The rats in group G1 and group G2 were intranasally treated with GDNF 25 and 50 μg (in 25 μl of PBS), respectively, and PBS 25 μl was nasally administered in group S and group O every day for 3 consecutive days after operation or sham operation.Morris water maze test was performed on days 3-7 after surgery, and then the rats were sacrificed, and hippocampal tissues were removed for determination of the expression of GDNF, high mobility group box 1 (HMGB1), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), activated caspase-3 and Bax (by Western blot). Results:Compared with group S, the escape latency was significantly prolonged on days 5-7 after operation, the number of crossing the platform was reduced, time spent in the target quadrant was shortened, expression of GDNF was down-regulated, and expression of IL-1β, TNF-α, HMGB1, activated caspase-3 and Bax in hippocampi was up-regulated in group O, and the number of crossing the platform was reduced, time spent in the target quadrant was shortened, and expression of IL-1β and TNF-α was up-regulated in G1 and G2 groups ( P<0.05). Compared with group O, the escape latency was significantly shortened on days 5-7 after operation, the number of crossing the platform was increased, time spent in the target quadrant was prolonged, expression of GDNF was up-regulated, expression of TNF-α, HMGB1, activated caspase-3 and Bax in hippocampi was down-regulated in G1 and G2 groups, and IL-1β in hippocampi was down-regulated in group G1 ( P<0.05). Compared with group G1, the expression of TNF-α in hippocampi was down-regulated ( P<0.05), and no significant change was found in the other parameters mentioned above in group G2 ( P>0.05). Conclusions:Intranasal administration of GDNF can improve postoperative cognitive dysfunction, and the mechanism may be related to inhibiting neuroinflammatory responses and neuroapoptosis in aged rats.

Objective To evaluate the relationship between α7 nicotinic acetylcholine receptor (α7nAChR) signaling pathway and regulatory T cells (Tregs) during vagus nerve stimulation-induced reduction of endotoxin-caused acute lung injury (ALI) in mice.Methods Clean-grade healthy male C57BL/6 mice,aged 6-8 weeks,weighing 22-25 g,were divided into 5 groups (n=10 each) using a random number table method:control group (group C),group ALI,vagus nerve stimulation group (group VNS),α-BGT group (group α-BGT) and vagus nerve stimulation plus α-BGT group (group VNS + α-BGT).After successful establishment of the model,the vagus nerve was stimulated for 30 s with a stimulus intensity of 0.5 mA,frequency of 20 Hz,an interval of 5 min,60 min in total.Sterile normal saline 100 μl was injected into the trachea,and the vagus nerve was only exposed but not stimulated in group C.In group ALI,the ALI model was established,and the vagus nerve was isolated but not stimulated.In group α-BGT,α-BGT 1 μg/kg was intraperitoneally injected,the ALI model was established 1 h later,and the vagus nerve was isolated but not stimulated.In group VNS+α-BGT,α-BGT 1 μg/kg was intraperitoneally injected,the ALI model was established 1 h later,and the vagus nerve was stimulated at 1 h after the end of establishment.Animals were sacrificed at 72 h after establishing the model,and lungs were removed for determination of lung water content,percentage of Tregs (using flow cytometry),myeloperoxidase (MPO) activity (by colorimetric assay),expression of α7nAChR (by Western blot) and contents of interleukin-10 (IL-10),transforming growth factor-β (TGF-β) and IL-1β (by enzyme-linked immunosorbent assay).Results Compared with group C,the lung water content,MPO activity and IL-1β content were significantly increased in the other four groups,the expression of α7nAChR was significantly down-regulated in ALI,α-BGT and VNS+α-BGT groups,the percentage of Tregs was significantly increased in group VNS,the IL-10 content was significantly decreased in ALI and α-BGT groups and increased in VNS and VNS+α-BGT groups,and TGF-β contents were significantly decreased in ALI,α-BGT and VNS+α-BGT groups and increased in group VNS (P＜0.05).Compared with group ALI,the lung water content,MPO activity and IL-1β content were significantly decreased,the expression of α7nAChR was up-regulated,and the percentage of Tregs and contents of IL-10 and TGF-β were increased in group VNS,and the TGF-β content in group α-BGT and contents of IL-10 and TGF-β in group VNS+α-BGT were significantly increased (P＜0.05).Compared with group VNS,the lung water content,MPO activity and IL-1β content were significantly increased,the expression of α7nAChR was down-regulated,and the percentage of Tregs and contents of IL-10 and TGF-β were decreased in α-BGT and VNS+α-BGT groups (P＜0.05).The contents of IL-1O and TGF-β were significantly higher in group VNS+α-BGT than in group α-BGT (P＜0.05).Conclusion Vagus nerve stimulation can activate α7nAChR signaling pathway and raise the percentage of Tregs,thus reducing ALI in mice.

Objective:To evaluate the effect of glucagon-like peptide 1 and glucose-dependent insulinotropic polypeptide receptor agonist DA-JC4 on postoperative neuroinflammatory responses in aged rats.Methods:Forty-five Sprague-Dawley rats, aged 21-23 months, weighing 530-630 g, provided by the Animal Experiment Center of Medical School of Zhengzhou University, were assigned into 3 groups ( n=15 each) using a random number table method: sham operation group (group S), operation group (group O) and DA-JC4 group (group G). Rats underwent exploratory laparotomy under anesthesia with chloral hydrate in O and G groups.In group G, DA-JC4 10 nmol/kg (dissolved in 1 ml of sterile normal saline) was intraperitoneally injected immediately after the end of operation and at 24 and 48 h after operation.Western blot was used to determine the expression of hippocampal Bax, Bcl-2, activated caspase-3, Beclin-1, microtubule-associated protein 1 light chain 3Ⅱ (LC3Ⅱ), high-mobility group box 1 protein (HMGB1), interleukin-1beta (IL-1β) and tumor necrosis factor-alpha (TNF-α) on day 3 after surgery.The Morris water maze test was performed on days 14-18 after operation to assess the cognitive function. Results:Compared with group S, the escape latency was significantly prolonged on days 15-18 after operation in group O and on day 18 after operation in group G, and the number of crossing the original platform was reduced, the time spent in the target quadrant was shortened, the expression of activated caspase-3, Bax, LC3Ⅱ, HMGB1, IL-1β and TNF-α in hippocampi was up-regulated, and the expression of Bcl-2, LC3Ⅱ and Beclin-1 was down-regulated in O and G groups ( P<0.05). Compared with group O, the escape latency was significantly shortened, and the number of crossing the original platform was increased, the time spent in the target quadrant was prolonged, the expression of activated caspase-3, Bax, LC3Ⅱ, HMGB1, IL-1β and TNF-α in hippocampi was down-regulated, and the expression of Bcl-2, LC3Ⅱ and Beclin-1 was up-regulated in group G ( P<0.05). Conclusion:The mechanism by which DA-JC4 reduces postoperative cognitive dysfunction may be related to inhibiting neuroinflammatory responses in aged rats.