Objective: To express glycosyl-phosphatidylinositol (GPI) modified Met- RANTES fusion protein on Chinese hamster ovary (CHO) cells and to develop a novel immunosuppressant GPI anchored form of Met-RANTES. Methods: The eukaryotic expression vector PEF/GPI-Met-RANTES were constructed and transfected into CHO cells by electroporation. The transfectants were selected with methotrexate (MTX). Expression of the recombinant protein was assessed by flow cytometric analysis, cell immunofluorescence staining and immunogold electron microscopy. Results: The chimeric molecules of GPI anchored form of Met-RANTES including the whole reading frame were constructed, and the sequence was identical to the designed sequence. GPI anchored form of Met-RANTES was stably expressed on CHO- DHFR- cells. Conclusion: A large amount of GPI modified Met-RANTES fusion protein was expressed on CHO cells. GPI anchored form of Met-RANTES may be used as novel immunosuppressant for suppressing reaction in graft rejection.

As the key phase for medical students to be doctors,clinical internship plays a vital role in the whole medical education process.Interns in the department of gastrointestinal surgery face series of difficulties,such as wide variety of diseases and emergence of new technology and new concepts.In addition,because of the uneven levels of teachers and low enthusiasm of interns,the result of clinical practice teaching is not satisfactory.Based on the above-mentioned problems,measures have been introduced during the process of clinical practice teaching,including selection of typical diseases and surgical procedures,construction of clinical practice teaching team,adoption of pluralistic teaching methods,reinforcement of participation of interns during the clinical practice.Finally,the quality of teaching in our department has been irmproved significandy.

AIM To establish an HPLC method for the simuhaneous content determination of five flavonoids in Duchesnea indica (Andr.) Focke at five picking time (April,May,June,July and August).METHODS The analysis of D.indica methanol extract was performed on a 25 ℃ thermostatic Agilent ZORBAX SB-C18 column (250 mm×4.6 mm,5 μm),the mobile phase comprising of acetointrile-0.1％ methanoic acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 350 nm.RESULTS Rutin,hyperoside,isoquercitrin,celereoin and kaempferol showed good linear relationships within their own ranges (R2 ≥0.998 6),whose average recoveries were 97.1％-101.5％ with the RSDs of 1.37％-2.37％.The contents of five constituents in samples at different picking time exhibited obvious differences,among which lutin and hyperoside contents were the highest in June,isoquercitrin content was the highest in July,and celereoin and kaempferol contents were the highest in August.CONCLUSION The suitable picking time of D.indica is June and July.

Objective To discuss the value of CT prognosis on the Masaoka staging system of thymic epithelial tumors(TET) before surgical resection.Methods The CT images of 102 patients with TET proved by surgery and pathology were reviewed retrospectively.The TET were reclassified according to Masaoka stage system.The size,homogeneity,sharp,contour,infiltration of surrounding tissue,and metastasis on CT were analyzed with Logistic analysis.The diagnostic value was also evaluated with a ROC curve.Results Masaoka pathologic stages were stage Ⅰ for 36 (35.3 ％),stage Ⅱ for 27 (26.5 ％),stage Ⅲ for 30 (29.4 ％),and stage Ⅳ for 9 (8.8 ％).A multivariable Logistic regression model showed that TET with larger size of tumor (20/35,P =0.0371,OR =4.539),irregular or lobulated tumor contour (26/42,P =0.0230,OR =4.870),heterogeneous (21/33,P =0.0154,OR =6.020),infiltration of surrounding fat (25/32,P =0.0019,OR =14.005),and pleural seeding (11/11,P =0.0032,OR =36.153)were more likely to have stage Ⅲ or Ⅳ disease.The area under ROC curve was 0.940.Conclusions The tumor CT imaging features can differentiate between stage Ⅰ,Ⅱ and stage Ⅲ,Ⅳ disease.This helps identified patients more likely to benefit from neoadjuvant therapy.

Abstract: Objective　To explore the effect of Yiaikang on the expression of sterile alpha motif and histidine-aspartic acid domain-containing protein 1(SAMHD1), a host restriction factor, and interferon levels in peripheral blood of HIV-infected patients with lung-spleen Qi deficiency syndrome. This aims to provide insights into the mechanism of YiaiKang in the treatment of AIDS. Methods According to the diagnostic criteria of Western medicine for AIDS and traditional Chinese medicine for lung-spleen Qi deficiency syndrome, 22 cases of HIV patients with lung and spleen Qi deficiency syndrome were enrolled in the study from the First Affiliated Hospital of Henan University of TCM between September 2020 and May 2021, and 20 healthy people in the area were taken as control. The patients with lung and spleen Qi deficiency syndrome were treated with Yiaikang for 6 months. T lymphocyte subpopulation changes and expression level of SAMHD1 protein in T cells were detected by flow cytometry. SAMHD1 mRNA expression in peripheral blood mononuclear cells (PBMCs) was detected by fluorescence quantitative PCR. The levels of IFN-α, IFN-β, IFN-γ, and IL-2 in plasma were detected by enzyme-linked immunosorbent assay (ELISA). The changes of SAMHD1 and interferon in peripheral blood of HIV patients with lung and spleen Qi deficiency syndrome were analyzed to explore the intervention effect of Yiaikang. Results Compared with healthy people, HIV patients with lung and spleen Qi deficiency syndrome exhibited a reduction in the CD4 + T lymphocyte count and CD4/CD8 ratio (Z=-3.526, P<0.001; Z=-0.881, P<0.001). There was an increase in the expression of SAMHD1 protein in T lymphocytes (SAMHD1 MFI/CD4: Z=-3.425, P=0.001; SAMHD1 MFI/CD8: Z=-2.153, P=0.031) and an increase in the expression of SAMHD1 gene in PBMCs (Z=-3.828, P<0.001). Yiaikang treatment significantly increased the CD4 + T cell count and CD4/CD8 ratio (Z=-2.711, P=0.007; Z=-2.062, P=0.039), decreased the expression of SAMHD1 mRNA (Z=-2.581, P=0.010), and decreased the levels of IFN-α and IFN-γ (Z=-1.985, P=0.047; Z=-2.744, P=0.006). Conclusions Patients with lung-spleen Qi deficiency syndrome have reduced numbers of CD4 + T cells and CD4/CD8 ration, with increased expression of SAMHD1 and interferon factors. Yiaikang treatment can increase the number of CD4 + T cells and CD4/CD8 ratio and reduce the expression of SAMHD1 and interferon factor, thereby improving the immune imbalance caused by HIV infection.

Objective To evaluate the effect of transverse position and numbers on the stability of the spinal pedicle screw fixation during the pedicle cortex perforation. Methods The vertebral compression fracture was performed in the L1 vertebral body using the Chiba's method from 60 fresh thoracic and lumbar vertebrae samples of sheep(T13-L3),which were randomly divided into 6 groups(A,B,C,D,E,and F)using a lottery method. Bilateral pedicles of vertebral arch of T14 and L2 were inserted pedicle screws,connecting titanium rods to fix T14-L2 segments. Then the samples of groups B,C,D,E,and F were removed a quarter of right side of lateral T14 thoracic pedicle cortical,which were considered the pedicle cortex perforation model. Finally,each group was fixed on different numbers of crosslinks:group A(0 crosslink,Intact),group B(0 crosslink,NCL),group C(1 crosslink,1/2 of the rods,MCL),group D(1 crosslink,1/3 of the rods close to T14,PCL),group E(1 crosslink,2/3 of the rods away from T14,DCL)and group F(2 crosslinks,1/3 and 2/3 of the rods respectively,TCL). After all samples were subject to 10 000 times of fatigue test with biomechanics test machines,the axial compressive stiffness,range of the motion(ROM)of the 6 directions(flexion,extension,lateral bending,and axial rotation),and the maximum pullout of the screws of the T14 pedicle cortex perforation were measured and compared among these 6 groups. Results The axial compressive stiffness in groups A,C,D,E,and F was significantly higher than that in group B(all P<0.05),and group A was significantly higher than group F(P<0.05) . The maximum pullout in groups A,C,D,E,and F were significantly higher than that in group B(all P<0.05),and group A was significantly higher than group F and groups C,D,and E were significantly lower than group F(all P<0.05). ROM of flexion,extension,lateral bending,and axial rotation in groups A,B,C,D,E,and F were significantly lower than that in group B(P=0.000),and ROM of left and right axial rotation in groups C,D,and E were significantly higher than in that group F(P=0.000). Conclusions During the pedicle cortex perforation,adding of one crosslink can improve the stability of the pedicle cortex perforation,and adding of two crosslinks can approximately achieve the same stability as the pedicle screw fixation with no pedicle cortex perforation. The location of the crosslink has no obvious effect on the short segment of spinal fixation.

Objective To discuss perioperative rehabilitative care for flexor tendon injuries by examples of successful treatment of 30 cases of rupture of flexor tendon pollicis longus at ZoneⅡ.Methods From March 2001 to March 2006,30 cases of rupture of flexor tendon pollicis longus at ZoneⅡwere treated in our department by suture with double loops and crossing six strands before external fixation.Patients were encouraged to do rehabilitation exer- cises for functional recovery under the guidance and supervision of orthopaedists.Results All the patients were followed up for 6 to 16 weeks (mean,8 weeks).According to TAM (Total Active Motion) criteria,12 cases were rated as excellent,10 cases as good,6 cases as fair,and 2 cases as poor.The excellent to good rate was 73.3%.The 22 excellent and good cases could perform daily-life activities with affected fingers,such as writing,holding chopsticks and doing and undoing buttons.No patient experienced rerupture.Conclusion In perioperative rehabilitative care for rupture of flexor tendon pollicis longus at ZoneⅡ,it is important to have correct diagnosis,noninvasive and secure suture,safe external fixation and functional exercises under the guidance of orthopaedists.

Objective To discuss change the platelet counts by microscope rule by the platelet of Chinese people biological refer‐ence range in this laboratory blood analysis requirement .Methods Retrospectively analyzed according to the platelet of Chinese people biological reference range change the lab Sysmex XE‐5000 automatic blood cell analyzer platelet review rules .Change the platelet counts by microscope rules ,the differences between the microscopy rate and the microscopic examination of the platelet ag‐gregation rate .Results Before change the platelet counts by microscope rules the microscopy rate was 1 .94% .After change the platelet counts by microscope rules the microscopy rate was 3 .99% ,after change the platelet counts by microscope rules pushing blood piece rate increased significantly ,the difference was statistically significant(P<0 .05) .Conclusion According to the platelet of Chinese people biological reference range change the lab platelet review rules .We can increase the rate of microscopy ,and effec‐tively avoid the pseudothrombocytopenia and accurately guide clinical treatment .

BACKGROUND: Donor complications have been detected following autologous tendon transplantation for posterior cruciate ligament reconstruction. Although artificial tendon development and tissue-engineered tendon have achieved great progresses, there are some issues in clinical application. Since 1980's, allogenic tendon transplantation has aroused increasing attention. OBJECTIVE: To explore the selection of allogenic tendon materials and the effect of their application on reconstructing posterior cruciate ligament. METHODS: A total of 17 patients with posterior cruciate ligament injury of knee joint were treated with cryopreserved allogenic tendon by Tibial-inlay technique. During the operation, two tracts of tendons soaked in gentamicin saline for 15 minutes were conduplicated, and one end of the tendon was cancellous bone screw and fixed to the tibia attachment point of posterior cruciate ligament, and the other end was introduced into the joint through retention suture. The posterior joint capsule was repaired. The patient was placed at supine position, and the knee was flexed for 90°. The other end of the graft was introduced to femoral tunnel, and anterior drawer was tensed, and fixed by screw. RESULTS AND CONCLUSlN: The preoperative posterior drawer test of patients was >2+, including 7 cases of 3+ and 6 of 4+. The postoperative posterior drawer test was 0 in 4 cases, 1+ in 8 cases, 2+ in 4 cases and 3+ in 1 case, suggesting the posterior movement of the knee joint was significantly improved. Lysholm scores of patients were (48.5±4.3) points before operation and (88.3±5.4) points after operation. Results show that cryopreserved allogenic tendon by Tibial-inlay technique could restored function of posterior cruciate ligament with a favorable effect.

Objective To find an appropriate r3diobiological model for analyzing the biological effect of the radiotherapy for breast cancer by comparing different results computed by various types of radiobiological models. Methods DVHs database simulating breast conserving radiotherapy was set up,based on clinical DVHs data of the heart.the lung and PTV of 22 patients with early breast cancer given conventional tangential radiotherapy.Two models assessing NTCP of radiation pneumonitis and cardiac mortality and four models assessing TCP were compared by analyzing the effects of the parameters and DVH database input methods on the results. Results When mean irradiation dose of the whole lung was 30 Gy.the incidence of radiation pneumonitis was 32%and 54%predicted by NTCP-RSM model and NTCP-Lyman model,respectively.When 1%cardiac mortality of radiation was assumed,28 Gy and 40 Gy isodose should cover the heart assessed by the two models.The mean TCP were 21.1%.80.8%.38.4%and 41.0%when assessed by LQ-Poisson-TCP,Zaider-TCP,Poisson-TCP and Logit-TCP models,respectively.Setting various differential DVH(dDVH)bins had very few effect on the NTCP/TCP results in a certain model.Adopting physical dose or LQED2 affected the results with greater resu]ts for physical dose.Variation in α or β value,tumor cell density and D50 had significant effect upon TCP results in LQ-Poisson-TCP(P:0.000). Conclusions NTCP-Lyman model is better for predicting the incidence of radiation pneumonitis while NTCP-RSM model is better for predicting radiation-induced cardiac mortality.LQ-Poisson-TCP is the best TCP model for clinical application.Parameters selected for model can significantly affect the results.It is imporrant to understand the distinct characteristics of different models.