Ejaculatory Ducts ; Hemospermia ; Humans ; Male ; Seminal Vesicles

ObjectiveUsing Chromium-51 release assay, lactate dehydrogenase release assayand other methods to detect the cytotoxicity of CD19 CAR-T cells is cumbersome, with low repeatability and poor stability. This study aims to establish a label-free and real-time method for detectingspecific cytotoxicity of CD19 CAR-T cells.MethodsIn order to establish target cell models for cytotoxic assay of CD19 CAR-T cells by using Real Time Cellular Analysis (RTCA) system,the adherent human breast cancer cells were infected with lentiviral vectors encoding CD19. CD19 expression on the transduced cells was detected by flow cytometry. The cellsexpressing CD19 stably werethen sorted by fluorescence activated cell sorting (FACS).With such cells as target cells, CD19 CAR-T cells and BCMA CAR-T cells as effector cells, RTCAsystem was used to evaluate the cytotoxicity of CAR-T cells against target cells.ResultsMDA-MB-231 and SKBR3cells with stable expression CD19were obtained in this study.The results of flow cytometry showed that positive expression rate of CD19 in MDA-MB-231/CD19 cells and SKBR3/CD19 monoclonal cells were 99.03% and 98.91%,respectively.RTCA results showed that with MDA-MB-231 and MDA-MB-231/CD19 cells as target cells,CD19 CAR-T cells showed significant cytotoxicity to MDA-MB-231/CD19 cellsat the effector-target ratio of 5∶1, 1∶1 and 1∶5,but not to MDA-MB-231 cells. With SKBR3 and SKBR3/CD19 cells as target cells, CD19 CAR-T cells showed significant cytotoxicity to SKBR3/CD19 cellsat the effector-target ratio of 5∶1and 1∶1. When the effector-target ratio was 1∶5, there was no obvious cytotoxicity.The data of MDA-MB-231/CD19 or SKBR3/CD19 as target cells and CD19 CAR-T as effector cells were analyzed separately, showing that when the number of target cells was the same, the cytotoxicity detected by RTCA increased as the number of CD19 CAR-T cells increased.The cytotoxic assays of CD19 CAR-T cells showed specificity and dose-response relationship of CD19 CAR-T cytotoxicity against the target cells.ConclusionThis study established a method for evaluating cytotoxicity of CD19 CAR-T cells that is real-time, label-free, simple and convenient.

OBJECTIVE: To analyze the relationship between the distribution of lower limb alignment and short term clinical efficacy in patients with varus-type osteoarthritis after primary total knee arthroplasty (TKA). METHODS: From December 2016 to March 2018, 87 patients (101 knees) with knee osteoarthritis were treated with the first total knee arthroplasty by the same medical group, including 21 males(25 knees) and 66 females(76 knees), ranging in age from 51 to 85 years old, with a mean of (67.6±7.0) years old. According to the difference of hip knee ankle angle (HKA) after total knee arthroplasty, the patients were divided into 4 groups:neutral position group (group A), -3°≤HKA≤3°, 50 knees;slight varus group (group B), 3°< HKA<6°, 20 knees;severe varus group (group C), HKA≥6°, 20 knees;valgus group (group D), HKA<- 3°, 11 knees. The preoperative sex, age, body mass index, operative side, preoperative and postoperative knee joint activity, HSS score, KSS clinical and functional score were compared among the 4 groups, and the relationship between the force line distribution of femoral and tibial prosthesis and the recent clinical effect was compared. RESULTS: All the patients were followed up with a mean duration of(18.4±4.0) months. The range of motion of knee joint, HSS and KSS scores at the latest follow-up after operation in the 4 groups were higher than those before operation, and the difference was statistically significant (<0.001). There were significant differences in HSS and KSS scores among the 4 groups at the latest follow up (<0.05);and the results in group A were better than those in group C and group D (<0.05);the results in group B were better than those in group C and group D (< 0.05);there was no significant difference between group A and group B or group C and group D(>0.05). There was no significant difference in knee joint activity among the 4 groups. The score of femoral prosthesis force line within ±3°was better than that of the other group (<0.05), and the score of tibia prosthesis force line had no significant difference between within ±3° group and other group (>0.05). CONCLUSION The short term clinical efficacy of patients with knee varus osteoarthritis after primary total knee arthroplasty is related to the distribution of lower limbs alignment. The short-term clinical efficacy of slight inversion position can be similar to that of neutral position. The force line distribution of femoral prosthesis is related to the short term clinical efficacy after primary knee arthroplasty.
Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Knee ; Female ; Humans ; Knee ; Knee Joint ; Knee Prosthesis ; Lower Extremity ; Male ; Middle Aged ; Osteoarthritis, Knee ; Retrospective Studies

BACKGROUND: After total knee arthroplasty, some patients will receive the contralateral total knee arthroplasty again, but the influencing factors of contralateral knee surgery are not clear at present. OBJECTIVE: To evaluate the risk ratio of contralateral knee to total knee arthroplasty in patients with osteoarthritis of both knees after total knee arthroplasty, and to analyze the risk factors of contralateral total knee arthroplasty by clinical and radiographic indicators. METHODS: Data of 193 patients admitted to Affiliated Hospital of Xuzhou Medical University from 2013 to 2015 with bilateral knee osteoarthritis and undergoing unilateral total knee arthroplasty (follow-up time of 1-66 months) were retrospectively analyzed. The patients were divided into bilateral arthroplasty and unilateral arthroplasty groups based on whether the contralateral knee joint was replaced. K-M survival analysis was used to evaluate the risk ratio of contralateral knee total knee arthroplasty. Age, gender, body mass index, primary surgical side, non-operative knee Hospital for Special Surgery score, pain, mechanical axis perpendicular angle, medial and lateral joint space, femoral tibial angle, hip knee ankle angle, and Kellgren-Lawrence grade at initial surgery were used to assess the risk of contralateral total knee arthroplasty. RESULTS AND CONCLUSION: (1) Fifty-eight patients (30.1%) received contralateral total knee arthroplasty. The risk of nonoperative knee surgery at 1 year was 8%, and the risk of nonoperative knee surgery at 5 years was 32%. (2) The K-L grade was highly correlated with the risk of future nonoperative total knee arthroplasty (P < 0.001). The mean survival time of the nonoperative knee with K-L grade 4 was 45.7 months, and the 5-year surgical risk was 44.3%. (3) Age, gender, initial surgery side and body mass index, medial and lateral joint space, and hip knee ankle angle were not risk factors for progression to contralateral total knee arthroplasty. Nonoperative knee mechanical axis perpendicular angle (risk ratio=1.437, P < 0.001), femoral tibial angle (risk ratio=1.232, P <0.001) and K-L classification (risk ratio=2.110, P=0.005) were independent risk factors. Patients with femoral tibial angle ≥185.8° and mechanical axis perpendicular angle ≥2.75° had a higher risk of contralateral total knee arthroplasty (P <0.001). (4) The preoperative Hospital for Special Surgery score of the bilateral arthroplasty group was significantly lower than that of the unilateral arthroplasty group (P=0.039). Visual analogue scale pain score in the preoperative bilateral arthroplasty group was significantly higher than that in the unilateral arthroplasty group (P=0.013). At 6 months postoperatively, Hospital for Special Surgery score was improved in both groups, but still lower in the bilateral arthroplasty group than that in the unilateral arthroplasty group (P=0.003). (5) Not all patients with bilateral knee osteoarthritis need to receive contralateral total knee arthroplasty after unilateral total knee arthroplasty, and patients with severe knee osteoarthritis (K-L grade 4) are at high risk of needing contralateral total knee arthroplasty. Preoperative contralateral knee mechanical axis perpendicular angle, femoral tibial angle, Hospital for Special Surgery score and pain degree can be used as important factors to recommend patients or arrange a reoperation plan.

OBJECTIVETo observe the effect of admission blood urea and creatinine levels on mortality in elderly patients with hip fracture.

METHODSForm January 2013 to December 2014, 767 elder patients with hip fracture were treated in our hospital including 253 males and 514 females, aged from 65 to 96 years old with an average of(75.67±6.81) years old. According blood urea and creatinine levels, the 767 hip fracture patients were divided into four groups as follow: group A(blood urea>=5 mmol/L, creatinine>=70 μmol/L); group B (blood urea>=5 mmol/L, creatinine<70 μmol/L); group C (blood urea<5 mmol/L, creatinine>=70 μmol/L); group D(blood urea<5 mmol/L, creatinine<70 μmol/L). In group A, there were 211 patients including 70 males and 141 females, aged from 65 to 95 years old with an average of(80.24±6.51) years old; in group B, there were 355 patients including 125 males and 230 females, aged from 65 to 93 years old with an average of(78.46±7.09) years old; in group C, there were 36 patients including 11 males and 25 females, aged from 65 to 95 years old with an average of (77.83±6.78) years old; in group D, there were 165 patients including 47 males and 118 females, aged from 65 to 96 years old with an average of (76.71±8.35) years old. The survivals and dead patients in four groups were collected and in-hospital mortality rate, 3-month, 12-month and 18-month mortality rate of patients were calculated. COX regression analysis was performed on these data, and clinical significance of serum urea and creatinine at admission in the elderly patients was researched.

RESULTSAll 767 hip fracture patients were followed up from 18 to 24 months with an average of (21.33±1.25) months, 159 patients were died in follow up period. The in-hospital mortality rate in 3-month, 12-month and 18-month mortality rate of the patients with high blood urea and high blood creatinine (urea>=5 mmol/L, creatinine>=70 μmol/L) were 2.37%, 9.95%, 16.11% and 26.07%, and were higher than other three groups respectively. COX regression analysis revealed that the independent predictors effecting the mortality rate included age [=0.000, OR=1.375, 95%CI(1.155, 1.637)], blood urea at admission [=0.000, OR=1.375, 95%CI(1.155, 1.637)], and blood creatinine at admission[=0.037, OR=1.213, 95%CI(1.121, 1.484)].

CONCLUSIONSElderly hip fracture patients with high serum urea and high serum creatinine at admission indicate higher fatality rate. Age, serum urea and serum creatinine at admission were independent predictors of fatality rate of elderly hip fracture patients.

AIMTo examine the liver mechanism with which clenbuterol is explained how to affect growth metabolism.

METHODSTwenty-four adult SD rats were randomly divided into three groups, a control and two treatment groups. The technique of isolated perfused rat liver in vitro was used to study the effects of clenbuterol on urea nitrogen concentration of perfused medium, GPT activity and synthesis and secretion of IGF-I in isolated perfused rat liver.

RESULTSUrea-nitrogen concentration of perfused medium was significantly affected by clenbuterol in a dose-dependent and time-dependent manner. The urea-nitrogen level was decreased by 15.02% (P > 0.05),17.97% (P > 0.05), 26.76% (P < 0.05) and 30.08% (P < 0.01) for 1 h, 2 h, 3 h, 4 h after administering clenbuterol at the dose of 1 x 10(-6) mol/L, respectively, compared to that of control. 1 x 10(-8) mol/L CL had the similar effect on urea-nitrogen level. GTP activity of isolated perfused rat liver was inhibited by clenbuterol. The enzyme activity was decreased by 24.65% (P < 0.05) at the dose of 1 10(-6) mol/L CL in clenbuterol-treated 4h. The production and secretion of IGF-I were also influenced by clenbuterol in isolated perfused rat liver. IGF-I concentration of rat liver was increased by 19.77% (P < 0.05) in 4 h clenbuterol treatment (1 x 10(-6) mol/L). Meanwhile, IGF-I concentration of perfusion medium was also elevated though the difference was not significant compared with control.

CONCLUSIONIt is suggested that clenbuterol may improve growth metabolism by means of increasing nitrogen retention and enhancing IGF-I production and secretion of rat liver.

Animals ; Clenbuterol ; pharmacology ; In Vitro Techniques ; Insulin-Like Growth Factor I ; metabolism ; Liver ; drug effects ; metabolism ; Nitrogen ; metabolism ; Rats ; Rats, Sprague-Dawley

AIMTo examine the liver mechanism with which clenbuterol (CL) is explained how to affect growth metabolism.

METHODSThe technique of chronic poly catheter was used to study the effects of CL (0.8 mg/kg b w) on the hepatic flux of nitrogen, VFA and glucose in 4 sheep.

RESULTSThe urea-nitrogen flux in CL-treated period always was lower than that in control during 24 h. The average flux of urea-nitrogen in hepatic and portal vein were decreased by 16.86% (P < 0.01) and 15.51% (P < 0.05), respectively, compared with that of control. The peptide level in hepatic vein was decreased with the treatment of CL, average flux of peptide was decreased by 38.71% (P < 0.01). But the peptide level of portal vein in CL treatment period was similar to control. Moreover, VFA level in the portal vein was enhanced by CL, the average flux of acetate in portal vein was increased by 19.49% (P < 0.01). No difference of VFA level in hepatic vein was noted between CL-treated period and control. In addition, the glucose flux in hepatic vein was obviously increased with CL treatment, the average flux of glucose was increased by 25.96% (P < 0.01). And glucose flux in portal vein was also elevated during CL-treated period.

CONCLUSIONCL can affect growth metabolism of animal with increasing nitrogen deposition, improving absorption and utilization of VFA and enhancing glucose synthesis in sheep liver.

Animals ; Clenbuterol ; pharmacology ; Fatty Acids, Volatile ; metabolism ; Glucose ; metabolism ; Liver ; drug effects ; metabolism ; Sheep

OBJECTIVE: To explore the clinical significance of platelet closure time (PCT) in patients with multiple myeloma (MM). METHODS: Peripheral blood samples of 50 newly diagnosed MM patients treated in our hospital from July 2018 to November 2019 and 34 healthy persons underuent physical at the same time were collected. PCT induced by collagen/epinephrine (CEPI) and collagen/adenosinediphosphate (CADP) in peripheral blood were detected by PFA-200，and the clinical data included age, sex, leukocyte count, hemoglobin level, platelet count and level of serum creatinine, cystatin c, blood calcium, β-microglobulin (β-MG), bone marrow plasma cells, light chain protein, as well as the MM types, ISS stage of patients were collected. RESULTS: The level of PCT in MM patients was significantly higher than that in healthy persons; the level of PCT were significantly increased with the increasing of ISS stage in newly diagnosed MM patients; After chemotherapy with bortezomib/dexamethasone (BD), the level of PCT in 15 patients who were responded to the treatment was significantly lower than those before treatment. CONCLUSION The platelet closure time is abnormal in MM patients, moreover, relates to the progress of the disease. It has an important clinical significance for the evaluation of diagnostic stage and therapeutic efficacy evaluation of MM patients.
Blood Platelets ; Bone Marrow ; Bortezomib ; Humans ; Multiple Myeloma ; Platelet Count

The present study was aimed to investigate the role of GluN2B-BDNF pathway in the cerebrospinal fluid-contacting nucleus (CSF-CN) in neuropathic pain. Intra-lateral ventricle injection of cholera toxin subunit B conjugated with horseradish peroxidase (CBHRP) was used to label the CSF-CN. Double-labeled immunofluorescent staining and Western blot were used to observe the expression of GluN2B and BDNF in the CSF-CN. Chronic constriction injury of sciatic nerve (CCI) rat model was used to duplicate the neuropathic pain. Pain behavior was scored to determine the analgesic effects of GluN2B antagonist Ro 25-6981 and BDNF neutralizing antibody on CCI rats. GluN2B and BDNF were expressed in the CSF-CN and their expression was up-regulated in CCI rats. Intra-lateral ventricle injection of GluN2B antagonist Ro 25-6981 or BDNF neutralizing antibody notably alleviated thermal hyperalgesia and mechanical allodynia in CCI rats. Moreover, the increased expression of BDNF protein in CCI rats was reversed by intra-lateral ventricle injection of Ro 25-6981. These results suggest that GluN2B and BDNF are expressed in the CSF-CN and alteration of GluN2B-BDNF pathway in the CSF-CN is involved in the modulation of the peripheral neuropathic pain.
Animals ; Brain-Derived Neurotrophic Factor ; Hyperalgesia ; Neuralgia ; Rats ; Rats, Sprague-Dawley ; Sciatic Nerve

Objective: To summarize the effects of medial upper arm bilobed free flaps in the repair of two skin and soft tissue defects of hand. Methods: From May 2014 to May 2016, 7 patients of two skin and soft tissue defects of hand with exposures of phalanges and tendons were treated in Xuzhou Renci Hospital, including 5 males and 2 females, aged from 19 to 41 years. Each defect area ranged from 3.0 cm×1.5 cm to 6.0 cm×3.0 cm. The medial upper arm bilobed free flaps were used to repair the defects, and the area of each lobe of the flaps ranged from 4.0 cm×2.0 cm to 8.0 cm×3.5 cm. The donor sites were sutured directly. The survival of the flaps was recorded. During follow-up, the recovery of donor and recipient sites was observed, the two-point discrimination distance of the flaps was measured, and the hand function was assessed by the trial criteria of the upper limb functional assessment of the Hand Surgery Society of the Chinese Medical Association. Results: All flaps survived smoothly, and the wounds and donor site incisions were healed. All patients were followed up, and the follow-up time lasted for 6 to 15 months. The color and texture of the flaps were similar to the surrounding normal skin, and the shape of the flaps was good. There was no obvious scar in the donor site, and the elbow joint function was normal. One patient developed ulnar numbness one month after operation and relieved after 3 months of treatment with neurotrophic drugs and local physiotherapy, etc. Six months after operation, the two-point discrimination distance of the flaps was 5.5-8.0 mm, and the hand function evaluation was excellent in 3 cases, good in 3 cases, and middle in 1 case. Conclusions The medial upper arm bilobed free flap has both good texture and good appearance, and the scar of donor site is concealed. It is a good method to repair two skin and soft tissue defects of hand.