Objective To investigate the assembl ing and cl inical appl ication of the video output system util izing teaching sight glass of surgical microscope. Methods Between June 2009 and April 2010, 10 patients with craniocervical junction malformation were treated by the method of transoral-transpharyngeal approach with the microscope and videooutput system under the direct vision. There were 6 males and 4 females with an average age of 32 years (range, 13-52 years). Three cases had the history of injury and 7 cases had no history of definite injury. The disease duration was from 10 months to 12 years (median, 5 years). The main cl inical symptoms were brevicoll is or torticoll is; 2 patients had malformation appearance and 4 patients had occi put-cervical pain. The physical examination showed that all patients had the symptoms that upper cervical cord was damaged; the imaging examination showed that all patients had basilar invagination, atlantoaxial dislocation, and ossification. Before and after operations, the functions of nerve were evaluated by Japanese Orthopaedic Association (JOA) scoring, the improvement rate was calculated to evaluate the efficacy. Results By the video output system assembly, 15.1 mill ion pixels high-definition images could be collected and reached 1 920 × 1 080 pixels video camera, so assistants or medical students could watch the cl inical operation directly. All patients had no neural injury or cerebrospinal fluid leakage during operation. Basilar invagination and atlantoaxial dislocation were corrected. Infection at incision occurred in 1 patient; other incisions healed by first intention without early compl ication. All patients were followed up 6-16 months (mean, 13.5 months). The average JOA score was increased from 10.2 preoperatively to 15.5 at 6 months postoperatively with an improvement rate of 77.9%. At 12 months after operation, bony fusions were achieved. Conclusion The miscroscope and video output system can improve the effectiveness of the original surgical microscope. It makes visual fields much clearer and operations more accuratewith a few compl ications.
Objective To study the effect of substance P ( SP) on int racellular f ree calcium concent ration in human poorly-differentiated gast ric cancer cell in vitro. Methods Human gast ric cancer cell line MKN45 was cultured in RPMI 1640. Then the cells were loaded with specific calcium fluorescent probe Furu23/ AM. ASN21377642 (NK21 receptor antagonist) , Nicardipine (calcium channel blocker) and different concent rations of SP were used to treat gast ric cancer cells. The concent ration changes of int racellular free calcium were detected by laser scanning confocal microscope. Results It was found that 10 , 50 and 100 nmol/ L SP could significantly increase the int racellular free calcium concent ration of gast ric cancer cells in Hanks solutions , which contain ext racellular calcium ( P lt;0. 05) , and the change was in a dose-dependent manner ( P lt; 0. 05) . When there was ext racellular calcium existed ,the increasing amplitude of intracellular f ree calcium concent ration was significantly higher than that when there was no extracellular calcium ( Plt; 0. 05) . And when Hanks solutions were pretreated with ASN21377642 and Nicardipine , the effects of 100 nmol/ L SP were partly inhibited , and the concent rations of int racellular f ree calcium were significantly lower than those in group s without pret reatment s ( P lt; 0. 05) . Conclusion SP can significantly increase free calcium concent ration in the gastric cancer cells. Releasing of stored calcium in the cells and influx of extracelluar calcium may contribute to the elevation of int racellular free calcium concentration.
ObjectiveTo design a method for observing pulmonary microcirculation in vivo in a native tissue environment, due to the high incidence of lung diseases and the advantages of animal experiments in vivo.MethodsTen BALB/c male mice were randomly divided into group A and group B, with five mice in each group. A self-made apparatus was used to control the movement towards local lung tissues in order to get a stabilized observation plane, and then a 5-minute video was shot with laser confocal scanning microscope. During the filming, the mice in group A were injected with fluorescein isothiocyanate-dextran via the tail vein, and the mice in group B were injected with green fluorescent protein-platelets (extracted from the blood of tie2-cre&rosa26-tomato-EGFP transgenic black C57 male mice). The data of group A was used to observe the lungs perfusion and the damage to tissue by this method, and the data of group B was used to observe the movement of platelets.ResultsImage of lung structure obtained by this method was clear and stable. Mean areas of alveolus in an instant and at the 30th, 60th, 120th, 180th, and 300th second were (1 603±181), (1 588±183), (1 528±363), (1 506±353), (1 437±369), (1 549±307) μm2, respectively, and there were no significant differences between each time point (P>0.05). The video was smooth, the rapid movement of platelets was recorded and the particles were clear and without tailing; after the observation, hematoxylin-eosin staining showed no obvious damage to the lung tissue.ConclusionThe method can be used for the observation and research of the lung microcirculatory system in the living state of the mouse, and provides a methodological basis for studies of other lung diseases in vivo.
Objective To study the influence of the immersed time by hydrogen dioxide on the characteristics of bovine cancellous bone granules in various periods. Methods Ten 24-month-old Qinchuan bovine, male or female, weighing 150-170 g, were selected. Cancellous bone granules from metaphysic of bovine long bone were cut into cubes of 5 mm × 5 mm ×5 mm and immersed by 8.8 mol/L hydrogen dioxide for 0, 12, 24, 36, 48, 60 and 72 hours separately. Determination of ash, scanning electron microscope, X-ray energy spectrum and micro CT were used to investigate the changes of composition, structure and qual ity of bone. Results With the immersed time increasing, the contents of organics in the bone cancellous were reduced gradually, and obviously decreased during the periods of 0 to 24 hours and 60 to 72 hours (P lt; 0.05). The contents of calcium and phosphorus decreased gradually, they could not be detected almost after 60 days (P lt; 0.05). Bone mineral density and bone mineral content were decreased obviously after 60 hours (P lt; 0.05). The bone trabecula became sl immer and trabecular spacing became larger. Conclusion Hydrogen dioxide can be used to remove the antigen in xenogeneic bone; however as the time increasing (more than 60 hours) the composition and structure will be damaged. Thus it is important to control the immersed time for maintaining the biological characteristics of xenogeneic bone substitute as well as el iminating antigen by hydrogen dioxide.
OBJECTIVE: To investigate the expression and distribution of platelet derived growth factor receptor-beta(PDGFR-beta) in normal skin and keloid and to discuss its biological function in keloid formation. METHODS: 1. To detect the expression and distribution of PDGFR-beta in normal skin and keloid tissue by immunohistochemistry; 2. To detect the receptor expression in vitro by Flow cytometry (FCM); 3. To detect the subcellular distribution of receptor by Laser confocal microscope. RESULTS: 1. Immunohistochemistry showed that normal skin and keloid tissue were almost the same in expression but different in distribution of PDGFR-beta; 2. There was more expression of PDGFR-beta in normal fibroblasts than that in keloid fibroblasts in vitro by FCM; 3. Laser confocal microscope revealed that the PDGFR-beta concentrated on the surface of cell membrane in keloid fibroblasts, but in normal skin fibroblasts, the receptors were coagulated on the nuclear membrane and intranucleus. CONCLUSION: Compared with the fibroblasts in vivo, there was a difference of the PDGFR-beta expression in fibroblasts in vitro, more expression of PDGFR-beta in normal fibroblast than that in keloid fibroblast in vitro; and the subcellular distribution of PDGFR-beta was different in normal skin and keloid fibroblasts. The characteristics of the expression and distribution of PDGFR-beta in keloid may contribute to the formation of keloid.
Objective To explore the feasibility of microscopic spermatic vein ligation for unilateral varicocele under daytime surgical mode. Methods The clinical data of patients with unilateral varicocele who underwent daytime surgical treatment at the General Hospital of Northen Theater Command between January and December 2022 were retrospectively analyzed. The relevant indicators of patients were collected, including age, surgical time, intraoperative blood loss, total hospitalization expenses, and postoperative condition. Results A total of 159 patients were included, aged 18-41 years, with an average age of (27.79±4.80) years, an average surgical time of (49.23±5.57) minutes, and an average intraoperative blood loss of (8.94±3.58) mL. One patient had fever, nausea and acid regurgitation on the 2nd day after discharge, and one patient had urinary retention on the day after operation. Five patients had a pain score of ≥3 in the surgical area on the 1st day after surgery. All patients had no postoperative incisions with bleeding or hematoma. Follow up showed that all patients returned to normal life on the 3rd day after surgery and returned to work on the 10th day after surgery. At 3-6 months after surgery, the sperm quality of 94 patients improved. One patient experienced disease recurrence 6 months after surgery. Conclusion Microscopic spermatic vein ligation for unilateral varicocele under daytime surgical mode is safe and feasible, and is worthy of promotion.
Objective To observe the ultrasonographic features of idiopathic uveal effusion syndrome (IUES). MethodsA retrospective controlled study. From January 2012 to December 2023, 13 patients with IUES (26 eyes) diagnosed by ophthalmology examination in Department of Ophthalmonogy of Beijing Tongren Hospital (IUES group) and 22 healthy people with 30 eyes (control group) were included in the study. Both eyes of all participants were examined by color Doppler ultrasound (CDU) and ultrasonic biomicroscopy (UBM). The thickness of the ocular wall at 300 μm on the temporal side of the optic disc was measured by CDU. UBM was used to measure the thickness of the nasal and temporal scleral processes. Corneal thickness (CT), anterior chamber depth (AD), lens thickness (LT) and axial length (AL) were measured by A-mode ultrasound. There were no significant differences in age (t=0.842), sex component ratio (χ2=0.540), eye difference (χ2=0.108) and AL (t=0.831) between IUES group and control group (P＞0.05). The CDU and UBM imaging features and biometrics of IUES affected eyes were observed. Independent sample t test was used for comparison between groups. ResultsCDU examination results showed that in 26 eyes of IUES group, choroidal detachment occurred in 20 eyes (76.9%, 20/26), which showed arc-shaped band echo connected with peripheral and equatorial eye wall echo, with uniform low echo area below, and blood flow signal could be seen on the band echo. The echo thickened and decreased in 4 eyes (15.3%, 4/26). Nine eyes (33.3%, 9/26) were accompanied by retinal detachment, which showed that the posterior pole vitreous echo was connected to the optic disc echo, and the blood flow signal was seen on the ribbon echo. UBM results showed ciliary detachment in 22 eyes (84.7%, 22/26), showing a spongy thickening of the ciliary body with interlamellar echo separation and an echoless area between the sclera. Ciliary body echo thickened and decreased in 2 eyes (7.7%, 2/26). Shallow space between ciliary body and sclera was observed in 2 eyes (7.7%, 2/26). Compared with the control group, CT (Z=2.054), LT (Z=1.867), scleral thickness (Z=2.536) and ocular wall thickness (Z=2.094) were thickened in IUES group, and AD (Z=1.888) were decreased, with statistical significance (P＜0.05). ConclusionsThe CDU of IUES is characterized by a thickened echo of the ocular wall and a uniform low echo area under the detached choroid. UBM is characterized by a spongy thickening of the ciliary body echo with interlaminar echo separation.
ObjectiveTo investigate the effectiveness of posterior microscopic mini-open technique (MOT) decompression in patients with severe spinal canal stenosis resulting from thoracolumbar burst fractures.MethodsThe clinical data of 28 patients with severe spinal canal stenosis caused by thoracolumbar burst fractures, who were treated by posterior microscopic MOT, which performed unilateral or bilateral laminectomy, poking reduction, intervertebral bone graft via spinal canal, and percutaneous pedicle screw fixation between January 2014 and January 2016 were retrospectively analyzed. There were 21 males and 7 females with a mean age of 42.1 years (range, 16-61 years). The involved segments included T11 in 1 case, T12 in 4 cases, L1 in 14 cases, and L2 in 9 cases. According to AO classification, there were 19 cases of type A3, 9 of type A4. According to American Spinal Injury Association (ASIA) grading, 12 cases were grade C, 13 grade D, and 3 grade E. The time between injury and operation was 3-7 days (mean, 3.6 days). To evaluate effectiveness, the changes in the visual analogue scale (VAS), percentage of anterior height of injured vertebrae, Cobb angle, rate of spinal compromise (RSC), and ASIA grading were analyzed.ResultsAll patients were performed procedures successfully. The operation time was 135-323 minutes (mean, 216.4 minutes). The intraoperative blood loss was 80-800 mL (mean, 197.7 mL). The hospitalization time was 10-25 days (mean, 12.5 days). The incisions healed primarily, without wound infection, cerebrospinal fluid leakage, or other early complications. All the 28 patients were followed up 12-24 months (mean, 16.5 months). No breakage or loosening of internal fixation occurred. All fractures healed, and the healing time was 3-12 months (mean, 6.5 months). Compared with preoperative ones, the percentage of anterior height of injured vertebrae, Cobb angle, and RSC at immediate after operation and at last follow-up and the VAS scores at 1 day after operation and at last-follow were significantly improved (P<0.05). There was no significant difference in the percentage of anterior height of injured vertebrae and Cobb angle between at immediate after operation and at last follow-up (P>0.05). But the RSC at immediate after operation and VSA score at 1 day after operation were significantly improved when compared with those at last follow-up (P<0.05). The ASIA grading at last follow-up was 1 case of grade C, 14 grade D, and 13 grade E, which was significantly improved when compared with preoperative ones (Z=3.860, P=0.000).ConclusionMOT is an effective and minimal invasive treatment for thoracolumbar AO type A3 and A4 burst fractures with severe spinal canal stenosis, and it is beneficial to early rehabilitation for patients.
Objective To comprehend the pathological features and possible pathogenesis of avascular necrosis of the femoral head (ANFH) by morphology and immunohistochemical observation of osterix (OSX) and adiponectin through in vitro traumatic and non-traumatic ANFH specimens, so as to provide a theoretical basis for cl inical treatment. Methods Sixty-six ANFH specimens were collected from 66 cl inical cases undergoing hip replacement surgery. Twenty-four cases of traumatic ANFH (group A) included 17 males and 7 females, aged 21 to 70 years with an average of 56.5 years; 23 cases of steroid-induced ANFH (group B) included 16 males and 7 females, aged 56 to 72 years with an average of 61 years; and 19 cases of alcohol ic ANFH (group C) were males, aged 55 to 67 years with an average of 58.5 years. Bone tissue was got from weight-bearing and non-weight-bearing area of the femoral head respectively. The basic pathological changes was observed by HE staining under the optical microscope, and the percentage of empty bone lacuna and the percentage of trabecular bone area were calculated. The morphological changes of ANFH in different groups were observedby scanning electron microscope (SEM). OSX and adiponectin expression were detected by immunohistochemical technique. Results Gross of the femoral head surface in each group was rough, collapse, articular cartilage loss, osteophyte formation; cross section: dark red in group A, and yellow in groups B and C. HE staining showed that weight-bearing area of ANFH have similar morphological features in three groups. In non-weight-bearing area of groups B and C, the fat cells in bone marrow markedly increased and were hypertrophic; however there were more fibrous tissue in group A. There were statistically significant differences (P lt; 0.001) in the percentage of empty bone lacuna of the weight-bearing and non-weight-bearing area among three groups. There were no statistically significant differences (P gt; 0.05) in the percentage of trabecular bone area among three groups. The SEM observation showed that three groups had similar pathological changes. Brown granules for OSX and adiponectin positive substance were mainly located in the osteoblast of bone marrow of the femoral head. There was statistically significant difference (P lt; 0.05) in the average absorbency (A) value of OSX between group A and groups B, C, but there was no statistically significant difference (P gt; 0.05) between groups B and C. While there was no statistically significant difference (P gt; 0.05) in the A value of adiponectin among three groups. Conclusion Hormones and alcohol necrosis have more obviously fatty degeneration, but the repair capacity of traumatic femoral head necrosis is ber than that of hormones and alcohol necrosis. Alcohol and hormones have inhibitory action on the OSX-mediated osteogenic differentiation. Hormones and alcohol may not affect osteoblast expressing adiponectin and its receptors.
The development of intravital microscopy (IVM) has enabled researchers to perform in-situ, real-time observations of pulmonary micro-circulation at the cellular level, and has become instrumental for researching the immune micro-environment of pulmonary diseases. This article introduces the developments in constructing the pulmonary imaging window and summarizes the current light microscopy techniques used for lung IVM with regard to its relevant functional and application features, which includes wide field fluorescence microscopy, confocal microscopy, as well as two-photon microscopy. It then provides examples of IVM of pulmonary immune response in inflammation and infection in murine models, and finally specifies the technological limitations to provide reference for researchers to systematically learn and understand the technology.