Objective To study the feasibility of using morphological changes of bronchioles and lymph nodes in rats for estimating the postmortem interval(PMI).Methods Thirty-five 8-week-old male SD rats were sacrificed by cervical dislocation and placed in a constant temperature and humidity box.Lung tissues of 5 rats were extracted at each time point within 24 hours after death for routine HE and PHH3 immunohistochemical staining.Image J was used to measure mucosal area,extramucosal area,luminal area of small bronchi,Then calculate the ratio of mucosal area to luminal area,extramucosal area to extramucosal,extramucosal area to mucosal area.imageJIHCprofiler was used to measure the positive degree of PHH3 immunohistochemical staining in lymph node nuclei,and the above calculation results and the positive degree of PHH3 staining were statistically analyzed by GraphPad Prism 9.5.Results The ratio of mucosal area to lumen area showed an overall downward trend with the increase of PMI(0h was statistically different compared with 18h and 24h,P<0.05).The ratio of extramucosal to lumen area and extramucosal to mucosa area showed an upward trend with the increase of PMI(0h was statistically different from other time points,P<0.05),the positive degree of PHH3 immunohistochemical staining was significantly decreased at 24 h after death(P<0.05).Conclusion The morphological changes of pulmonary bronchiole and lymph nodes showed a certain regularity within 24h after death,which was helpful to infer the early PMI,lymph node nucleus PHH3 was expected to be a critical index to distinguish early and late PMI.

Objective To study the feasibility of using morphological changes of bronchioles and lymph nodes in rats for estimating the postmortem interval(PMI).Methods Thirty-five 8-week-old male SD rats were sacrificed by cervical dislocation and placed in a constant temperature and humidity box.Lung tissues of 5 rats were extracted at each time point within 24 hours after death for routine HE and PHH3 immunohistochemical staining.Image J was used to measure mucosal area,extramucosal area,luminal area of small bronchi,Then calculate the ratio of mucosal area to luminal area,extramucosal area to extramucosal,extramucosal area to mucosal area.imageJIHCprofiler was used to measure the positive degree of PHH3 immunohistochemical staining in lymph node nuclei,and the above calculation results and the positive degree of PHH3 staining were statistically analyzed by GraphPad Prism 9.5.Results The ratio of mucosal area to lumen area showed an overall downward trend with the increase of PMI(0h was statistically different compared with 18h and 24h,P<0.05).The ratio of extramucosal to lumen area and extramucosal to mucosa area showed an upward trend with the increase of PMI(0h was statistically different from other time points,P<0.05),the positive degree of PHH3 immunohistochemical staining was significantly decreased at 24 h after death(P<0.05).Conclusion The morphological changes of pulmonary bronchiole and lymph nodes showed a certain regularity within 24h after death,which was helpful to infer the early PMI,lymph node nucleus PHH3 was expected to be a critical index to distinguish early and late PMI.

Objective:To investigate the effects of high-energy shock and vibration on cortex injury and peripheral blood immune cells in goats.Methods:Seventeen Boer goats without gender preference were selected. By using random number tables, the goats were divided into normal control group ( n=5) and shock and vibration injury group ( n=12). The goats in the normal control group were anatomized routinely and their brain was collected after being sacrificed without any other treatment. The goats in the high-energy shock and vibration model group were placed on a loading table (part of the BY10-100 instant shock and vibration simulation platform) in a restrained state, and made into a high-energy shock and vibration injury model induced by a vertical impact waveform generator. The intravenous blood samples were taken from the goats in the shock and vibration injury group before and at 0, 3, 6 and 24 hours after injury.Then, the goats were sacrificed and the following procedures were the same as the normal control group. At 24 hours after injury, the brain injury and the histopathological changes of the cerebral cortex in the normal control group and shock and vibration injury group were observed by gross pathological and anatomical examination and HE staining. The mRNA expression of zonula occludens 1 (ZO-1), tight junction protein 5 (Claudin-5), glial fibrillary acidic protein (GFAP), ionized calcium binding adaptor molecule 1 (IBA-1), interleukin (IL)-1β, IL-6 and cluster of differentiation antigen 177 (CD177) of the cerebral cortex in the normal control group and shock and vibration injury group were measured through fluorogenic quantitative polymerase chain reaction. The expression of ZO-1 and Claudin-5 proteins of the cerebral cortex in the normal control group and shock and vibration injury group were detected by Western blotting. Hematology analyzer and coagulation analyzer were used to detect white blood cell count, neutrocyte, lymphocyte, monocyte, prothrombin time 1 (PT-1), prothrombin time international normalized ratio (PT-INR), activated partial thromboplastin time (APTT), thrombin time (TT), prothrombin activity (PTA) and fibrinogen (FIB) levels in goats of the shock and vibration injury group before and at 0, 3, 6 and 24 hours after injury, respectively. Results:At 24 hours after injury, no visible contusion or necrosis was found in goat brain tissue in the shock and vibration injury group; the cerebral micro-vessels presented with a local dilation, hyperemia, edema, aggregation of inflammatory cells, disruption of vessel walls and leakage of red blood cells. These changes were not observed in the normal control group. In the shock and vibration injury group, ZO-1 and Claudin-5 mRNA expressions in the cerebral cortex were 0.25±0.10 and 0.09(0.04, 0.44) respectively, which were significantly lower than those of the normal control group [1.00±0.15 and 0.99(0.80, 1.20)]; GFAP, IBA-1, IL-1β, IL-6 and CD177 mRNA expression levels were 4.40(3.88, 6.75), 2.60±1.07, 3.04±0.51, 2.71±0.45 and 2.93±0.62 respectively, which were significantly higher than those of the normal control group [1.00(0.78, 1.22), 1.00±0.37, 1.00±0.27, 1.00±0.57 and 1.00±0.35]; ZO-1 and Claudin-5 protein expression levels were 0.41±0.06 and 0.42±0.11 respectively, which were significantly lower than those of the normal control group (1.08±0.12 and 0.91±0.23) (all P<0.01). In the shock and vibration injury group, the levels of white blood count, neutrocyte, and lymphocyte in peripheral blood were (13.7±3.3)×10 9/L, (35.3±14.8)% and (57.2±15.1)% respectively before injury, (19.4±3.1)×10 9/L, (60.5±12.5)% and (33.6±14.2)% respectively at 3 hours after injury, and (20.6±3.6)×10 9/L, (63.6±13.0)% and (30.9±15.0)% respectively at 6 hours after injury. By contrast, the levels of white blood count and neutrocyte were significantly increased but the level of lymphocyte was significantly decreased at 3 and 6 hours after injury ( P<0.05 or 0.01); the levels of the above indicators showed no significant changes at 0 and 24 hours after injury (all P>0.05); the level of monocyte did not change significantly at all time points before and after injury (all P>0.05). The levels of PT-1, PT-INR, APTT, TT, PTA and FIB in the shock and vibration injury group did not change significantly at each time point before and after injury (all P>0.05). Conclusion:Cerebral cortex microvascular injury and disruption of blood-brain barrier can be initiated in the early stage of high-energy shock and vibration injury in goats, accompanied by the presence of central and peripheral inflammatory response.

OBJECTIVE: To explore the genetic basis for a neonate featuring developmental delay. METHODS: Clinical examination and laboratory tests were carried out for the patient. Peripheral venous blood samples of the proband and his parents were extracted and subjected to target capture next generation sequencing. Candidate variant was verified by Sanger sequencing. RESULTS: The patient, a four-month-old male, has presented with developmental delay and weakness of limbs. Genetic testing revealed that he had harbored a novel c.1432C>T variant of the TNPO3 gene, which was inherited from his mother. The nonsense variant has resulted in premature termination of protein translation and was predicted to be pathogenic by bioinformatics analysis. CONCLUSION The heterozygous c.1432C>T variant of the TNPO3 gene probably underlay the limb-girdle muscular dystrophies form 1F in this patient. Above finding has enriched the variation spectrum of the TNPO3 gene.
Genetic Testing ; Heterozygote ; High-Throughput Nucleotide Sequencing ; Humans ; Infant ; Male ; Muscular Dystrophies, Limb-Girdle/genetics* ; Mutation ; Phenotype ; beta Karyopherins/genetics*

Objective To establish the model of rabbit vena arterialization, so as to investigate the difference of mechanical parameters between arteries and veins as well as before and after arterialization. Methods Twenty-four rabbits were randomly divided into experimental group (n=12) and control group (n=12). By establishing the rabbit vena arterialization model for experimental group, the arterial blood could flow into the veins. After model creation, the vein would be removed 4 weeks after surgery. In the meantime, the external jugular veins and cephalic arteries extracted from control group were acquired. Compressive pressurizing and stretching tests on all vessels were conducted at the same time(including arteries, veins and arterialized veins). Observation was supported by HE staining and immune tissue chemical techniques. Results There were no deaths among the 24 rabbits, with unobstructed blood flow in veins. With the increase of intravascular pressure, the outer diameter of veins changed at first and then stabilized at a fixed value. The elasticity of veins was worse than that of arteries. The external diameter of veins increased rapidly with internal pressure of veins increasing and reached its extreme elasticity. Comparatively, the elasticity of arteries increased slowly. HE staining results showed that thickness of the vascular wall was thinner, while it became thicker after vena arterialization. After vena arterialization, proliferating cell nuclear antigen（PCNA） and α-actin showed positive results. It further proved that proliferation existed among smooth muscle cells, and veins showed the tendency of restenosis again. The elasticity of veins after transplantation into the arterial system was improved compared with that before transplantation. Conclusions Accompanied by the increasing pressure, the vein could reach its elasticity extremity faster than the artery. Under such a long-term high pressure, vein intima was vulnerable. After vena arterialization, with the gradual thickening of vein intima, the tendency of vessel restenosis was obvious, and the elasticity of veins has been improved after transplantation.

Objective To evaluate the effect of regional hypothermia on limb ischemia-reperfusion (I/R) injury in a rabbit model of hemorrhagic shock.Methods Eighteen healthy adult New Zealand white rabbits of both sexes,weighing 2.0-2.5 kg,were divided into 3 groups (n=6 each) using a random number table:sham operation group (S group),limb I/R group (I/R group) and regional hypothermia group (RH group).Rabbits were shot with 0.25 g steel ball with a 7.62 mm smoothbore at the middle and lower 1/3 of the left hindlimb in I/R and RH groups.When the blood pressure was stable for 30 min after injury,blood was drawn via the right femoral artery at a rate of 2 ml/min until mean arterial pressure decreased to 45-50 mmHg.Then a rubber tourniquet was used to ligate left hindlimbs 10 min later.The injured limb was cooled down for 4 h after applying tourniquet to maintain the subcutaneous temperature at 9-11 ℃ in group RH.After tourniquet was applied for 4 h,perfusion was restored and lasted 6 h.After catheterization (T0) and at 2,4 and 6 h of reperfusion (T1-3),blood samples were collected from the right femoral artery for blood gas analysis and for determination of tumor necrosis factor-α (TNF-α),interleukin-2 (IL-2) and IL-6 concentrations in serum (by enzyme-linked immunosorbent assay).The animals were sacrificed at T3,and tissues around the margin of the inferior lobe of right lung were obtained for examination of the pathological changes which were scored and for determination of wet to dry weight ratio (W/D ratio).Results Compared with group S,the respiratory rate was significantly increased and PaCO2 was decreased at T2,3,the pathological score and W/D ratio were increased,and the concentrations of TNF-α,IL-2 and IL-6 in serum were increased at T1-3 in I/R and RH groups (P＜0.05).Compared with group I/R,the respiratory rate was significantly decreased and PaCO2 was increased at T2,3,the pathological score and W/D ratio were decreased,and the concentrations of TNF-α,IL-2 and IL-6 in serum were decreased at T1-3 (P＜0.05),and the pathological score was significantly attenuated in group RH.Conclusion Regional hypothermia can reduce limb I/R injury in a rabbit model of hemorrhagic shock,and the mechanism may be related to inhibiting systemic inflammatory responses.

ObjectiveTo evaluate the effect of high-frequency jec ventilation and bilevel positive airway pressure (BiPAP) ventilation in the severe dyspnea caused by central airway stenosis and explore the deferent application of two ventilation models.MethodsA retrospective analysis was designed to collect the serious dyspnea patients caused by central airway stenosis from January 2006 to January 2009.The patients were divided into group H and group B according to the different ventilation models,and the effect of relieving dyspnea and ameliorating hypoxemia was compared.ResultsSeven cases were in group H,and 9 cases were in group B.The therapeutic effect of relieving dyspnea was 14.29 ％(1/7) in group H.There were only 4 cases accepting high-frequency jec ventilation and the effect of relieving dyspnea was 25.00％(1/4).Two cases failed in high-frequency jec ventilation treatment and succeeded in BiPAP treatment thereafter,and 1 case failed in BiPAP treatment initially but was treated effectively by high-frequency jec ventilation after dyspnea relapsed.In group B,there were only 6 cases accepting BiPAP ventilation and the effect ofrelieving dyspnea was 83.33％ (5/6),and after adding the cases who accepting the sequential therapy of high-frequency jec ventilation and BiPAP ventilation the effect rate was 88.89％ (8/9).The difference had statistical significance(P =0.006).In ameliorating hypoxemia,the effect rate of group H was 100.00％ (7/7),of group B was 88.89％(8/9),and the difference had no statistical significance (P =0.563 ).Conclusions The high-frequency je ventilation is effective to relieve hypoxemia but is short of relieving dyspnea and subjective symptom,and it is suit for life support preoperative and intraoperative.The BiPAP ventilation is effective to relieve dyspnea and subjective symptom and hypoxemia,so the treatment effect is better than high-frequency jec ventilation,and it is suit in relieving symptom temporarily,pre-operative preparation and malignant tumor palliative treatment.

Objective To evaluate the regional cerebral metabolic changes in different episodes by magnetic resonance spectroscopy (MRS) after explosive brain injury in rabbits. Methods Fortyfive New Zealand white rabbits were randomly divided into eight groups, ie, normal control group( 10 rabbits) and trauma group (35 rabbits). The explosive injury in trauma group was induced by explosion of 600 mg TNT equivalent of paper detonators at 6.5 cm above the rabbit brain. The rabbits in trauma group was divided into 1,6, 12, 24 hours, 3, 7, 14 days subgroups (6 rabbits per group). The survival rate was observed at different time points after explosive injury. The MRS was used to detect the regional cerebral metabolic changes including N-acetylaspartate (NAA)/creatine (Cr) ratio and choline(Cho)/Cr ratio as well as evolution of blast injuries over time. Results The rabbits survived for overseven days in the trauma groups, with typical brain contusion manifested by pathological and conventional MRI. Compared with the normal control group, the NAA/Cr ratio was markedly decreased at one hour after injury, slightly rose again at 24 hours and fell again after seven days. The Cho/Cr ratio was markedly increased at one hour after injury, slightly fell again at 12 hours and rose again at three days after injury.Conclusions MRS can manifest the regional cerebral metabolic changes of rabbits with explosive injury at different time points and hence provide a theoretical basis for understanding the local tissue changes and determining the type of tissue damage after blast injury.

Objective To investigate the effects of biatrial infusion on pulmonary artery pressure (PAP)after cardiopulmonary bypass (CPB) in patients undergoing mitral valve replacement.Methods Twenty NYHA Ⅱ or Ⅲ patients aged 22-53 yr weighing 34-57 kg undergoing mitral valve replacement complicated by pulmonary hypertension (mean pulmonary artery pressure (MPAP) ＞ 50 mm Hg) were randomly divided into 2 groups ( n = 10 each): infusion via right atrium group (group R) and infusion via both atria group (group B). After induction of anesthesia, a three cavity floating Swan-Ganz catheter was placed via right internal jugular vein to monitor CVP,PAP, pulmonary capillary wedge pressure (PCWP) and CO. The patients received infusion of prostaglandin E1 30-150 ng· kg- 1 · min - 1 and phenylephrine 0.2-0.6 μg· kg- 1 · min- 1 via central veins in group R and infusion of prostaglandin E1 30-150 ng·kg-1 ·min-1 via central veins and phenylephrine 0.2-0.6 μg·kg-1 ·min-1 via left atrium in group B. MAP, HR, MPAP, PCWP, CVP and CO were recorded 5 min before administration (T0), and 5,10, 30 and 60 min after administration (T1-4). Pulmonary vascular resistance index (PVRI), systemic vascular resistance index (SVRI) and CI were also calculated. Results D:\1111111111\MDB\zhmzxzz98201008.mdbCompared with the value at T0, MAP, MPAP, PCWP and PVRI were significantly decreased, while CI was increased at T1-4 in group R, and MAP, CI and SVRI were significantly increased, while HR, MPAP, PCWP, CVP and PVRI decreased at T1-4 in group B ( P ＜ 0.05).MAP, CI and SVRI were significantly higher, while HR, MPAP, PCWP, PVRI and CVP lower in group B than in group R ( P ＜ 0.05). Conclusion Biatrial infusion can effectively reduce PAP and pulmonary vascular resistance after CPB in patients undergoing mitral valve replacement.

Objective To study the characteristics of the sciatic nerve injuries inflicted by highvelocity triangle fragments in swine.Methods According to the injury position,14 swines weighing (34.29±5.2)kg were divided into two groups.All swines were shot by 0.37 g triangle fragments at (773.1±12.4)m/s aimed at the midpoint of the lateral body surface projection of the right sciatic nerve in Group A(n=7)and at 2 cm aside from sciatic nerve in Group B(n=7).The entrance,exit and length of wound tract,distance between nerve and wound tract were detected,and the pathology of sciatic nerve and wound tract at 48 hours after injury were observed under light microscope and electron microscope.Results The course of the fragments was deviated to different extent.Six sciatic nerves were in primary wound tract,four of which were lacerated and ruptured at different degrees.The light microscope showed pathological changes including severe hemorrhage and edema,leukocytic infiltration,neurotmesis,axonotmesis and light myelin coloration;while electron microscope showed severe degeneration of myelin sheath and neuraxon distortion.Six nerves(one in Group A and five in Group B)were in the concussion zone,with the distance between sciatic nerve and primary wound tract for(2.07±0.45)cm.Hyporrhea and hyperemia under perilemma and perineurium,partial nerve fiber disrupt and axonotmesis were observed by light microscope and partly myelin sheath delamination by electron microscope.Two nerves (one in Group A and another in Group B)were in the contusion zone,with moderate pathological changes.Conclusions The track of high velocity triangle fragments is unstable in the tissues.High disrupt rate,severe and wide extent lesion,early Wallerian degeneration are the main characteristics of direct injuries of swine's sciatic nerves.Indirect injuries can induce pathologic changes too,for which the lesion severity is related to the distance between nerves and primary wound tract.