BACKGROUND: There are a lot of immunologic studies about limb allotransplantation in animal experiment. But, it is only early investigation in clinic; its clinical immunologic study needs further accumulation.OBJECTIVE: To dynamically analyze the early immunologic state change in patients following hand allotransplantation.DESIGN: Controlled trial.SETTING: Department of Orthopaedics, Guangzhou General Hospital,Guangzhou Military Area Command of Chinese PLA; Department of Traumatic Orthopaedics and Department of Laboratory Medicine, Nanfang Hospital, First Military Medical University of Chinese PLA.PARTICIPANTS: Two patients who underwent unilateral hand allotransplantation in the Department of Orthopaedics, Guangzhou General Hospital,Guangzhou Military Area Command of Chinese PLA were enrolled, serving as experimental group. The observation was between September 1999 and March 2000. Twenty persons, including 12 male and 8 female, who homochronously received health examination, aged 20 to 45 years, were enrolled, serving as healthy control group. They all had no reactive immune and infectious diseases, and voluntarily participated in the trial.METHODS: Peripheral blood was collected from 2 patients who underwent hand allotransplantation once respectively at pre-operative 1 day and 3 days. Blood collecting was performed once per day at post-operative 1 week, three times per week at post-operative 2 to 4 weeks, twice per week at 5 to 8 weeks post-operation, once per week at 9 to 16 weeks post-operation, twice per month at 5 to 6 months post-operation. ① Peripheral blood T cell subgroups (CD3+,CD4+,CD8+T cells)were detected by flow cytometer,serum panel reactive antibody (PRA) by ELISA method, serum C-reactive protein by turbidimetric immanoassay (TIA), serum creatine kinase (CK) by enzyme dynamics method. ②Mixed lymphocyte reaction(MLR): mitomycin C-treated donor peripheral blood lymphocytes were used as stimulator, and proliferative reaction of peripheral blood lymphocyte of patients to donor transplanted antigen was detected with the incorporation of 3H-TDR method (Negative: There was no significant difference between the mean value of stimulation index and 1, conversely positive). Autogenic peripheral blood lymphocytes treated with the same way replaced donor stimulator, serving as control. Stimulation index of each specimen was calculated (Stimulation index=Experiment cmp/controlcpm), serving as control index. Peripheral blood T-cell subgroups (CD3+,CD4+,CD8+T cell), serum PRA, C-reactive protein and CK were detected in 20 persons in healthy control group;Twenty persons were randomly divided into 10 groups. Two persons in each group were used as donor and recipient mutually and performed MLR.MAIN OUTCOME MEASURES: ① Peripheral blood T cell subgrpups (CD3+,CD4+,CD8+T cell). ②PRA. ③ C-reactive protein. ④CK. ⑤MLR.RESULTS: ①CD3+,CD4+,CD8+T cell levels were obviously decreased within one week after operation. CD3+ and CD4+T cell levels both recovered to be the pre-operative levels, but CD8+ level exceeded pre-operative level significantly [CD3+: (66.43±4.56); CD4+: (30.55±3.94); CD8 +:(33.45 ±2.69)]. There was no significant difference between experiment group and control group. ②Serum PRA was 0 to 10%, there was no significant difference as compared with control group. ③ Serum C-reactive protein was 0 to 0.359 mg/L, there was no significant difference as compared with control group. ④ Serum CK was 25 to 170 mmol/L, and there was no significant difference as compared with control group. ⑤ MLR after transplantation was negative, and it turned into be positive 5 months later.They were all positive in control group.CONCLUSION: Short-term change and long-term redistribution of T cell subgroups are closely related to immunosuppressive agent, suggesting that immunosuppressive agent has obvious effect on T-cell subgroup following hand allotransplantation. Immuno-induction schedule make patients be in immune suppression state, which effectively avoid early rejection. But patients cannot bear specificity yet; they need the inhibition of immunosuppressive agents.

Abstract: Objective To analysis the genetic evolution characteristics of hemagglutinin (HA) gene of influenza A(H1N1)pdm virus in Changsha City from 2016-2023, to understand the trend of the HA genetic evolution and the mutations of the amino acid. It provides a scientific basis for the prevention and control of influenza epidemics, as well as the screening of vaccines under the new situation. Methods The A(H1N1)pdm09 virus strains from Changsha City from 2016 to 2023 were isolated using SPF chicken embryos, and then the HA genes were sequenced by MiSeq of Illumina Inc. The homology of HA gene was analyzed by MegAlign of the DNASTAR, and the phylogenetic tree was constructed using the Neighbor Joining (NJ) method in the Molecular Evolutionary Genetics Analysis version 11 (MEGA11). Results The homology of the HA gene of A(H1N1)pdm virus in Changsha from 2016 to 2023 was between 94.8%-99.9%, with the HA gene homology decreasing annually. The homology between the isolated strains of A(H1N1)pdm09 in Changsha City from 2016 to 2023 and the WHO recommended vaccine strain ranged from 96.8% to 99.0%, indicating a relatively good match between the flu isolates and the recommended vaccine strain. The phylogenetic tree of the HA gene of the A(H1N1)pdm09 influenza virus in Changsha City showed that the HA gene evolved into several different branches within the 6B branch, and it had currently evolved to 6B.1A.5a.2a branch. Constant mutations had occurred at the amino acid sites of the four antigenic determinant clusters of HA protein. Currently, amino acid mutations had occurred at 15 antigenic sites within the four antigenic determinant clusters, and the newly emerged A186T antigen mutant site in the isolates from 2023 was worth recent notice. The receptor-binding sites are relatively conserved in loop 130, minor amino acid mutations occurred in loop 220, whether the amino acid mutation site in loop 190 is becoming more stable needs to be further monitored. Taking A/California/07/2009 (CY121680) as the reference strain, most of the A(H1N1) pdm09 isolates in Changsha was increased 162 NQTY glycosylation site and was decreased 276 NTTC glycosylation site, and the glycosylation mutations at these two sites have become more stable recently. Conclusions The HA genes of influenza A(H1N1)pdm virus in Changsha are constantly evolving and mutating, suggesting influenza surveillance should be strengthened continuously.

Objective To explore the impact factors for post-thaw embryo survival rate and clinical pregnancy rate in frozen-thawed embryo transfer program. Methods The clinical data of 573 cycles of frozen-thawed embryo transfers were retrospectively analysed. Groups were divided according to the pre-freeze embryo quality, pre-freeze embryonic developmental stage, frozen-thawed embryo quality and cryopreservation technique, respectively, and post-thaw embryo survival rates and/or clinical pregnancy rates were compared among groups. Results The clinical pregnancy rate of high quality pre-freeze embryo was significantly higher than that of low quality pre-freeze embryo (31.8% vs 20.0%) (P< 0.05). There was no significant difference in the post-thaw survival rates and clinical pregnancy rates between embryos frozen at day 2 of ferrtilization and those frozen at day 3 of ferrtilization(79. 1% vs 82.9% and 25.5% vs 31.2%, respectively) (P>0.05). The clinical pregnancy rates of the transfer cycles only with fully intact embryos and with mixed embryos were significantly higher than that only with partially damaged embryos(36.7% vs 24.1% and 29.2% vs 24.1%, respectively)(P<0.05). The post-thaw survival rate and post-thaw high-quality embryo rate were significantly higher in those processed with modified cryopreservation technique than in those processed with original cryopreservation technique (82.0% vs 66.3% and 50.0% vs 27.5%, respectively)(P<0.05). Conclusion Pre-freeze embryo quality, post-thaw embryo survival rate and post-thaw embryo quality have a positive correlation to subsequent clinical pregnancy rate. Favorable cryopreservation technique may ensure the success of post-thaw embryo recovery and transfer.

Objective To investigate the choroidal thickness (CT) in patient (＞ 50 years old) with central serous chorioretinopathy (CSC) compared to age-matched healthy controls.Methods Together 25 (＞ 50 years old) patients with CSC from January 2016 to January 2017 were enrolled in these study.Of 25 patients,20 patients were affected unilaterally and 5 patients were affected bilaterally.These patients were divided into affected eye group (30 eyes) and unaffected fellow eye group (19 eyes).Addition 26 age-matched healthy subjects were recruited as control group (26 eyes).Enhanced depth imaging technique of optical coherence tomography (EDI-OCT) was used to measure the choriodial thickness at 5 sites,including subfovea and 500 μm,1500 μm temporal and nasal to the fovea.Results The mean age of patients with CSC was (60.08 ± 7.68) years.The subfoveal choroid thickness of both affected eye group and unaffected fellow eye group was significantly thicker than that of the control group [(414.17 ±85.88)pan vs.(352.89 ±97.22) μn vs.(280.69 ±90.99) pau] (both P ＜0.05).In the patients with monocular affected eyes,the subfoveal choroid thickness of the affected eyes was thicker than that of the fellow eyes [(414.07 ± 85.88) μm vs.(352.89 ± 97.22) μm] (P ＜ 0.05).There were significant differences in choriodal thickness at other sites in the three groups (all P ＜ O.05).Conclusion The choriodal thickness in patients with CSC at over 50 years of age is significantly thicker than that in the agematched healthy subjects,with subfoveal choroid thickness ＞ 400 μm,which is helpful for the differential diagnosis of elderly patients with macular serous retinal detachment.

Objective To explore the effect of L-carnosine on neuronal cell apoptosis in young rats with experimental febrile seizures(FS).Methods Forty 15-day SD rats were randomly divided into intervention group(n=30)and FS group(n=10).Warm water was used to induce 10 times FS.The intervention group was divided into E,G and H group,10 rats in each group.Intraperitoneal injection of L-carnosine(250 mg/kg)was separately given to the rats in E group,G group and H group respectively after 30,60 and 120 min of seizure.FS group were induced FS,but they were not given intervention.The rats were sacrificed at 12 hours after the last seizure.Neuronal cell apoptosis was determined by terminal eoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)in situ cell death kit.TUNEL positive cells were stained and counted as apoptosis in hippocampus and cortex.Ultrastructural changes of apoptosis neurons were observed under the electron microscope.Results The neuronal cells apoptosis count was 25.37?1.95 in FS group,12.36?1.13 in E group,17.85?2.04 in G group,and 22.69?2.69 in H group.Neuronal apoptosis of FS group was apparently higher than that of interventional groups(F=10.75 P0.05).Under the electron microscope,neuronal damage on hippocampal CA1 area and dentate gyrus of FS group and H group was obviously higher than that of E group.Conclusions Early injection of L-carnosine would not only relieve neuronal apoptosis of repeated FS,but also play a role in the protection of neuronal cells.

0.05).Conclusions Early injection of L-carnosine would not only improve cerebral oxidative phosphorylation,relieve neuronal injury of repeated FS,but play a role in the protection of neuronal cells.

OBJECTIVETo discuss the clinical effects of open reduction and internal fixation (ORIF) for treatment of patients with Lisfranc injury combined the second metatarsal base comminuted fracture.

METHODSFrom March 2007 to June 2012, 7 patients with Lisfranc injury combined the second metatarsal base comminuted fracture were treated including 5 males and 2 female aged from 22 to 51 years old (means 42 years), 4 of sprain and 3 of traffic injury. According Myerson classification, there was 1 case of type A, 3 of type B and 3 of type C. Kirschner wire was used to fix Lisfranc ligament placing from the medial cuneiform bone to the second metatarsal base during the operation. After the operation American Orthopaedic Foot and Ankle Society (AOFAS) criteria system were applied to evaluate the foot and ankle function. Preoperative and postoperative AP, lateral and oblique X-ray and CT scan were collected for radiographic evaluation.

RESULTSAll patients were followed up from 12 to 20 months (16.8 months in average). According to AOFAS criteria system, 3 cases were excellent result,3 good, 1 fair. All the wounds were primary healing without skin necrosis, infection, Kirschner loose,broken, or other complications.

CONCLUSIONKirschner wire had good clinical efficacy for fixing Lisfranc ligament injury with the second metatarsal base comminuted fracture, and could avoid arthrodesis.

Adult ; Bone Wires ; Female ; Humans ; Male ; Metatarsal Bones ; injuries ; surgery ; Middle Aged ; Tarsal Joints ; injuries ; surgery ; Wound Healing

Objective To judge method performance of routine biochemistry parameters on Roche Modular PPI testing system by using westgard's method evaluation decision chart.Methods We assessed imprecision(CV)from internal quality control and inaccuracy(bias)from external quality control evaluation.Combined estimates of imprecision and inaccuracy by plotting imprecision as the x-coordinate and inaccuracy as the y-coordinate to locate an expected operating point of every item on the chart.By comparing this operating point with allowable total errors(TEa),we can decide whether the performance is acceptable or not.Results In the 27 different parameters tested,imprecision and bias of calcium were 0.08 mmol/L and 0.06 mmol/L respectively,its performance was marginal.The imprecision of creatinine,urea,glucose, sodium,chloride and phosphorus were 3.20%,2.13%,1.52%,0.89 mmol/L,1.10% and 1.55%,the bias were 4.79%,0.96%,4.63%,0.80 mmol/L,1.74% and 4.13% respectively,their performance was good.M1 other 20 items were of excellence performance.Conclusions Routine biochemistry parameters on Roche Modular PPI testing system possessed good precision and accuracy,and their performance were acceptable.To judge method performance of biochemistry testing system by using westgard' s method evaluation decision chart was easy to do and suited for clinical laboratory.

Objective In order to explore the prevalence of chronic obstructive pulmonary disease (COPD)among residents aged over 40 in Yongjia County,and to provide basic data for the prevention and control strategies and measures of chronic obstructive pulmonary disease.Methods Residents were investigated by questionnaire investigation,body measurements, and pulmonary function tests.Questionnaire survey was including demographic information,chronic obstructive pulmonary disease knowledge awareness,personal and family history of the disease,respiratory symptoms,case management of respiratory diseases,risk factors.Body measurements were including height,body weight,waist circumference,blood pressure and heart rate.Pulmonary function tests was including one second forced expiratory volume (FEV1 ),forced expiratory volume (FEV6 ) and forced vital capacity (FVC ).Results A total of 585 residents were investigated, including 85 patients with chronic obstructive pulmonary disease,and the prevalence rate of 1 4. 53%.Male chronic obstructive pulmonary morbidity was higher than female (χ2 =44. 29,P=0. 001 ),and with age increased,the prevalence rate increased (χ2 =1 9. 56,P<0. 001 ).Single factor analysis showed that the main risk factors were male,age ≥40, often cough /expectoration,smoking,occupational exposure history.Multivariate logistic regression analysis showed that male (OR=1 . 962,95%CI:1 . 025 -3. 757),expectoration (OR=2. 346,95%CI:1 . 1 48 -4. 794)and age (in the age group of 50:OR=2. 561 ,95%CI:1 . 221 -5. 372;age≥60(OR=7. 438,95%CI:3. 601 -1 5. 361 )were the risk factors of chronic obstructive pulmonary disease (COPD).Conclusion Chronic obstructive pneumonia has become a public health problem that affects the health of the residents.We should take effective preventive measures against the risk factors.

OBJECTIVETo investigate the effects of oxidized low-density lipoprotein receptor 1 (LOX-1) on secretion of adhesive molecules mediated by ox-LDL in human umbilical endothelial cells (HUVECs).

METHODSHUVECs with different concentration of ox-LDL (0, 10, 20, 50, 100 microg/ml) were incubated for 24 h, or HUVECs were pretreated with 250 microg/ml poly (I) or 250 microg/ml carrageenan for 2 h and then incubated with 50 microg/ml ox-LDL for another 24 h. Expression of LOX-1 was determined by realtime RT-PCR and Western blot. mRNA and protein of ICAM-1, VCAM-1 and E-selectin were examined by RT-PCR and Western blot respectively.

RESULTSIncubation of HUVECs with ox-LDL (10-100 microg/ml) enhanced the expressions of LOX-1, ICAM-1 and E-selectin in a concentration-dependent manner (P < 0.01). On the contrary, ox-LDL did not affect the expression of VCAM-1 by HUVECs. The expression of LOX-1, ICAM-1 and E-selectin induced by ox-LDL were reduced in HUVECs pretreated with 250 microg/ml poly (I) or 250 microg/ml carrageenan for 2 h and then incubated with 50 microg/ml ox-LDL for 24 h. This showed that both poly (I) and carrageenan obviously decreased the expression of LOX-1, ICAM-1 and E-selectin induced by ox-LDL.

CONCLUSIONox-LDL may upregulate the expression of LOX-1, ICAM-1 and E-selectin, and LOX-1 blocker may partly inhibit this upregulation. The results suggest that the expression of inflammatory molecules induced by ox-LDL in HUVECs is mediated by LOX-1.

Cell Adhesion ; Cell Adhesion Molecules ; Cells, Cultured ; E-Selectin ; metabolism ; Endothelial Cells ; metabolism ; Endothelium, Vascular ; metabolism ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; Lipoproteins, LDL ; biosynthesis ; RNA, Messenger ; metabolism ; Receptors, Oxidized LDL ; metabolism ; Scavenger Receptors, Class E ; metabolism ; Umbilical Veins ; cytology ; Vascular Cell Adhesion Molecule-1 ; metabolism