10.3969/j.issn.2095-4344.2013.26.002

BACKGROUND:Kienb?ck disease lacks of suitable animal models, which are similar to the pathological process of avascular necrosis of human lunate bone.
OBJECTIVE:To establish a new animal model of Kienb?ck disease using medical TH adhesive embolism and to explore the rationality of model establishment.
METHODS:A total of 30 healthy adult New Zealand rabbits, male or female, were selected. Using self-control method, the rabbits were randomly assigned to experimental sides and control sides. By dril ing in the center of the lunate bone, 0.2 mL of medical TH glue was injected three times. An equal volume of physiological saline was injected into the center of the lunate bone on the control side. X-ray examination, general observation, Micro-CT measurement of bone, and tissue pathology detection were conducted at 4, 8 and 12 weeks.
RESULTS AND CONCLUSION:Gross specimen, X-ray and histological results showed that ischemic necrosis of the lunate bone on the experimental side was visible at 8 weeks after model induction. The ischemic necrosis of the lunate bone became more typical at 12 weeks. Among the Micro-CT microscopic parameters of trabecular bone, trabecular bone density parameters bone volume fraction and the number of trabecular bone were significantly lower on the experimental side than those on the control side (P<0.05). Spatial parameters of trabecular bone significantly increased. Trabecular separation and structure model index on the experimental side were significantly greater than those on the control side. Results suggested that ischemic necrosis of the lunate bone appeared on the experimental side at 8 weeks after injection of medical TH glue. Rabbit models of ischemic necrosis of the lunate bone can be established at 12 weeks. Thus, alterations, which were similar to ischemic necrosis of human lunate bone, appeared, such as blood transportation damage in the lunate bone, trabecular bone fracture, and empty lacuna, when surrounding tissues were not obviously injured.

Female ; Humans ; Infant ; Meningitis, Meningococcal ; diagnosis

Objective To explore the clinical,radiological findings and pathogenic factor of spondylometaphyseal dysplasia.Methods Five cases were reported and the relevant documents were studied retrospectively.Plain X-ray film was performed in all patients.Results There were 4 male and 1 female,age ranged from 4 to 15 years with average of 9 years.The main features of 5 cases included delayed bone age,stature short,short trunk,waddling gait noted,scoliosis in 2 cases,kyphoscoliosis in 1 case,severe genu valgum in 2 cases.The main X-ray appearance of 5 cases is multiple irregularities of long bones metaphyses associated with platyspondylia,epiphysis is normal.Type Ⅰ SMD in 2 cases,Type Ⅱ SMD in 1 case,Type Ⅲ SMD in 2 cases.Conclusion When we meet children with delayed bone age and stature short,and muhiple irregularities of long bones metaphyses associated with platyspondylia were seen in X-ray plain film.we should think about spondylometaphyseal dysplasia.

Abstract: To analyze the clinical, therapeutic and laboratory characteristics of disseminated cryptococcosis caused by Cryptococcus neoformans invading the blood stream in patient with liver cirrhosis and splenectomy. A 30-year-old male underwent splenectomy plus pericardial devascularization due to "splenomegaly and hypersplenism" in March in 2016. The patient had intermittent fever after operation for many times, and successively accompanied with back pain, left lower limb abscess and right hip pain. The highest body temperature was 39 ℃. CT and MRI revealed the lung lesion and multiple bone destruction. During that period, the effect of antibiotics was not good. On April 19th, 2017, Gram's stain, India ink stain, API 32C, Vitek 2 Compact, ribosomal ITS and IGS sequence analysis were performed to identify the strain isolated from the pus and blood stream. The serum of the patient was detected for cryptococcal antigen. Antifungal susceptibility test was used to determine drug sensitivity and minimum inhibitory concentration (MIC). The Cryptococcus neoformans isolated from fresh pus specimen showed a prominent, thick capsule after India ink stain. The colonies isolated from pus and blood stream were identified Cryptococcus neoformans using API 32C, Vitek 2 Compact, and sequence analysis of rDNA ITS and IGS. Cryptococcal capsule antigen was positive. The minimal inhibitory concentrations of 5-Flucytosine, amphotericin B, fluconazole, itriconazole, voriconazole against the isolate were <4 μg/mL, <0.5 μg/mL, 4 μg/mL, ≤0.25 μg/mL, 0.125 μg/mL respectively. The patient was initially treated with intravenous amphotericin B and flucytosine. After anti-Cryptococcus treatment for two months, the patient clinically improved, and the lesions were reduced on a follow-up CT scan. The patient made a full functional recovery after treatment for six months. Cryptococcosis has hidden onset, atypical clinical symptoms and lack of specificity. Blood stream is the main channel for Cryptococcus to spread and involve many organs of the whole body, including skin, bone and so on. Therefore, early use of blood culture to monitor blood flow dissemination, actively removing the primary focus and cutting off the infection route in time and carrying out effective anti-Cryptococcus treatment are conducive to the patient's early recovery.

BACKGROUND: Wharton's Jelly-derived mesenchymal stem cells are relatively primitive stem cells that are ideal vectors for gene therapy. However, there is a lack of studies on the conditions for the electrotransfection of Wharton's Jelly-derived mesenchymal stem cells. Therefore, exploring the optimal conditions for the electrotransfection of Wharton's Jelly-derived mesenchymal stem cells occupies an important position. OBJECTIVE: To investigate the effects of different electroporation conditions on the transfection efficiency of Wharton's Jelly-derived mesenchymal stem cells, and to explore the optimal conditions for cell electroporation. METHODS: By controlling the transfection conditions such as voltage, pulse duration and cell status, EEV-EGFP plasmids were transfected into Wharton's Jelly-derived mesenchymal stem cells by electroporation under different conditions. Transfection efficiency was detected by flow cytometry. RESULTS AND CONCLUSION: (1) The transfection efficiency was intended to increase when the voltage ranged from 125 V to 150 V, and the maximum transfection efficiency was obtained when the voltage was 150 V. However, when the voltage was further increased to 170 V, the transfection efficiency began to decrease considerably. (2) The maximum transfection efficiency was obtained when the pulse duration was 5.0 ms, while it was certainly decreased when the pulse duration was 2.5 and 7.5 ms. (3) The transfection efficiency of the cells cultured under normoxia was higher than that under hypoxic culture. These findings reveal that normally cultured Wharton's Jelly-derived mesenchymal stem cells can achieve higher electroporation efficiency via two pulse sessions at a voltage of 150 V, pulse duration of 5.0 ms, and pulse interval time of 50 ms.

BACKGROUND: Mesenchymal stem cells are a kind of adult stem cells with self-renewal and multi-directional differentiation potential. These cells have the functions of immunoregulation, regulation of cell growth and repair of injury. In recent years, it has been found that when inflammatory injury occurs, mesenchymal stem cells can regulate the secretion of inflammatory factors, and function on the damaged region, thereby repairing and improving tissue damage caused by inflammation. OBJECTIVE: To review the immunoregulatory effects of mesenchymal stem cells on T lymphocytes, B lymphocytes, dendritic cells and natural killer cells when the body is in an inflammatory state. METHODS: A computer-based search of PubMed and CNKI was performed for articles published from 2000 to 2018 using the keywords of"inflammatory microenvironment, mesenchymal stem cell, immune response, T cell, B cell, DC, NK cell" in English and Chinese, respectively. Seventy-six articles related to the topic and with reliable arguments were finally included in result analysis. RESULTS AND CONCLUSION: The interaction between mesenchymal stem cells and inflammatory cells determines the result of tissue damage repair. In an inflammatory state, the biological characteristics of mesenchymal stem cells will undergo certain changes, but mesenchymal stem cells can still exert immunomodulatory effects by secretion of various soluble cytokines or via cell contact. There are still many problems to be further explored to facilitate better clinical application of mesenchymal stem cells.

Objective To investigate the value of PET/CT imaging of cerebral glucose metabolism (CGM)and cerebral blood flow (CBF)in evaluating chronic disorders of consciousness (CDC).Methods A total of 10 CDC patients (5 males,5 females,age (50.9 ±17.2)years)and 10 healthy controls (5 males,5 females,age (52.0±10.3)years)from January 2016 to March 2017 were recruited to perform brain PET/CT of CGMand CBF.The brain PET imaging using 13 N-Ammonia was performed and followed by 18 F-fluorodeoxyglucose (FDG)PET.The mean standardized uptake values (SUVmean )of frontal,parietal, temporal and occipital lobes as well as basal ganglia,thalamus were obtained.The SUVmean of cerebral re-gions/SUVmean of cerebellum ratios (SUVr )were acquired.The SUVr were compared between the patients and controls.The imaging characteristics of CGM and CBF were investigated,and their relationships with clinical scores were further analyzed.Two-sample t test and Pearson correlation analysis were used to analyze the data.Results The radioactive distribution in the brain of healthy controls was symmetrical.SUVr of cer-ebral regions in the affected side of patients were significantly lower than those of the controls both in CGM imaging and CBF imaging (t values:2.90-5.19,all P<0.05).In 10 CDC patients,there were 9 with hypo-metabolism in basal ganglia and thalamus,8 with hypometabolism in frontal and parietal lobes,and 7 with hypometabolism in temporal and occipital lobes.At the same time,there were 7 with parietal hypoperfusion and 6 with hypoperfusion in other cerebral regions in the CDC patients.In the frontal,parietal lobes and basal ganglia,the CGMand CBF were both correlated with the clinical scores (r values:0.473-0.606,all P<0.05).Abnormal metabolism-perfusion patterns were divided into 3 types.Type Ⅰ included 2 patients and their hypometabolism and hypoperfusion were mismatched completely.Type Ⅱ included 3 patients and their hypometabolism and hypoperfusion were matched in frontal,parietal,occipital and temporal lobes,while mismatched in basal ganglia and thalamus.Type Ⅲ included 5 patients and their hypometabolism and hypoperfusion were matched completely.The clinical scores of typeⅠ,Ⅱand Ⅲ were 10.5,8.3 and 5.6, respectively.Conclusion The PET/CT imaging of cerebral blood flow and metabolism is useful in evalua-ting the disorders of consciousness.

Lysine decarboxylase is a key enzyme involved in the upstream biosynthesis of lycopodium alkaloids (LAs) such as huperzine A, contributing to the decarboxylation of lysine to 1,5-pentanediamine (cadaverine). Three lysine decarboxylase genes (HsLDC-L1, HsLDC-L2, HsLDC-L3) were successfully cloned from Huperzia serrata using transcriptomic sequence data mining strategy combined with reverse transcription PCR. The physicochemical properties, secondary and tertiary structures, amino acid identities, and evolutionary relationship of the three LDCs were analyzed by online bioinformatics analysis platforms and DNAMAN, MEGA 7.0 software, revealing that all of these proteins had the conserved PLP binding domain and active site residues were completely conserved in LDCs. Phylogenetic analysis showed that these LDCs were located in the same branch as other known LDCs from LA-producing plants. Accordingly, the ORFs of these three HsLDCs were inserted into different expression plasmids for further expression in E. coli. However, only HsLDC-L1 was successfully expressed in E. coli BL21 (DE3) by inserting into a pCold TF vector. The recombinant protein was purified by Ni²⁺ affinity chromatography purification. HsLDC-L1 contains 469 amino acid residues, with a calculated molecular weight of 50.50 kDa. HsLDC-L1 expectedly catalyzed the decarboxylation of lysine to produce cadaverine. In addition, HsLDC-L1 can also catalyze the generation of putrescine from ornithine. However, it cannot catalyze the decarboxylation of tyrosine, phenylalanine, tryptophan and histidine. The results not only provide insight into the biosynthesis of LAs including huperzine A, but also provide a critical genetic element for the overproduction of Δ¹-piperideine and pelletierine, the essential biosynthetic precursors of LAs, using synthetic biology strategies.