BACKGROUND:Microencapsulated cels are commonly used as a tool to overcome immune rejection after subarachnoid transplantation. However, the effect of microencapsulation on the secretion of human pheochromocytoma cels is unclear.
OBJECTIVE:To observe the growth and secretion of primarily microencapsulated cultured human pheochromocytoma cels in artificial cerebrospinal fluid.
METHODS: The human pheochromocytoma tissues were digested successively to isolate human pheochromocytoma cels that were then cultured in artificial cerebrospinal fluid. Primary cels were covered with alginate-polylysine-alginate microcapsules, and then the cel morphology was observed with inverted phase contrast microscope. Levels of met-enkephalin and norepinephrine in cel culture medium were detected by enzyme-labeled immunosorbent assay (ELISA). We used cel counting kit-8 colorimetric assay to obtain the growth curve of human pheochromocytoma cels in artificial cerebrospinal fluid.
RESULTS AND CONCLUSION:Microcapsulated human pheochromocytoma cels were in suspension and the process outgrowth increased slowly. Compared with non-microcapsulated cels, the proliferation rate of microcapsulated cels increased significantly. ELISA results revealed a significant increase in the levels of met-enkephalin and norepinephrine secreted from the microencapsulated cels compared to the non-microcapsule group. There was a wide variation in contents of met-enkephalin and norepinephrine from different tumors. These findings indicate that microencapsulated human pheochromocytoma cels can survive wel and have good secretion function in artificial cerebrospinal fluid, and human pheochromocytoma cels from different tumor tissues have stable secretory function.

BACKGROUND:Lumbar spine MRI and electrophysiological test are reliable methods for evaluating nerve root injury caused by lumbar disc herniation.
OBJECTIVE:To analyze the correlation between the MRI-based grading system and the latency and frequency of F wave as wel as latency and amplitude of H-reflex in patients with lumbar disc herniation.
METHODS:MRI imaging of the lumbar spine was performed with a 3.0-T imager and a dedicated TCL coil to classify lumbar disc herniation and nerve root compression. F wave and H reflex were detected on the patient bilateral tibial nerves using Oxford myoelectricity evoked potential instrument.
RESULTS AND CONCLUSION:Spearman correlation analysis showed that the MRI-based grading of patients with lumbar disc herniation had a negative correlation with F wave frequency (r=-0.594 0, P<0.000 1), and a positive correlation with F wave latency (r=0.825 6, P<0.000 1) and H-reflex latency (r=0.875 0, P<0.000 1), but no correlation with H-reflex amplitude (R=0.117 4, P=0.257 3). With MRI grading increased, F wave frequency was decreased, and F wave and H-reflex latency were prolonged gradual y, indicating aggravating nerve root compression.

BACKGROUND: Exercise has been proved to accelerate the proliferation of intervertebral disc cells and extracellular matrix production in healthy rats. For the degenerative intervertebral disc, whether exercise also has positive effects on its cell proliferation, extracellular matrix production or pain relief remains unclear. OBJECTIVE: To investigate the effect of exercise on the extracellular matrix production in a rat model of intervertebral disc degeneration.METHODS: A rat model of intervertebral disc degeneration was prepared by Freund's complete adjuvant injection into the intervertebral disc at L5-6 levels. Then, the model rats were allowed to have a rest for 2 weeks. All rats were then randomly divided into exercise and control groups. Rats in the exercise group were forced to run every day, while the controls allowed free activities in the cage. The behavioral tests were performed at 7, 14, 28, 42, 56 and 70 days after modeling; meanwhile, the intervertebral disc samples were collected used for alcian blue staining and immunohistochemical staining to detect the levels of proteoglycan, aggrecan and collagen type Ⅱ in the intervertebral disc cells, respectively.RESULTS AND CONCLUSION: Vocalization threshold on the rat back of punctured disc was significantly decreased, while grooming and wet-dog shaking were significantly increased at 7 days after modeling compared with the baseline (P < 0.05), suggesting that Freund's complete adjuvant injection successfully induces disc degeneration, hyperalgesia and abnormal behaviors. Further, the vocalization threshold and wet-dog shaking in the exercise group showed significant improvement compared with the control group after 14 days of exercise (P < 0.05), while the grooming was significantly reduced until the 28th day (P < 0.01), indicating that exercise can alleviate pain caused by disc degeneration in model rats. At 21 days after modeling, the levels of proteoglycan, aggrecan and collagen type Ⅱ in the nucleus pulposus and annulus fibrosus were significantly decreased compared with the baseline (P < 0.01), indicating the occurrence of disc degeneration. After 14 days of training, the levels of proteoglycan, aggrecan, and collagen type Ⅱ in the nucleus pulposus and annulus fibrosus in the exercise group were significantly increased compared with the control group (P < 0.01). Moreover, after 8-week exercise, the level of proteoglycan in the nucleus pulposus and annulus fibrosus in the exercise group was increased by 4-5 times compared with the control group, and levels of aggrecan and collagen type Ⅱ in the nucleus pulposus in the exercise group also was increased by 3-4 times compared with the control group. To conclude, exercise can promote extracellular matrix increased by production by increasing the levels of proteoglycan, aggrecan, and collagen type II in the degenerative intervertebral disc.

OBJECTIVE: This study was designed to explore the risk factors associated with severity of juvenile onset recurrent respiratory papillomatosis. METHOD: A retrospective study was conducted to study determinants of severe forms of juvenile recurrent onset respiratory papillomatosis. The patients were separated into different groups based on the onset age, the first recurrence of age, the first recurrence of period, gender and incision of tracheal respectively. The relationship among the lesion severity score,the involvement of the subregion, operation period and the next operation period were also explored. RESULT: It was observed that some children who recurred before 4 years old required more surgery, shorter operation period(the average, longest or shortest operation period) than those elder children, the differences were statistically (P=0. 029, 0. 003, 0. 010, 0. 039, respectively). The severity score of lesion was correlated positively with the involvement of the subregion and negatively with operation period (r=0. 914, -0. 451, respectively). Some children who diagnosed before 4 years old had to endure more severity score and shorter operation period than those older children, the differences were statistically (P= 0. 036, 0. 000, respectively). 8 cases accepted incision of tracheal, they accepted more surgery too. But the differences in the onset age, the first recurrence of age, and the operation period were not statistically. CONCLUSION The results showed that the clinical course of juvenile onset recurrent respiratory papillomatosis was closely related to the first recurrence age and period, while the severity of disease was associated to the onset age and the involvement of the subregion.
Adolescent ; Age of Onset ; Child ; Child, Preschool ; Humans ; Papilloma ; Papillomavirus Infections ; classification ; epidemiology ; surgery ; Respiratory Tract Infections ; classification ; epidemiology ; surgery ; Retrospective Studies ; Risk Factors ; Severity of Illness Index ; Trachea

A reversed-phase HPLC method for simultaneous determination of gatrorrhizine, columbamine, epiberberine, coptisine, palmatine and berberine in Coptis chinensis was developed. Analysis was carried out on an Xtimate C18 column (4.6 mm x 250 mm, 5 microm) eluted with acetonitrile-30 mmol x L(-1) ammonium bicarbonate solution (including 0.7% ammonia and 0.1% triethylamine) by gradient elution. The detective wavelength was 270 nm, the column temperature was 30 degrees C, and the flow rate was 1.0 mL x min(-1). By the above method, the linear ranges of gatrorrhizine, columbamine, epiberberine, coptisine, palmatine and berberine were 0.85-16.96 (r = 0.9997), 1.25-24.96 (r = 0.999 5), 2.05-40.96 (r = 0.999 9), 3.65-72.96 (r = 0.999 9), 2.88-57.60 (r = 0. 999 8),13.25-264.96 mg x L(-1) (r = 0.999 6), respectively. The average recoveries (n = 6) of the six alkaloids were 101.6% (RSD 1.3%),102.5% (RSD 1.5%), 100.8% (RSD 1.9%),102. 6% (RSD 1.2%), 97.80% (RSD 1.3%), 99.01% (RSD 1.5%), respectively. The determined results demonstrate that there is a significant difference in the contents of six alkaloids and total alkaloids among the tested samples. The method is accurate, reliable and repeatable for simultaneous determination of gatrorrhizine, columbamine, epiberberine, coptisine, palmatine and berberine in C. chinensis.
Alkaloids ; isolation & purification ; Berberine ; analogs & derivatives ; isolation & purification ; pharmacology ; therapeutic use ; Berberine Alkaloids ; isolation & purification ; Chromatography, High Pressure Liquid ; methods ; Coptis ; chemistry ; Drugs, Chinese Herbal

OBJECTIVETo establish a RRLC-UV method for simultaneous determination of oxypeucedanin hydrate, bergapten, oxypeucedanin, imperatorin, cnidilin and oxypeucedanin in Angelica dahurica var. formosana.

METHODThe analysis was carried out on a Agilent Zorbax SB-C, (4.6 mm x 100 mm, 1.8 microm) column eluted with the mobile phases of methanol (A)-water (B) in gradient elution. The flow rate was 1.0 mL x min(-1), and the UV detection wavelength was set at 310 nm.

RESULTAll calibration curves of six coumarins showed good linearity regression within test ranges (r > 0.9998), and the overrall recoveries were in the range of 99.42%-101.4%, with RSD less than 2.4%.

CONCLUSIONThe method is simple, sensitive, reliable and reproducible which can be used for the quality study of Angelica dahurica var. formnosana.

Angelica ; chemistry ; Chromatography, High Pressure Liquid ; instrumentation ; methods ; Chromatography, Reverse-Phase ; methods ; Coumarins ; analysis ; Plant Extracts ; analysis ; isolation & purification

OBJECTIVE To compare the effect of intranasal corticosteroids-gelatin sponge and saline-gelatin sponge in the treatment of nasal adhesion.METHODS A total of 208 noses(170 patients) with nasal adhesions after receiving the endoscopic sinus surgeries from July,2012 to December,2015 were selected.All the cases were divided randomly into two groups:the treatment group and the control group.Both groups received separation treatments of the nasal adhesions.The intranasal corticosteroidsgelatin sponges were used as the separation material for the treatment group,while saline-gelatin sponges for the control group.The severity score and the time of treatment were evaluated as the indicators for curative efficacy of the treatment in each case.The differences of efficacy between two groups were analysed statistically.RESULTS The severity scores of the two groups after the treatment were both substantially lower than those before the treatment;The after-treatment severity scores of the treatment group were significantly lower than those of the control group;the times of treatment in the treatment group were substantially lower than those of the control group.The differences stayed for 3 months,and they became insignificant after 3 months.CONCLUSION The intranasal corticosteroids-gelatin sponge can be a quicker and more effective treatment of nasal adhesion in comparison with saline-gelatin sponge.

Objective:To study the effects of gantry acceleration limitations of a linear accelerator (linac) on the dosimetry of volumetric modulated arc therapy (VMAT) plans, machine efficiency, and dose verification result of VMAT plans and to explore the optimal selection of gantry motion models in the Pinnacle treatment planning system.Methods:Ten cases of nasopharyngeal carcinoma, non-small cell lung cancer, sigmoid adenocarcinoma with retroperitoneal lymph node metastasis, and invasive ductal carcinoma of the breast were each selected for this study. Then two models were set up in the Pinnacle v9.10 treatment planning system, namely the one allowing gantry acceleration and the one limiting gantry acceleration. The same field arrangement, optimized target parameters, and optimized weights of VMAT plans were adopted in the two models, in order to analyze the dosimetric variations in targets and organs at risk (OARs) and compare the differences in treatment time and gamma passing rates.Results:The treatment time of the enrolled patients under the model allowing gantry acceleration was significantly lower than that of the patients under the model limiting gantry acceleration was adopted ( t=-6.751, -0.209, -19.523, -28.999; P< 0.05) and decreased by 15.27%, 18.07%, 19.71%, and 28.75%, respectively. Meanwhile, the conformity and uniformity of target areas were affected, while there was no statistical significance in the gamma passing rates in the validation of VMAT plans ( P>0.05). For the cases of nasopharyngeal carcinoma (NPC), the maximum dose to brainstem PRV increased by 1.25%. For the cases of lung cancer, the maximum dose to the spinal cord and lung V20 increased by 1.19% and 1.21%, respectively, while lung V5 decreased by 1.21%. For the cases of sigmoid adenocarcinoma with retroperitoneal lymph node metastasis, the mean doses to bilateral kidneys, livers, small intestine, and colon all increased. For the cases of breast cancer, lung V10 on the opposite side of cancer increased by 1.66% and the mean dose to the lungs on the same side of cancer decreased by 7.45%. Conclusions:The model allowing gantry acceleration allows the treatment time to be significantly shortened and the treatment efficiency improved. Although this model had the shortcomings such as affecting the conformity and uniformity of target areas to a certain extent and increasing the doses to some OARs, clinical requirements for dosimetry were still met. Therefore, it is recommended to use the model allowing gantry acceleration in the Pinnacle planning system.

Objective: To explore effects of dendritic epidermal T cells (DETCs) and Vγ4 T lymphocytes on proliferation and differentiation of mice epidermal cells and the effects in wound healing of mice. Methods: (1) Six C57BL/6 male mice aged 8 weeks were collected and divided into control group and wound group according to random number table (the same grouping method below), with 3 mice in each group. A 4 cm long straight excision with full-thickness skin defect was cut on back of each mouse in wound group, while mice in control group received no treatment. On post injury day (PID) 3, mice in 2 groups were sacrificed, and skin within 5 mm from the wound margin on back of mice in wound group and normal skin on corresponding part of mice in control group were collected to make single cell suspensions. The percentage of Vγ4 T lymphocyte expressing interleukin-17A (IL-17A) and percentage of DETCs expressing insulin-like growth factor Ⅰ (IGF-Ⅰ) were detected by flow cytometer. (2) Ten C57BL/6 male mice aged 8 weeks were collected and divided into control group and Vγ4 T lymphocyte depletion group with 5 mice in each group. Mice in Vγ4 T lymphocyte depletion group were injected with 200 g Vγ4 T lymphocyte monoclonal neutralizing antibody of Armenian hamster anti-mouse intraperitoneally, and mice in control group were injected with the same amount of Armenian hamster Ig intraperitoneally. One hole with full-thickness skin defect was made on each side of spine of back of each mice. The wound healing was observed on PID 1-8, and percentage of remaining wound area was calculated. (3) Six C57BL/6 male mice aged 8 weeks were grouped and treated in the same way as in experiment (2), with 3 mice in each group. On PID 3, expressions of IL-17A and IGF-Ⅰ in epidermis on margin of wound were detected with Western blotting. (4) Thirty C57BL/6 male mice aged 3 days were sacrificed, and epidermal cells were extracted. The keratin 14 positive cell rate was examined by flow cytometer (the same detecting method below). (5) Another batch of mouse epidermal cells were collected and divided into control group, IGF-Ⅰ group, and IL-17A group, with 3 wells in each group (the same well number below). Cells in IGF-Ⅰ group and IL-17A group were added with 1 mL recombinant mouse IGF-Ⅰ and IL-17A with final mass concentration of 100 ng/mL respectively, while cells in control group were added with the same amount of sterile phosphate buffered saline (PBS). On post culture day (PCD) 5, keratin 14 negative cell rate was examined. Another batch of mouse epidermal cells were collected, grouped, and treated in the same way as aforementioned experiment, and keratin 10 positive cell rate was examined on PCD 10. (6) Another batch of mouse epidermal cells were collected and added with 4 mmol/L 5(6)-carboxyfluorescein diacetate N-succinimidyl ester (CFSE) solution, and divided into control 0 d group, control 7 d group, IGF-Ⅰ group, and IL-17A group. Cells in IGF-Ⅰ group and IL-17A group were treated in the same way as the corresponding groups in experiment (5), and cells in control 0 d group and control 7 d group were treated in the same way as the control group in experiment (5). The CFSE fluorescence peaks were examined on PCD 0 of control 0 d group and PCD 7 of the other 3 groups. (7) Another batch of mouse epidermal cells were collected and divided into control group and IGF-Ⅰ group. Cells in IGF-Ⅰ group were added with 1 mL recombinant mouse IGF-Ⅰ with final mass concentration of 100 ng/mL, and cells in control group were added with the same amount of sterile PBS. On PCD 5, cells were underwent keratin 14 staining and CFSE staining as aforementioned, and keratin 14 negative cell rate of CFSE positive cells was examined. Another batch of mouse epidermal cells were collected and divided into control group and IL-17A group. Cells in IL-17A group were added with 1 mL recombinant mouse IL-17A with final mass concentration of 100 ng/mL, and cells in control group were added with the same amount of sterile PBS. On PCD 5, keratin 14 negative cell rate of CFSE positive cells was examined. Data were processed with one-way analysis of variance and t test. Results: (1) On PID 3, percentage of DETC expressing IGF-Ⅰ in normal epidermis of control group was (9.9±0.8)%, significantly lower than (19.0±0.6)% of epidermis around margin of wound group (t=8.70, P<0.01); percentage of Vγ4 T lymphocyte expressing IL-17A in normal epidermis of control group was (0.123±0.024)%, significantly lower than (8.967±0.406)% of epidermis around margin of wound group (t=21.77, P<0.01). (2) On PID 1-4, there was obvious inflammatory reaction around wounds of mice in control group, and on PID 5-8, the wound area was still large. On PID 1-4, there was slight inflammatory reaction around wounds of mice in Vγ4 T lymphocyte depletion group, and on PID 5-8, the wound area was significantly reduced. On PID 3-7, percentages of residual wound area in Vγ4 T lymphocyte depletion group were significantly lower than those in control group (t=5.92, 5.74, 7.17, 5.38, 5.57, P<0.01), while percentages of residual wound area in two groups on PID 1, 2, 6 were similar (t=1.46, 3.17, 3.10, P>0.05). (3) On PID 3, compared with those in control group, expression of IL-17A and IGF-Ⅰ in epidermis around wound margin of mice in Vγ4 T lymphocyte depletion group was markedly decreased and increased respectively (t=8.47, 19.24, P<0.01). (4) The keratin 14 positive cell rate of mouse epidermal cells was 94.7%. (5) On PCD 5, the keratin 14 negative cell rate of mice in control group was markedly higher than that of IGF-Ⅰ group, while significantly lower than that of IL-17A group (t=7.25, 5.64, P<0.01). On PCD 10, the keratin 10 positive cell rate of mice in control group was significantly higher than that of IGF-Ⅰ group, while significantly lower than that of IL-17A group (t=3.99, 10.82, P<0.05 or P<0.01). (6) Compared with that of control 0 d group, CFSE fluorescence peaks of mouse epidermal cells in control 7 d group, IGF-Ⅰ group, and IL-17A group on PCD 7 shifted to the left. Compared with that of control 7 d group, CFSE fluorescence peaks of mouse epidermal cells in IGF-Ⅰ group and IL-17A group on PCD 7 shifted to the left. (7) On PCD 5, keratin 14 negative cell rate of CFSE positive cells of mice in control group was significantly higher than that in IGF-Ⅰ group (t=9.91, P<0.01), and keratin 14 negative cell rate of CFSE positive cells of mice in control group was markedly lower than that in IL-17A group (t=6.49, P<0.01). Conclusions In the process of wound healing, IGF-Ⅰ secreted by DETC can promote the proliferation of mouse keratin 14 positive epidermal cells and inhibit their terminal differentiation, while IL-17A secreted by Vγ4 T lymphocyte can promote the proliferation and terminal differentiation of mouse keratin 14 positive epidermal cells, thus both IGF-Ⅰ and IL-17A can affect wound healing.

Objective: To explore the effects of dendritic epidermal T cells (DETC) on proliferation and apoptosis of epidermal cells in wound margin of mice and its effects on wound healing. Methods: Twenty-eight healthy specific pathogen free (SPF) C57BL/6 wild-type (WT) male mice aged 8-12 weeks and 60 SPF T lymphocyte receptor δ-knockout (TCR δ^-/-) male mice aged 8-12 weeks were selected to conduct the following experiments. (1) Eight WT mice were selected to isolate epidermal cells and primarily culture DETC according to the random number table. Morphological observation and purity identification of DETC by flow cytometer were detected immediately after culture and on culture day (CD) 15 and 30, respectively. (2) According to the random number table, 5 WT mice and 5 TCR δ^-/- mice were selected and enrolled into WT control group and TCR δ^-/- group. Round full-thickness skin defect with diameter of 6 mm was made on the back of each mouse. The wound healing condition was observed immediately after injury and on post injury day (PID) 2, 4, 6, 8, 10, and the percentage of residual wound area was calculated. (3) Mice were selected to group and reproduce model of full-thickness skin defect as in experiment (2). On PID 3, the tissue of wound margin was collected for hematoxylin eosin staining, and the length of new epithelium was measured. (4) Mice were selected to group and reproduce model of full-thickness skin defect as in experiment (2). On PID 3, epidermal tissue of wound margin was collected to determine expression of proliferating cell nuclear antigen (PCNA) using Western blotting for evaluation of proliferation of epidermal cell. (5) Mice were selected to group and reproduce model of full-thickness skin defect as in experiment (2). On PID 3, epidermal tissue of wound margin was selected and digested into single-cell suspension, and apoptosis of cells was detected by flow cytometer. (6) Forty TCR δ^-/- mice were selected to carry out the same treatment as in experiments (2)-(5). According to the random number table, these mice were enrolled into TCR δ^-/- control group and TCR δ^-/-+ DETC group, with 5 mice in each group for each experiment. Round full-thickness skin defect was made on the back of each mouse. DETC in the number of 1×10⁵ (dissolution in 100 μL phosphate with buffer purity above 90%) were injected through multiple points of wound margin of mice in TCR δ^-/-+ DETC group immediately after injury, and equal volume of phosphate buffer was injected into mice of TCR δ^-/- control group with the same method as above. Data were processed with one-way analysis of variance for repeated measurement, t test, and Bonferroni correction. Results: (1) Along with the culture time elapse, the number of dendritic structures of DETC increased gradually. The percentage of T lymphocytes was 4.67% and 94.1% of these T lymphocytes were DETC. The purity of DETC on CD 15 was 18.50% and the purity of DETC on CD 30 was 98.70%. (2) Immediately after injury, the wound healing condition of mice in WT control group and TCR δ^-/- group was similar. The wound healing speed of mice in TCR δ^-/- group was slower than that in WT control group on PID 2-10. The percentages of residual wound area of mice in TCR δ^-/- group on PID 2, 4, 6, 8, and 10 were increased significantly compared with those in WT control group (t=3.492, 4.425, 4.170, 4.780, 7.318, P<0.01). (3) The length of new epithelium of mice in TCR δ^-/- group on PID 3 was (359 ± 15) μm, which was obviously shorter than that in WT control group [(462±26) μm, t=3.462, P<0.01]. (4) Immediately after injury, wound condition of mice in TCR δ^-/-+ DETC group and TCR δ^-/- control group was similar. Compared with TCR δ^-/-+ DETC group, the wound healing speed of mice in TCR δ^-/- control group were obviously slower on PID 2-10. The percentages of residual wound area of mice in TCR δ^-/-+ DETC group on PID 2, 4, 6, 8, and 10 were decreased significantly compared with those in TCR δ^-/- control group (t=2.308, 3.725, 2.698, 3.707, 6.093, P<0.05 or P<0.01). (5) On PID 3, the length of new epithelium of mice in TCR δ^-/-+ DETC group was (465±31) μm, which was obviously longer than that in TCR δ^-/- control group [(375±21) μm, t=2.390, P<0.05]. (6) On PID 3, PCNA expression of epidermal cell in wound margin of mice in TCR δ^-/- group was 1.25±0.04, which was obviously lower than that in WT control group (2.01±0.09, t=7.415, P<0.01). (7) On PID 3, PCNA expression of epidermal cell in wound margin of mice in TCR δ^-/-+ DETC group was 1.62±0.08, which was significantly higher than that in TCR δ^-/- control group (1.05±0.14, t=3.561, P<0.05). (8) On PID 3, apoptosis rate of epidermal cell in wound margin of mice in TCR δ^-/- group was (16.1±1.4)%, which was higher than that in WT control group [(8.1±0.6)%, t=5.363, P<0.01]. (9) On PID 3, apoptosis rate of epidermal cell in wound margin of mice in TCR δ^-/-+ DETC group was (11.4±1.0)%, which was obviously lower than that in TCR δ^-/- control group [(15.4±1.4)%, t=2.377, P<0.05]. Conclusions DETC participates in the process of wound healing though promoting the proliferation of epidermal cells in wound margin and inhibit the apoptosis of these cells.