BACKGROUND:Intervertebral disc degeneration is the basis of spinal degenerative diseases;however,there is no effective treatment. OBJECTIVE:To investigate whether sinomenine can inhibit interleukin-1β-induced apoptosis in nucleus pulposus cells and its molecular mechanism. METHODS:Rat nucleus pulposus cells were cultured in vitro by trypsin combined with type II collagenase digestion,and the cell growth curve was plotted.An appropriate sinomenine concentration was determined using the cell counting kit-8 kit.Nucleus pulposus cells were divided into control group,sinomenine group,interleukin-1β group,sinomenine+interleukin-1β group,zinc protoporphyrin group,zinc protoporphyrin+sinomenine group,zinc protoporphyrin+interleukin-1β group,and sinomenine+zinc protoporphyrin+interleukin-1β group.Proliferative activity,reactive oxygen species content,apoptosis rate,and heme oxygenase-1 expression in nucleus pulposus cells were detected. RESULTS AND CONCLUSION:The rat nucleus pulposus cells cultured in vitro were polygonal,triangular,and short wedge-shaped,and the cell growth showed an"S"curve.The cells grew slowly in the first 3 days of culture,rapidly in 4-6 days,and slowly again in 7-8 days.The cells then entered the"platform stage"where the number of cells no longer increased.The proliferative activity of myeloid cells showed no significant changes when the concentration of sinomenine was≤80 μmol/L(P>0.05).Interleukin-1β significantly reduced the proliferative activity of nucleus pulposus cells,increased the content of reactive oxygen species and led to apoptosis(P<0.01).Sinomenine intervention not only promoted heme oxygenase-1 expression(P<0.05)but also inhibited interleukin-1β-induced decrease in proliferative activity and increase in reactive oxygen species content and apoptosis rate in nucleus pulposus cells(P<0.05).These effects could be reversed by zinc protoporphyrin(P<0.01).

OBJECTIVE To explore the molecular mechanism of urolithin A inhibition of human hepatoma cells growth. METHODS Hepatoma Huh-7 cells were treated with different concentrations of urolithin A(Uro-A). The inhibition rate of Huh-7 cells survival was detected by CCK-8 assay and the IC50 was calculated. Cell proliferation was detected by colony formation assay and cell migration ability was assessed by cell wound healing experiment. Glucose uptake and lactate level in culture medium through colorimetry and the ATP production in cell through chemiluminescence method was analyzed. Western blotting was applied to detect protein expression levels of glucose transporter(GLUT1), key enzymes of glycolysis(HK2, PFKM, LDHA), p53, p-p38 and Bcl-2 after treatment with different concentrations of Uro-A. Flow cytometry and TUNEL method were used to detect apoptosis rate. RESULTS The results of CCK-8 showed that Uro-A significantly inhibited the proliferation of Huh-7 cells, and the IC50 was(48.54±1.21) μmol·L−1. The ability of clone formation and migration decreased after Uro-A treatment. Cellular glucose uptake and level of lactic acid and ATP production were down regulated in Huh-7 cells treated with Uro-A. The results showed that expression of glycolytic key proteins GLUT1, PKM2, LDHA and HK2 decreased. Western Blotting further research indicated that the p53 and p-p38 were activated, while the Bcl-2 was down-regulated. Flow cytometry data and TUNEL method revealed that the induction of apoptosis by Uro-A was remarkably increased. CONCLUSION These findings suggest that Uro-A can suppress Huh-7 cell proliferation and migration. The possible mechanism is the inhibition of glycolysis by p53, p-p38 and Bcl-2, which prevent cell growth and finally induce apoptosis.

BACKGROUND:Serum-specific biomarkers between normal healthy individuals and populations with developmental cervical canal stenosis(Qi deficiency and blood stasis syndrome)have not been fully defined. OBJECTIVE:To screen and identify the potential biomarkers of developmental cervical canal stenosis with Qi deficiency and blood stasis. METHODS:Serum samples were collected from nine patients with developmental cervical canal stenosis with Qi deficiency and blood stasis and eight healthy people.Differentially expressed proteins in serum were screened and identified using isotope relative labeling and absolute quantification combined with liquid chromatography tandem mass spectrometry.Western blot was used to verify some significant differentially expressed proteins. RESULTS AND CONCLUSION:A total of 61 differentially expressed proteins(P<0.05)were identified using tandem mass spectrometry techniques.Compared with the healthy normal population group,14 differentially expressed proteins such as complement component C1q receptor,apolipoprotein A4,and C-C motif chemokine ligand 18 were significantly upregulated,while 47 differentially expressed proteins such as myosin light chain 3,mitochondrial translation elongation factor,and nucleolar phosphoprotein 1 were significantly downregulated.The results of gene ontology enrichment analysis indicated that these differentially expressed proteins might participate in molecular functions such as regulation of chromosomal tissue,mitochondrial membrane tissue,and muscle system processes.Protein-protein interaction network analysis showed that 38 common differential proteins,including complement component C1q receptor,apolipoprotein A4,C-C motif chemokine ligand 18,myosin light chain 3,mitochondrial translation elongation factor,and nucleolar phosphoprotein 1,were located at functional network nodes between healthy normal individuals and those with developmental cervical canal stenosis(Qi deficiency and blood stasis syndrome),and were closely related to the local energy metabolism of the cervical spine,the production of cervical vertebral osteocytes,and the formation of osteoclasts.The main differentially expressed protein myosin light chain 3 was validated using western blot assay,and the validation results were consistent with the proteomic results.To conclude,the preliminary discovery of differentially expressed proteins in serum between healthy normal individuals and those with developmental cervical canal stenosis(Qi deficiency and blood stasis syndrome)through absolute quantitative technology combined with liquid chromatography tandem mass spectrometry technology suggests that myosin light chain 3 may be a specific serum marker for developmental cervical canal stenosis(Qi deficiency and blood stasis syndrome).

Osteoarthritis(OA)is the most common degenerative disease in clinical practice.Joint pain is the main manifestation of OA,and in severe cases,it can lead to joint deformity.The main pathological changes include destruction of articular cartilage,synovitis,thickening of subchondral bone,bone redundancy,degeneration of articular ligaments,and enlargement of articular periosteum.Clinicopathological changes and pain degrees in OA patients vary from person to person,and treatment options vary from person to person.Clinically,non-steroidal anti-inflammatory drugs are used to treat the pain of OA patients now,but the effect is not ideal,and long-term use of these drugs will bring obvious toxic and side effects.Traditional Chinese medicine obtains good results in relieving the pain of OA patients.Therefore,this paper reviews the research on traditional Chinese medicine treatment and pain mechanisms of OA,with a view to providing ideas and references for clinical relief of pain in OA patients.

OBJECTIVE: To explore the molecular pathological mechanism of liver metabolic disorder in severe spinal muscular atrophy (SMA). METHODS: The transgenic mice with type Ⅰ SMA (Smn^-/- SMN2^0tg/2tg) and littermate control mice (Smn^+/- SMN2^0tg/2tg) were observed for milk suckling behavior and body weight changes after birth. The mice with type Ⅰ SMA mice were given an intraperitoneal injection of 20% glucose solution or saline (15 μL/12 h), and their survival time was recorded. GO enrichment analysis was performed using the RNA-Seq data of the liver of type Ⅰ SMA and littermate control mice, and the results were verified using quantitative real-time PCR. Bisulfite sequencing was performed to examine CpG island methylation level in Fasn gene promoter region in the liver of the neonatal mice. RESULTS: The neonatal mice with type Ⅰ SMA showed normal milk suckling behavior but had lower body weight than the littermate control mice on the second day after birth. Intraperitoneal injection of glucose solution every 12 h significantly improved the median survival time of type Ⅰ SMA mice from 9±1.3 to 11± 1.5 days (P < 0.05). Analysis of the RNA-Seq data of the liver showed that the expression of the target genes of PPARα related to lipid metabolism and mitochondrial β oxidation were down-regulated in the liver of type Ⅰ SMA mice. Type Ⅰ SMA mice had higher methylation level of the Fasn promoter region in the liver than the littermate control mice (76.44% vs 58.67%). In primary cultures of hepatocytes from type Ⅰ SMA mice, treatment with 5-AzaC significantly up-regulated the expressions of the genes related to lipid metabolism by over 1 fold (P < 0.01). CONCLUSION Type Ⅰ SMA mice have liver metabolic disorder, and the down-regulation of the target genes of PPARα related to lipid and glucose metabolism due to persistent DNA methylation contributes to the progression of SMA.
Mice ; Animals ; PPAR alpha ; Liver Diseases ; Muscular Atrophy, Spinal/genetics* ; Mice, Transgenic ; Body Weight ; Glucose

Objective To study the clinical characteristics of Japanese encephalitis (JE) in 34 adult patients and to improve the level of diagnosis of this disease.Methods The clinical manifestations,laboratory results and radiological features of 34 adult patients with JE in our hospital from July 2017 to September 2017 were summarized and the progonsis was observed.The modified Rankin Scale (mRS) was used to evaluate the progonsis.Results Eighteen patients were males and 16 patients were females with the average age of (45.39 ± 16.34) years in 34 patients who were diagnosed as JE.The major clinical features of JE patients included fever (34,100％) with the average temperature of (39.4 ± 1.1) ℃ on admission,headache (26,76％),seizures (7,21％),decreased consciousness (25,74％) on day 2.6 ± 1.4 after the onset,respiratory failure (9,26％) on day 3.8 ± 1.6 after the onset.The major features of laboratory results included white blood cells increase (15,44％),blood hematocrit decrease (25,74％),eosinophil absolute value decrease (29,85％),cerebrospinal fluid pressure increase (12,35％),cerebrospinal fluid protein increase (27,79％),cerebrospinal fluid white blood cells increase (30,88％).Brain MRI scan of abnormal signal was found abnormal in up to 54％patients (14/26),involving the thalamus,basal ganglia,mesencephalon,temporal lobe,hippocampus and occipital lobe,especially in the area of bilateral thalamus and mesencephalon.The follow-up showed three cases were dead;mRS score was 0 in twenty-one cases,1 or 2 in five cases,3 or 4 in three cases,5 in two cases five-six months after onset;the sequelaes were cognitive impairment in nine patients and movement disorder in five patients.Conclusions The clinical symptoms of JE in adults are severe.The main clinical manifestations of JE are hyperthermia,disturbance of consciousness,seizures and respiratory failure,with characteristic imaging findings on brain MRI.JE is a disease with high mortality and severe long-term sequelae.

Objective To evaluate the clinical efficacy of phlegm and nourishing Yin combined with glucocorticoids in the treatment of refractory Mycoplasma pneumoniae pneumonia in children .Methods 134 children with Mycoplasma pneumonia were divided into control group and observation group according to the random number , 67 cases in each group .The control group were given hydrocortisone ,and the observation group received phlegm and nourishing Yin combined with glucocorticoid .After intervention ,the clinical data and the improvement of the clinical symptoms of the two groups were compared .Results There was no statistically significant difference in antipyretic time between the two groups(P>0.05).The time of cough relief,the absorption time of lung rales,the disappearance time of pleural effusion of the observation group were (3.61 ±1.13) d,(6.52 ±1.90) d and (7.02 ±2.10) d, respectively,which were significantly shorter than those of the control group [(4.89 ±2.18)d,t=2.920,P=0.021;(8.24 ±2.75)d,t=3.108,P=0.000;(9.26 ±2.38)d,t=5.027,P=0.000].There were no statistically signifi-cant differences in the total white blood cell count ,C-reactive protein and erythrocyte sedimentation rate between the two groups(all P>0.05).The percentage of lymphocytes in the observation group [(55.25 ±4.91)％] was signifi-cantly higher than that in the control group [(50.16 ±4.83)％,t=2.597,P=0.037].The effective treatment rate in the observation group was 97.01％,which was significantly higher than 83.58％in the control group (χ26.849,P=0.009).Conclusion Heat phlegm and nourishing Yin combined with glucocorticoid can not only effectively improve the patients'clinical symptoms , improve the proportion of lymphocyte , but also can improve the overall efficacy of children,it is worthy of clinical promotion .

Changes in metabolism after spinal cord injury cause many nutritional problems, leading to an increased incidence of dyslipidemia, diabetes and cardiovascular diseases. Unreasonable diet structure will promote the occurrence of metabolic abnormalities, induce secondary lesions, and increase mortality in patients with spinal cord injury at early stage. A comprehensive intervention including exercise, diet and dietary supplements is needed .

Objective To evaluate the application value of Smart Mask technology guiding micro-catheter in place for intracranial aneurysm embolization in endovascular interventional treatment. Methods The DSA data of 61 consecutive patients with unruptured intracranial aneurysm in the Department Neurosurgery,Changhai Hospital,the Second Military Medical University from August 5 to December 20,2016 were collected retrospectively. The conditions of the guildewire catheter reaching the aneurysms and the coil embolization in the Smart Mask technology group (n = 31)and conventional road map group (n = 30)were compared. The observation indexes included the differences in contrast agent dosage,exposure time,and operation time. Results The contrast agent dosage of the Smart Mask technology group and road map technology group were 192 ± 37 and 215 ± 40 ml respectively,the X-ray illumination quantity were 5913 ±1682 and 6975 ±2036 mGy respectively,and the operation time were 2. 62 ± 0. 29 h and 2. 94 ± 0. 35 h respectively. The differences were statistically significant (all P < 0. 05 ). Conclusion Using Smart Mask technology to assist endovascular embolization of intracranial aneurysms can effectively shorten the operation time,reduce the amount of X-ray radiation and contrast agent,and reduce the risk of operation.

OBJECTIVETo compare the effects of acupuncture on sphenopalatine ganglion and acupuncture on the common acupoints for life quality of patients with allergic rhinitis(AR).

METHODSEighty patients with AR,who were in accord with the inclusive criteria,were randomly divided into an observation group and a control group,40 cases in each one. Acupuncture on sphenopalatine ganglion was used in the observation group. The needle was inserted into the gap between zygoma and mandibular coronoid process about 55 mm. Acupuncture was adopted on the main acupoints,Yingxiang(LI 20),Yintang(GV 29) or Fengchi(GB 20) in the control group. The course was four weeks. Follow-up was applied one month after treatment. Rhinoconjunctivitis quality of life questionnaire (RQLQ),rhinitis symptoms scale and visual analogue scale(VAS) were evaluated at different time points before and after treatment,and follow-up was implemented to know the recurrence situation,satisfactory degree and adverse reaction.

RESULTS(1) RQLQ scores:along with treatment,the RQLQ scores were gradually apparently decreased in the two groups(<0.01),and the reducing trend from the first week to the second week of the observation group was more obvious than that of the control group. The differences of the RQLQ scores at all timepoints after treatment between the two groups were not statistically significant(all>0.05). The interaction of the time factor and the group factor had statistical significance(<0.01). (2) Rhinitis symptoms scores:along with treatment,the scores presented decreasing trend in the two groups(<0.01). The scores of the two groups after treatment and the interaction of the time factor and the group factor were not statistically different(both>0.05). (3)VAS scores:the VAS scores after treatment were lower than those before treatment(both<0.01). The differences before and after treatment were statistically significant(<0.01),with more change in the observation group(<0.05). (4)There was no statistical significance about the number of recurrence days between the two groups(>0.05). (5) Above 80 percent patients were content with the therapeutic method in each group,with no statistical difference(>0.05). (6)The method of the observation group spent less time. (7) Two patients with light adverse reaction came up in the observation group,but no special treatment was needed.

CONCLUSIONSAcupuncture on sphenopalatine ganglion acquires more obvious short-term effect than conventional acupuncture. It spends less time to relieve symptoms and improves life quality.