Components of free fatty acids (FFA) were examined by reverse high performance liquid chromatography in 25 patients with type 2 diabetes and 25 control subjects.The changes in components of FFA were observed before treatment, at 3 months and 1 year after treatment with pioglitazone in type 2 diabetic patients.The serum lauric acid, myristic acid and stearic acid levels were much higher in the diabetic patients than those in control subjects.Pioglitazone could decrease the levels of these fatty acids in the patients with type 2 diabetes mellitus.

BACKGROUND: Strontium-doped calcium polyphosphate (SCPP) is a new type of bone repair materials with good biocompatibility and controlled degradation. The preliminary studies of our group indicate their role in promoting angiogenesis,but its mechanism is unclear.OBJECTIVE: By co-culturing osteoblasts ROS17/2.8 with SCPP in vitro to observe cell proliferation and the secretion of vascular endothelial growth factor (VEGF).DESIGN, TIME AND SETTING: A contrast study was performed at the Laboratory of Tissue Engineering of Sichuan University from October 2008 to June 2009.MATERIALS: A series of calcium polyphosphate (CPP) respectively containing 0%, 1 %, 2%, 5%, 8%, and 10% Sr~(2+) were prepared. ROS17/2.8 osteoblastic cell strain was provided by Laboratory of Transplantation Immunity and Transplantation Engineering, West China Hospital, Sichuan University.METHODS: ①Preparation of cell scaffold complexes: The materials were placed in 24-well plates, then 300 μL cell suspension with a concentration of 2×10~7 cells/Lwas inoculated into each hole. These complexes were cultured for 14 days and the liquid was changed every two days. ②These complexes were measured by MTT assay to observe the proliferation of osteoblasts on the 1~(st), 3~(rd), 5~(th), 7~(th), 10~(th) and 14~(th) days, respectively. ③ The centrifugal supernatant of the complex cultured for seven days was measured by ELISA assay to check the secretion of VEGF.MAIN OUTCOME MEASURES: The proliferation of osteoblastic cells on SCPP and CPP was observed. The amount of VEGF protein secreting from osteoblastic cells was detected.RESULTS: The results of MTT showed that, compared with the CPP group, SCPP groups could promote the proliferation of osteoblasts, and 8% SCPP group was the best; ELISA results showed that, compared with the CPP group, SCPP groups could increase the amount of VEGF protein secretion, of which the promoting role of 8% SCPP was the most obvious (P < 0.05).CONCLUSION: When cultured with osteoblasts, SCPP can promote cell proliferation, and can significantly increase the secretion of VEGF; moreover, 8% SCPP is the best, which reveals a certain mechanism of its promoting angiogenesis.

Objective To evaluate the feasibility of Mycobacteria growth indicator tube ( MGIT ) liquid culture combined with rifampin resistance test real-time PCR ( Xpert MTB/RIF test) in the diagnosis of tuberculosis.Methods 652 cases of suspected pulmonary tuberculosis from October 2014 to March 2015 in Quzhou People′s Hospital were enrolled. The morning sputum samples were collected for acid-fast staining, L?wenstein-Jensen ( L-J) culture, BACTEC MGIT culture and Xpert MTB/RIF test.Samples with positive results of MGIT liquid culture were subjected to strain identification and drug sensitivity test with fluid method.Methodological comparison between four methods was made and the performance of MGIT liquid culture combined with Xpert MTB/RIF test in the rapid diagnosis and drug resistance detection in Mycobacterium tuberculosis was evaluated by chi-square test. Results Among the samples from 399 confirmed tuberculosis patients, the diagnostic sensitivity of the 4 methods ( acid-fast staining, L-J culture, MGIT culture and Xpert MTB/RIF test) were 17.0%(68/399), 23.8%(95/399), 37.8% (151/399) and 37.3%(149/399) respectively.The sensitivity and specificity of MGIT culture combined with Xpert MTB/RIF test was 39.8%(159/399) and 94.8%(240/253).The sensitivity of MGIT culture combined with Xpert MTB/RIF test was significantly higher than acid-fast staining (χ2 =50.9, P<0.01 ) and L-J culture (χ2 =23.7,P<0.01).The average detection time of MGIT culture was 7.5 days ( smear positive and Xpert MTB/RIF positive), 13.4 days (smear negative and Xpert MTB/RIF positive) and 16.9 days ( smear negative and Xpert MTB/RIF negative) .The sensitivity and specificity of Xpert MTB/RIF test in rifampin resistance detection were 9/9 and 97.3% ( 129/132 ) respectively.The average detection time of fluid method for drug sensitivity test was 8.3 days.Conclusions MGIT liquid culture combined with Xpert MTB/RIF test can detect Mycobacterium tuberculosis and the drug sensitivity rapidly.The method is highly sensitive and specific.It is important for clinical diagnosis and treatment of tuberculosis.

Objective To observe the expression of the unfolded protein response especially the inositol-requiring enzyme-1 (IREl)-Xbp1 signaling pathway, and the change trend of osteogenic markers after inhibition of IREl expression through siRNA interference in osteoblasts exposed to fluoride. Methods Proliferation activity of MC3T3-E1 cells was detected by CCK-8 assay, and 0.0, 0.1, 1.0, 2.0, 8.0, 16.0, 20.0, 32.0, 64.0 mg/L groups were set up. Then representative doses of low, medium and high fluoride (2.0, 8.0, 20.0 mg/L) were selected to treat MC3T3-E1 cells and the expression of the unfolded protein response related genes and osteogenic markers [alkaline phosphatase (ALP), osteocalcin (OCN), Runx2, osterix, binding immunoglobulin protein (Bip), protein kinase-like endoplasmin reticulum kinase ( PERK ) , activated transcription factor 6 (ATF6), Xbp1] was detected by Real-time PCR. MC3T3-E1 cells were transfected with IRE1 siRNA and then exposed to fluoride, and the expression of IRE1 signaling pathway and osteogenic markers was detected by Western blotting and real-time PCR. Results CCK-8 results showed the bidirectional effect of fluoride on the activity of osteoblasts. Compared with the 0.0 mg/L group [1.00 ± 0.01 (d 1), 1.00 ± 0.02 (d 3), 1.00 ± 0.08 (d 7)], the osteoblast activity was significantly enhanced at 2.0 mg/L [1.11 ± 0.02 (d 1), 1.29 ± 0.02 (d 3)], 8.0 mg/L [1.16 ± 0.02 (d 1), 1.44 ± 0.03 (d 3), all P<0.05], while 20.0 mg/L inhibited cell activity [0.83 ± 0.01 (d 1), 0.81 ± 0.01 (d 3), 0.96 ± 0.04 (d 7), all P< 0.05]. Compared with the 0.0 mg/L group [6.86 ± 2.13 (ALP), 4.58 ± 1.52 (OCN), 2.65 ± 0.38 (Runx2), 12.48 ± 3.96 (osterix)], 2.0 mg/L significantly induced the expression of intracellular ALP (12.80 ± 3.62), Runx2 (6.61 ± 0.48) and osterix (21.42 ± 1.56), and the differences were statistically significant (all P< 0.05), while 20.0 mg/L inhibited the expression of ALP (0.88 ± 0.17), OCN (0.16 ± 0.05) and osterix (1.35 ± 0.51), and the differences were statistically significant (all P<0.05). Compared with the 0.0 mg/L group [1.36 ± 0.58 (IRE1), 0.96 ± 0.45 (Xbp1)], the expression of endoplasmic reticulum stress related genes IRE1 [14.84 ± 2.57 (2.0 mg/L), 4.10 ± 0.52 (8.0 mg/L), 5.30 ± 0.63 (20.0 mg/L)] and Xbp1 [2.62 ± 0.66 (2.0 mg/L), 1.97 ± 0.47 (20.0 mg/L)] were significantly increased in the corresponding fluoride groups (all P<0.05). After IRE1 gene knockout, compared with the control group [gene:3.25 ± 0.48 (OCN), 5.62 ± 1.86 (Runx2), 2.67 ± 0.35 (ALP); protein: 0.16 ± 0.03 (OCN), 0.34 ± 0.27 (ALP)], the gene expression of OCN [0.63 ± 0.46 (2.0 mg/L), 0.81 ± 0.36 (8.0 mg/L), 0.62 ± 0.31 (20.0 mg/L)], Runx2 [0.18 ± 0.03 (2.0 mg/L), 0.12 ± 0.01 (8.0 mg/L), 1.09 ± 0.33 (20.0 mg/L)] and ALP [1.01 ± 0.12 (8.0 mg/L), 0.38 ± 0.09 (20.0 mg/L)] in the corresponding fluoride groups were significantly decreased (all P < 0.05), protein expression of OCN [0.06 ± 0.02 (2.0 mg/L), 0.06 ± 0.02 (8.0 mg/L), 0.07 ± 0.03 (20.0 mg/L)], and ALP [0.02 ± 0.01 (8.0 mg/L), 0 (20.0 mg/L)] were significantly decreased (all P< 0.05). Conclusion Unfolded protein response is observed under different doses of fluoride in osteoblasts, and IRE1 gene knockout has inhibited the expression of ALP, OCN, osterix and Runx2 in osteoblasts induced by fluoride, which suggests that IRE1 signaling pathway may play a key role in the differentiation of osteoblasts exposed to fluoride.

Objective To use the matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) technique for detecting the mutation gene of neonatal non-syndromic hereditary hearing impairment gene in Guangxi and to investigate its effectiveness and feasibility in clinical application.Methods A total of 7 100 newborns were performed the hearing preliminary screening and secondary screening by adopting AABR.The genomic DNA was extracted by the heel blood spot.Twenty mutation characteristics of 4 deaf predisposing genes were detected by MALDI-TOF-MS.Results The pass rate of hearing screening in 7 100 newborns was 97.11％ (6 895/7 100),the positive rate of neonatal gene mutation was 3.54％ (251/7 100),in which the GJB2 gene mutation was in 131 cases,the carrying rate was 1.84％,235delC heterozygous mutation was in 108 cases.SLC26A4 gene mutation was in 93 cases,which dominated by 1229C＞T heterozygous mutation and IVS7-2A＞G heterozygous mutation,mtDNA12SRNA gene mutation was in 16 cases and GJB3 gene mutation was in 11 cases.Conclusion Adopting the MALDI-TOF-MS screening technique can increase the detection rate of hot point mutation in common deaf related genes and discover neonatal genetic NSHI from molecular level and provides the corresponding geneticconsulting guidance for early finding and predicting deaf occurrence,and formulating the interventional measures.

Objective:To observe the effect of liver-soothing and mind-regulating acupuncture method on the resting-state electroencephalography (EEG) in rats with post-traumatic stress disorder (PTSD),and to provide evidence for the effect mechanism study and clinical application of acupuncture intervention for PTSD.Methods:Sixty Sprague-Dawley (SD) rats were randomly divided into a blank control group,a model group,a grasping group,a paroxetine group and an acupuncture group,with 12 rats in each group.Except for rats in the blank control group,rats in the other groups were subjected to preparing the PTSD models using 'incarceration plus electric shock' method.After interventions,changes in rat behavior of each group were observed;changes in resting-state EEG were collected and analyzed with multichannel EEG acquisition and analysis system,and image analysis and statistical processing were performed.Results:Compared with the blank control group,the average escape latency in the model group was significantly longer (P＜0.05),and the times of crossing the platform and the effective areas were all significantly reduced (P＜0.01).Compared with the grasping group,the average escape latencies in the paroxetine group and acupuncture group were significantly shortened (P＜0.05),and the times of crossing the platform and the effective areas were all significantly increased (P＜0.05).There were no significant differences in the average escape latency,the times of crossing the platform and the effective areas between the acupuncture group and paroxetine group (all P＞0.05).Compared with the blank control group,the α-wave power spectrum value in the model group was significantly decreased,and the power spectrum values of β-wave,δ-wave and a-wave were significantly increased (all P＜0.01);compared with the grasping group,α-wave power spectrum values in the paroxetine group and acupuncture group were significantly increased (both P＜0.01),and the power spectrum values of β-wave,δ-wave and a-wave were decreased significantly (all P＜0.01).The power spectrum values of α-wave,β-wave,δ-wave and (e)-wave of rats in the acupuncture group were not significantly different from those in the paroxetine group (all P＞0.05).Conclusion:Liver-soothing and mind-regulating acupuncture method can significantly improve the abnormal EEG activity in PTSD rats,which may be one mechanism of liver-soothing and mind-regulating acupuncture method in effectively affecting the brain function in PTSD rats.

This study investigated the protective effect of chrysin on hepatic fibrosis by regulating AMP-activated kinase (AMPK)-NOD-like receptor protein 3 (NLRP3) mediated pyroptosis pathway. The hepatic fibrosis model of mice was established by thioacetamide (TAA) in vivo. Except the control and chrysin alone groups, the mice were injected intraperitoneally with TAA at 100 mg·kg^-1, three times per week for the first week. From the 2^nd to 5^th week, mice were injected intraperitoneally with TAA at 200 mg·kg^-1, three times per week for the next 4 weeks. Chrysin groups were intragastrically administrated once per day to 5^th week. The histopathological changes were detected by HE and Masson staining. The levels of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were assessed by the kits. All animal experiments were approved by the Medical Ethics Committee of Affiliated Zhongshan Hospital of Dalian University (DWLL2019060). LX-2 cells were stimulated by (transforming growth factor-β, TGF-β) in vitro. The protein expressions of AMPKα, p-AMPKα, NLRP3, cysteinyl aspartate specific proteinase-1 (caspase-1), gasdermin D (GSDMD) were detected by Western blot, and the mRNA levels of collagen-Ι, α-smooth muscle actin (α-SMA), interleukin-1β (IL-1β), IL-18, caspase-1, GSDMD were analysis by reverse transcription-polymerase chain reaction (RT-PCR). Chrysin attenuated the increases in serum AST and ALT levels in the TAA group, while significantly improved the changes of liver morphology, reduced liver tissue inflammatory cell infiltration and inhibited collagens deposition. Compared with TAA group, chrysin effectively activated AMPKα phosphorylation and inhibited hepatic NLRP3 inflammasome activation. Additionally, the protein expressions and mRNA levels of IL-1β, IL-18, caspase-1 and GSDMD in chrysin groups were decreased. Chrysin inhibited the expressions of collagen-Ι and α-SMA, enhanced the phosphorylation of AMPKα, and decreased the expressions of NLRP3 and GSDMD. Therefore, chrysin may inhibit inflammatory injury and pyroptosis possibly by activating AMPK and inhibiting NLRP3 inflammasome to alleviate hepatic fibrosis.

Objective: To observe the clinical effect of acupoint sticking therapy with Mian Tan Gao (facial paralysis paste) plus electroacupuncture (EA) for treating peripheral facial paralysis and its influence on patients' facial nerve functions, facial disability index and clinical symptoms and signs. Methods: A total of 96 peripheral facial paralysis patients were allocated into an observation group, a medicine group and an EA group by simple randomization, with 32 cases in each group. Patients in the medicine group were treated with oral mecobalamine and prednisone acetate; patients in the EA group were treated with EA on the basis of the medicine treatment; while patients in the observation group were treated with acupoint sticking therapy with Mian Tan Gao (facial paralysis paste) plus EA. After 4-week treatment, the clinical efficacy, the adverse events, and the scores of House-Brackmann (H-B) facial nerve function grading scale, visual analog scale (VAS), clinical symptoms and signs, and facial disability index (FDI) were compared. Results: After 4-week treatment, the total effective rate was 96.9％ in the observation group, higher than 68.7％ in the medicine group and 75.0％ in the EA group (both P<0.05). After 4-week treatment, the scores of H-B grading scale, VAS and clinical symptoms and signs in the three groups dropped significantly compared with those before treatment, and the scores in the observation group were lower than those in the medicine group and EA group (all P<0.05). After 4-week treatment, the facial disability index-physical function (FDIP) in the FDI in the three groups increased significantly, with a higher value in the observation group compared with that in the medicine group and EA group (both P<0.05). The facial disability index-social function (FDIS) in the FDI dropped significantly, with a lower score in the observation group compared with that in the medicine group and EA group (both P<0.05). However, the comparisons of the items above between the medicine group and the EA group showed no statistical significance (all P>0.05). The between-group comparison of the adverse event across the three groups showed no statistical significance (P>0.05). Conclusion: Acupoint sticking therapy with Mian Tan Gao (facial paralysis paste) plus EA can decrease H-B grade, reduce pain severity and improve clinical symptoms and signs as well as the facial disability condition in peripheral facial paralysis patients. This method produced more significant efficacy compared with oral medicine and medicine plus EA.

AIM: To clarify the effects of gastrin on t he expression of cyclooxygenase (COX) and several growth factors in rat gastric mu cosa. METHODS: Male Sprague Dawley rats were fasted for 24 hours and s ubcutaneously injected with saline or gastrin 17 at doses of 1 ?g/kg, 10 ?g/kg and 100 ?g/kg, respectively. The expression of COX-1, COX-2, heparin-binding e p idermal growth factor-like growth factor (HB-EGF) and hepatocyte growth factor ( HGF) in the gastric mucosa were examined using Western blotting and immunohistoc hemical staining. Effects of a potent gastrin receptor antagonist YM022 on the e xpression of COX-1, COX-2, HB-EGF and HGF in gastric mucosa were also evaluated. RESULTS: Gastrin dose-dependently increased the expression of C OX-2 and HB-EGF in rat gastric mucosa while the expression of COX-1 and HGF did not change significantly after treatment with gastrin. However, pretreatment wit h YM022 dose-dependently abolished the up-regulation of COX-2 and HB-EGF express ion induced by gastrin. CONCLUSIONS: This study demonstrates that gastrin up-regulates C OX-2 and HB-EGF expression in rat gastric mucosa, indicating that COX-2 and HB-E GF are involved in pathogenesis of the gastrin-related gastric mucosal hyperplas ia and carcinoma of stomach.

AIM: To clarify the effects of specific and non-specific cyclooxygenase-2 (COX-2) inhibitors on gastric epithelial cell proliferating and gastric healing following acid-induced damage. METHODS: Male Sprague-Dawley rats were given 1 mL of 0.6 mol/L hydrochloric acid (HCl) into the stomach. Ten minutes after the administration of the acid,the animals were given NS-398 (COX-2 inhibitor) or indomethacin. Levels of COX-1 and COX-2 in the gastric mucosa before and after HCl-administration were analyzed using western blotting and immunohistochemical staining. Proliferating cell nuclear antigen (PCNA) was detected using immunohistochemistry for epithelial cell proliferation. Gastric lesion index (LI) was assessed using planimetry. RESULTS: Expression of COX-2 was enhanced mainly in surface epithelial cells and neck cells following HCl-administration. At 24 h following acid administration,PCNA labeling index (PCNA-LI) was (22.72?4.33) % and (21.98?5.18) % in the groups treated with 40 mg/kg of NS-398 and indomethacin respectively,which was significantly lower than that in the control group [ (34.46?3.61) %,P