AIM:To establish the endoplasmic reticulum stress ( ERS) cell model in vascular smooth muscle cells ( VSMCs) of Sprague-Dawley (SD) rats.METHODS:Under sterile condition, the coronary arteries were isolated from SD rats .The primary VSMCs were cultured by tissue-sticking method , and observed the basic morphological characteristics under optical microscope .The marker proteins of VSMCs including α-smooth muscle actin (α-SMA) and smooth muscle myosin heavy chain ( SM-MHC) were identified by immuno-fluorescence technique .VSMCs were treated with thapsigargin (0.5, 1 and 2 μmol/L) for 24 h, and the expression levels of binding immunoglobulin protein (BiP) and C/EBP homologus protein (CHOP), the marker molecules of ERS, were detected using Western blotting.RESULTS:VSMCs climbed out from coronary artery tissues after about six days , and the cells had a nice state and formed the VSMC-like typical peak valley.The results of immunofluorescence technique show that the marker proteins of VSMCs ,α-SMA and SM-MHC were expressed significantly .The results of Western blotting show that the protein expression levels of BiP and CHOP were increased by thapsigargin in a dose-dependent manner .CONCLUSION:VSMCs can be successfully cultured by tissue-sticking method and built the ERS model induced by thapsigargin .

Objective To investigate the value of differential diagnosis of 5-hydroxytryptamine,dopamine and their metabolic productions in cerebral spinal fluid(CSF) in patients with viral encephalitis and schizophrenia.Methods The CSF concentrations of DA,5-HT,5-hydroxyindole acetic acid(5-HIAA) and homovanillic acid(HVA) were assayed by high-performance liquid chromatography(HPLC) in the patients with viral encephalitis and meningoencephalitis(67 cases),schizophrenia(37 cases) and other diseases(21 cases).Results(1) Compared with case control group,the concentrations of 5-HT,5-HIAA and the ratios of 5-HIAA/5-HT,5-HT/DA were significantly lower in viral encephalitis and meningoencephalitis group(P

Objective : To understand the current situation and influencing factors of milk and dairy intake among primary and middle school students in Tongzhou District, Beijing, so as to provide the reference for the intervention of milk and dairy intake among primary and middle school students. Methods: The stratified cluster sampling method was used to randomly select the students from the third to sixth grades of primary school, three grades of junior middle school and three grades of senior high school from downtown and township of Tongzhou District as the survey objects. The demographic information, family data, and milk and dairy intake of a week before were collected. Taking the average daily intake of 300 g milk and dairy as the standard (the Chinese Dietary Guidelines 2016), the multivariate logistic regression model was used to analyze the influencing factors for reaching the standard of average daily milk and dairy intake among the students. Results: A total of 804 questionnaires were sent out, and 771 valid questionnaires were received, with a recovery rate of 95.90%. There were 321 primary school students, accounting for 41.63%; 228 junior high school students, accounting for 29.57%; and 222 high school students, accounting for 28.80%. The intake rate of milk and dairy products was 90.92%, and the adverse reaction rate was 10.12%. The daily intake rate was 36.71%. The median daily intake was 214.29 g, and the rate of average daily intake reaching the standard was 28.02%. The multivariate logistic regression results showed that primary school students (OR=1.672, 95%CI: 1.102-2.535), middle school students (OR=2.086, 95%CI: 1.349-3.225), overweight (OR=1.747, 95%CI: 1.131-2.700), obesity (OR=2.469, 95%CI: 1.698-3.591), and parents with bachelor's degree or above (OR=1.760, 95%CI: 1.022-3.029) were the influencing factors for reaching the standard of average daily milk and dairy intake among the students. Conclusions The average daily intake of milk and dairy products of primary and middle school students in Tongzhou District does not reach the recommended standard, and grade, body mass index and parents'education level were the influencing factors.

Objective To investigate blocking ascending branch of uterine arteries in operation of hysteromyomas rejecting by laparoscope.Methods 60 petients(except cervix myoma,intraligamentary myoma,submucosa myoma),randomly devided to reseach group(30 cases)and control group(30 cases).Blocking ascending branch of uterine arteries before rejecting hysteromyomas by laparoscope in reseach group,directly rejecting hysteromyomas by laparoscope in control group.The volumes of blood loss in operation,the duration of operation,postoperative recovery(Time of passing flatus,wound healing,length of stay)were compared between two groups.During the follow-up months,observing hysteromyoma recidivism and improvement of menstruation.Results All patient were rejected hysteromyomas by laparoscope.The volumes of blood loss in research group were less than that in control group(P0.05),All wounds were well healed in both groups,the rate of hypermenorrhea improvement was 100%(26/26)in research group,but That in control group was 80%(16/20),There was significant difference between two group.Conclusions Blocking ascending branch of uterine arteries may obviously decrease the volumes of blood loss in operation of hysteromyomas rejecting by laparoscope,It presents rapider convalescence,higher rate of symptom improvement and less complication operation.

Objective To evaluate the effects of dexmedetomidine with different-dose on the hemodynamics during anesthesia induction in patients undergoing off-pump coronary artery bypass grafting. Methods Sixty patients undergoing off-pump coronary artery bypass grafting were selected, with ASA grade Ⅱ - Ⅲ, NYHA cardiac functional grading Ⅱ - Ⅲ, and left ventricles ejection fraction >45%. The patients were divided into D1 group, D2 group and control group by table of random digit method with 20 cases each. In D1 group, intravenous infusion dexmedetomidine 0.3μg/kg was given for 20 min before anesthesia induction;in D2 group, intravenous infusion dexmedetomidine 0.6μg/kg was given for 20 min before anesthesia induction;in control group, intravenous infusion the same volume of 0.9% sodium chloride was given before anesthesia induction. The mean arterial pressure (MAP), heart rate, cardiac output and stroke volume variation (SVV) were recorded before infusion dexmedetomidine (T0), before anesthesia induction (T1), 3 min after anesthesia induction (T2), trachea cannula (T3) and 5 min after trachea cannula (T4). The adverse cardiovascular events and drug intervention were recorded during anesthesia induction. Results There were no statistical differences in MAP, heart rate, cardiac output and SVV at T0 among 3 groups (P>0.05). Compared with that in control group, the heart rate at T1, T2, T3 and T4 in D1 group were decreased, the MAP, heart rate, cardiac output and SVV at T1, T2, T3 and T4 in D2 group were decreased, and there were statistical differences (P<0.05). Compared with that at T0, the heart rate at T1, T2, T3 and T4 in D1 group were decreased, the MAP, heart rate, cardiac output and SVV at T3 in control group were increased, the MAP, heart rate, cardiac output and SVV at T1, T2, T3, T4 in D2 group were decreased, and there were statistical differences (P<0.05). Compared with that in D1 group, the MAP, heart rate, cardiac output and SVV at T2, T3 and T4 in D2 group were decreased, and there were statistical differences (P<0.05). The rates of adverse cardiovascular events in D1 group and D2 group were significantly lower than those in control group:35%(7/20) and 40%(8/20) vs. 95%(19/20), the rate of drug intervention in D1 group was significantly lower than that in control group and D2 group:10% (2/20) vs. 45% (9/20) and 35% (7/20), and there were statistical differences (P<0.05). Conclusions Dexmedetomidine (0.3 μg/kg) is beneficial for the stability of hemodynamics before anesthesia induction in patient undergoing off-pump coronary artery bypass grafting.

ObjectiveTo investigate the effect of controlled low perfusion pressure on expression of phosphor-Akt (p-Akt) and phosphor-ERK1/2 (p-ERK1/2) following spinal ischemia-reperfusion (1/R) in rabbits.MethodsThirty-six Japanese long-ear white rabbits aged 3 months weighing 2.0-2.5 kg were randomly divided into 3 groups ( n ＝ 12 each):group sham operation (group S) ; group spinal I/R (group I/R) and group controlled low perfusion pressure (group LP).Auricular artery and femoral artery were carnulated for proximal and distal BP monitoring.A 4F catheter with a balloon at the tip was inserted into abdominal aorta.The tip was positioned 1 cm below left renal artery.Spinal ischemia was induced by inflating the balloon until the distal MAP < 20 mm Hg,and maintained for 25 min.During the first 10 min of reperfusion the distal MAP was increased to 45-55 mm Hg by partially deflating the balloon.Then the balloon was fully deflated to allow complete reperfusion.The function of lower limbs was assessed with Tarlov score (0 ＝ no detectable movement,4 ＝ normal function) at 1,3,7,28 d of reperfusion.Au the animals were sacrificed at 1 d of reperfusion in each group.The lumbar segment L3-5 of the spinal cord was removed for microscopic examination and determination of expression of p-Akt and p-ERK1/2 by immuno-histochemistry) and detection of neuronal apoptosis in dorsal horn (by TUNEL).ResultsSpinal I/R significantly decreased Tarlov scores,and increased the number of apoptotic cells and expression of p-Akt and pERK1/2 in group I/R as compared with group S.Low perfusion pressure during the 10 min at the beginning of reperfusion significantly increased Tarlov scores,decreased apoptotic index and further increased p-Akt and pERK1/2 expression in group LP compared with group I/R.The histopathological damage in the spinal cord was attenuated in group LP.ConclusionControlled low pcrfusion pressure can reduce the spinal cord I/R injury by activating Akt and ERK1/2 and decreasing neuronal apoptosis.

Objective To comprehend the current difference of sequence variation of p24 coding region in gag gene and subtype distribution of HIV 1 in Henan province and Shanghai in China. Methods Plasma specimens were collected from 37 HIV 1 patients, of which 25 were from Henan province and 12 were from Shanghai. Most patients in Henan were infected by illegally donating blood while in Shanghai the major transmission routes were blood or blood product transfusion and sexuality. RNAs from plasma specimens were extracted and target gene were amplified by the method of RT PCR and Nested PCR. The sequences of p24 region, 693 nucleotides were determined, then phylogenetic analyses were performed. Results Subtyping showed that 83.8% (31/37) were B subtype. Of 25 Henan specimens, 23(92%)were B subtype and 2 were A subtype. Of 12 Shanghai specimens, 8(66.7%)were B subtype, 1 was A subtype, 2 were CRF01 -AE and 1 was CRF02 -AG. Comparing with the Consensus sequence (Consensus -B, from HIV database), nucleotide variation in B subtype of Henan was 1.6%～4.2%, with an average of 3.2%, while that of Shanghai was 2.0%～3.8%, with an average of 3.4%. No G to A hypermutation was observed in all variations. The median intrasubtype distance for B subtype in Henan and Shanghai was 2.9% and 3.5%, and the intersubtype distance between B subtype and other subtypes was 11.1%～12.5%. The two Consensus sequences of Henan and Shanghai B subtype obtained by CLUSTAL X were aligned to Consensus -B, both the substitution for predicted amino acid were 2.2%(5/231). Of all the variations,they shared the same three mutations: A14P、I91V and E180D. However, the disparity between the two Consensus sequences was not significant (0.9%). Phylogenetic analyses implied that many specimens were B subtype, and all the B subtype specimens from Henan and most of Shanghai B subtype specimens were close to the isolates in Thailand. Conclusion B subtype was the dominant HIV 1 isolate in Henan and Shanghai of China.The HIV 1 B subtype in Henan and Shanghai had coincident homology and shared some identical variations of amino acid.

BACKGROUND: Seed cells with good proliferation and enough amounts are need in reconstructing artificial salivary gland in vitro. However, adult stem cells are difficult to be isolated from normal submandibular gland.OBJECTIVE: To in vitro isolate submandibular gland stem/progenitor cells for cloning culture using animal models of damaged gland tissue.DESIGN, TIME AND SETTING: Cytological in vitro experiment was performed at the Gu.izhou Provincial Key Laboratory of Cell Tissue Engineering, Zunyi Medical College from March 2006 to January 2007.MATERIALS: A total of 10 male Sprague Dawley rats aged 8 weeks were supplied by the Animal Center, Third Military Medical University of Chinese PLA.METHODS: The model of tissue damaged submandibular gland in 10 rats was made by deligation. One week later, the gland tissue was obtained to harvest submandibular gland stem/progenitor cells by enzyme digestion in vitro. Following 10-14 days of primary culture, small round cells were collected, purified and subcultured for monoclonal culture.MAIN OUTCOME MEASURES: Immunocytochemical staining and immunofluorescence staining results were measured in submandibular gland stem/progenitor cells. Growth curve was drawn to analyze the proliferation of submandibular gland stem/progenitor cells in vitro.RESULTS: Cells expressing laminin showed stem cell characteristics. Positive expression of CD29 suggested high-adherent and high-proliferative stem cell properties. Positive expression of keratin-19 indicated epithelium-derived submandibular gland stem/progenitor cells. Growth curve was near to "S" shape, and in vitro culture and proliferation was active.CONCLUSION: Submandibular gland stem/progenitor cells had the characteristics of tissue stem cells. They might be as a kind of seed cells for tissue engineered artificial salivary gland in further research.

Objective To develop a phenylketonuria (PKU) screening method based on a compact disk (CD) type microfluidic chip capable of generating reciprocating flow within the microchannels that facilitate rapid DNA hybridization. Methods This microfluidic device consists of a two-layer structure: a polydimethylsiloxane (PDMS) top layer containing 12 DNA hybridization microchannels, and a bottom glass layer with immobilized hydrogel conjugated DNA arrays. The DNA arrays included R243Q, V245V and the blank control probes. When the CD device was spun, the PCR products were driven into the hybridization channel by centrifugal force. When the rotation of the CD device was stopped, capillary force pulled the PCR products solution to flow back to the channel. After the on-chip hybridization, the hybridization signals were captured on a fluorescence microscope. The specificity, detection limitation and reproducibility of this device were evaluated. Thirty DNA samples from pregnant women with suspected PKU were detected by this device.Then the results were compared with DNA sequencing results. Results With the compact disk type microfluidic chip, the hybridization time could be reduced to 15 min, sample consume could be as low as 1. 5 μl and the detection limitation was 0. 7 ng/μl. With the chip based method, samples of PKU patients and healthy controls were detected and the results were consistent with DNA sequencing results. Five different batches of chips and five micro-channels of each chip were selected to test one PKU patients with V245V mutation. All the results were positive, indicating good reproducibility. Four cases of V245V mutation and 1 case of R243Q mutation were found in 30 suspected PKU carried pregnant women. Conclusion The compact disk microfluidic device has advantages of simple, rapid and highly sensitive, thus is well suited to PKU screening.

AIM: To investigate the primary culture method for coronary artery smooth muscle cells (CASMCs), and to establish the endoplasmic reticulum stress ( ERS) model in CASMCs of SD rats.METHODS:CASMCs were cultured by tissue explant method .The morphological characteristics were observed under optical micro-scope.The marker proteins of CASMCs , including α-SMA and SM-MHC, were identified by immunofluorescence tech-nique.The protein expression levels of BiP and CHOP , the marker molecules of ERS, were determined by Western blot . RESULTS:The spindle-shaped CASMCs climbed out from the edge of coronary artery tissues after 6 d, and formed the typical hill and valleygrowth pattern of CASMCs at 9~10 d.The result of immunofluorescence technique showed that α-SMA and SM-MHC were positively expressed .The results of Western blot showed that the protein expression of BiP and CHOP in TG ( 1 and 2 μmol/L ) treatment groups was increased compared with control group .Compared with control group, the protein expression of BiP and CHOP was significantly increased after 1 μmol/L TG treatment for 24 and 48 h. CONCLUSION:CASMCs can be successfully cultured by tissue explant method .ERS model of CASMCs was established by 1 μmol/L TG treatment for 24 h.