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1.
China Pharmacy ; (12): 560-564, 2023.
Article in Chinese | WPRIM | ID: wpr-964765

ABSTRACT

OBJECTIVE To establish the quality control method of Flueggea suffruticosa. METHODS The microscopical identification and thin layer chromatography (TLC) of F. suffruticosa were carried out, and the mass fractions of moisture, total ash, acid-insoluble ash and alcohol-soluble extracts in F. suffruticosa were measured based on the 2020 version of Chinese Pharmacopoeia (part Ⅳ). The content of securinine in medicinal materials was determined by high performance liquid chromatography. RESULTS The powder of F. suffruticosa was gray-green, with obvious microscopic characteristics such as pores, pollen grains, calcium oxalate cluster crystals, ducts. The results of TLC identification showed that in the chromatograms of 16 batches of medicinal samples, the same color spots were found on the corresponding positions of the chromatograms of securinine, rutin, quercetin and F. suffruticosa control. The average mass fractions of moisture, total ash, acid-insoluble ash and alcohol- soluble extracts in 16 batches of medicinal materials were 9.26%, 6.96%, 1.17% and 28.89%, respectively. The injection volume of securinine in the range of 0.052 4-0.524 0 µg had a good linear relationship with the peak area (R2=0.999 8). RSDs of precision, repeatability and stability (24 h) test were all less than 3% (n=6 or 7). The average recovery of sample was 97.47%, RSD was 1.63% (n=6). The content of securinine in 16 batches of medicinal materials was 1.003-6.872 mg/g. CONCLUSIONS The quality control method of F. suffruticosa is established, and the mass fractions of moisture, total ash and acid-insoluble ash should not exceed 12.0%, 9.0% and 2.0%, respectively; the alcohol-soluble extract should not be less than 20%, and the content of securinine should not be less than 1.00 mg/g.

2.
China Pharmacy ; (12): 861-866, 2022.
Article in Chinese | WPRIM | ID: wpr-923194

ABSTRACT

OBJECTIV E To optimize the s alt-processing technology of Rosa laevigata ,and to study high performance liquid chromatography(HPLC)fingerprints and chromaticity values of R. laevigata before and after salt-processing. METHODS The comprehensive scoring method was adopted to optimize the salt-processing technology of R. laevigata using appearance character , moisture and polysaccharide content as index. Fingerprints were established by HPLC method before and after salt-processing ,and chromaticity values (L*,a*,b*)of the powder before and after salt-processing were determined. The multivariate statistical analysis was carried out for raw product and salt-processing product of R. laevigata by using common peak areas and chromaticity values as index. RESULTS The optimal salt-processing technology of R. laevigata was to mix it with appropriate amount of salt water ,place them in the preheated frying wok at 140 ℃,fry them for 25 min,and rotate frying wok 20 times/min. Ten common peaks were calibrated by HPLC fingerprints before and after salt-processing ,and 3 components were identified ,such as gallic acid ,catechin and ellagic acid. The chromaticity values L*,b* and E* changed significantly after salt-processing. The multivariate statistical analysis method could distinguish raw products and salt-processing products into two categories ,in which peaks 1,5,6 and 10 and chromaticity values b* and E* were important characteristic factors. CONCLUSIONS The optimized salt-processing technology is stable and reliable ,and the established fingerprint has good repeatability and stability. Fingerprint and chromaticity values combined with multivariate statistical analysis can provide reference for the identification and quality analysis of R. laevigata before and after salt-processing.

3.
Clinical and Experimental Otorhinolaryngology ; : 109-114, 2017.
Article in English | WPRIM | ID: wpr-66653

ABSTRACT

OBJECTIVES: To assess the performance of contrast-enhanced ultrasound (CEUS) intended to differentiate hyperplastic from malignant neck lymph nodes in an animal model. METHODS: Twenty-four New Zealand rabbits were randomly divided into two groups: neck lymph node metastasis group (12 rabbits) and reactive hyperplastic lymph node group (12 rabbits). Tongue VX2 carcinoma with cervical lymph node metastasis was induced in 12 rabbits by injecting VX2 carcinoma suspension into the left tongue submucosa. Hyperplastic neck lymph nodes were induced by injecting egg yolk in the submandibular region of the rabbits in hyperplastic group. CEUS were performed in both groups before and after intravenous administration of SonoVue. The site, number, echogenicity, longitudinal and transverse nodal dimensions, patterns of enhancement of the neck lymph nodes were observed and recorded. RESULTS: In both groups only one lymph node was found in the left (tumor) side of the neck. CEUS found 12 of 12 metastatic lymph nodes in metastasis group, and diagnosed 11 of 12 lymph nodes as metastatic. Histopathologic analysis revealed metastatic lesions in all 12 rabbits, each with one lymph node, and all 12 lymph nodes in hyperplastic group is inflammation lymph nodes. All 12 cases in the hyperplastic group showed centripetal homogeneous enhancement while in the metastasis group one case showed centripetal homogeneous enhancement, three cases showed centrifugal heterogeneous enhancement, and eight cases showed diffused heterogeneous enhancement. Only one lymph node was dissected on the left side of the neck in each rabbit in both groups. CONCLUSION: CEUS can play a role in discriminating metastatic from hyperplastic lymph nodes in head and neck carcinoma.


Subject(s)
Animals , Rabbits , Administration, Intravenous , Egg Yolk , Head , Head and Neck Neoplasms , Inflammation , Lymph Nodes , Models, Animal , Neck , Neoplasm Metastasis , Tongue , Ultrasonography
4.
Journal of Clinical Otorhinolaryngology Head and Neck Surgery ; (24): 1386-1389, 2015.
Article in Chinese | WPRIM | ID: wpr-749158

ABSTRACT

To determine appropriate surgical methods and flaps to apply plastic surgery of facial defects. Several plastic methods were introduced progressively to eight cases. From simple to complex, we discussed the direct suture, relaxation suture, Z-flap, flap-footed, combined or multiple flaps, and free flap method to decrease the tension in wounds. The skin and flaps were successful in all eight cases and healed the wounds. It is important to choose appropriate surgical techniques and flaps to repair facial injuries.


Subject(s)
Humans , Face , General Surgery , Facial Injuries , General Surgery , Free Tissue Flaps , Plastic Surgery Procedures , Skin , Wound Healing
5.
Journal of Clinical Otorhinolaryngology Head and Neck Surgery ; (24): 1121-1125, 2015.
Article in Chinese | WPRIM | ID: wpr-747256

ABSTRACT

OBJECTIVE@#To investigate the effect of dxamethasone (DEX) on the expression of Tregs in allergic rhinitis (AR) mice, and explore the mechanism of glucocorticoid in the treatment of AR.@*METHOD@#AR murine model was established by sensitization and challenge with OVA, besides intervention treatment with DEX was carried out in AR model. The behavior observation was used to evaluate the improvement effect of DEX on AR symptoms. The morphological characteristics of nasal tissues were observed by HE staining after fixation and decalcification. The mononuclear cells were obtained by grinding spleens, and the total RNA was extracted for reverse transcriptase polymerase chain reaction to investigate the level of mRNA expression of Foxp3. The changes of CD4+ Foxp3+ Tcells in spleen of mice were analyzed by flow cytometry.@*RESULT@#BALB/c mice received OVA sensitization followed by OVA intranasal challenge, the frequencies of sneezing and nose-scratching increased significantly in AR group (44. 50 ± 5. 61 and 72. 94 ± 8. 76) compared with control group (12. 68 ± 1. 87 and 26. 76 ± .2. 89), P<0. 01; The frequencies decreased significantly in DEX group (26. 04 ± 3. 93 and 56. 79 ± 5. 64), P< 0. 05 compared with AR group. The continuity of nasal mucosa ciliated columnar epithelium in AR group was destroyed and appeared to be repaired in DEX group. Inflammatory cells infiltration was also markedly decreased by DEX treatment. The proportion of CD4+ Foxp3+ T cells in AR group (3. 89 ± 0. 39)% decreased, P<0. 01 vs control group (4. 63 ± 0. 15) %. DEX treatment induced production of Tregs (6. 89 ± 0. 49)%, P<0. 05 vs control group. DEX significantly increased the expression of Foxp3 mRNA (P<0. 05) compared with AR and control group.@*CONCLUSION@#DEX reduce upper airway allergic inflammation effectively, which may be mediated by promoting the expression of Foxp3 and inducing the amplification of Tregs in vivo.


Subject(s)
Animals , Mice , Administration, Intranasal , Dexamethasone , Pharmacology , Disease Models, Animal , Flow Cytometry , Forkhead Transcription Factors , Metabolism , Inflammation , Drug Therapy , Mice, Inbred BALB C , Nasal Mucosa , Ovalbumin , RNA, Messenger , Rhinitis, Allergic , Drug Therapy , T-Lymphocytes, Regulatory
6.
Journal of Clinical Otorhinolaryngology Head and Neck Surgery ; (24): 1134-1136, 2015.
Article in Chinese | WPRIM | ID: wpr-747250

ABSTRACT

Metastatic tumors to the nasal cavity and paranasal sinuses are far less common than primary cancer in this location. The digestive system malignant tumor metastasis to the nose is rarer. The clinical presentation of metastases is similar to that of primary tumors and common symptoms include recurrent epistaxis, nasal obstruction and facial pain. Metastases to the nose and paranasal sinuses usually respond poorly to treatment and have a poor prognosis.


Subject(s)
Humans , Carcinoma , Pathology , Digestive System , Nasal Cavity , Pathology , Nasal Obstruction , Nose Neoplasms , Paranasal Sinus Neoplasms , Paranasal Sinuses , Pathology , Stomach Neoplasms , Pathology
7.
Chinese Journal of Primary Medicine and Pharmacy ; (12): 1169-1170, 2010.
Article in Chinese | WPRIM | ID: wpr-389295

ABSTRACT

Objective To investigate clinical efficacy of the Kirschner wire and nickel-titanium tension band fixation of patellar different types of patellar fracture.Methods 22 cases of patellar fractures were followed up,analyzed.Two methods of clinical treatment were compared.Results There was no significant difference between two methods of treatment.Transverse patella fractures with Kirschner wire tension band fixation was satisfied,but comminuted fracture of patella should adopt the nickel-titanium poly patella fixation.Conclusion Different types of patella fractures should be based on surgical indication to select different method of internal fixation.

8.
Chinese Journal of Anesthesiology ; (12): 461-464, 2010.
Article in Chinese | WPRIM | ID: wpr-388837

ABSTRACT

Objective To investigate the role of Notch signaling pathway in cerebral ischemic tolenmce induced by clectroacupuncture (EA) preconditioning.Methods Fifty-two adult male SD rats weighing 280-320 g were randomly divided into 2 groups(n=each):control group(group C)and electroacupuncture preconditionig group(group EA).Group C received no treatment.Group EA received EA at the Baihui acupoint (GV20) for 30 min a day for 5 days.Twenty-four hours after the last preconditionig,focal cerebral ischemia was induced by middle cerebral artery occlusion (MCAO) for 120 min,followed by 72 h of reperfusion.Notch intracellular domain(NICD)expression was determined by Western blot and expression of Notch1,Notch4,Jag1,and Hes1 mRNA by real-time PCR immediately before iachemia and 24 and 72 h of reperfusion.The neurological deficit was scored at 72 h of reperfusion.The infarct volumes were then determined after evaluation of the neurological deficit score .Results There was no significant difference in Hesl mRNA and NICD expression immediately before ischemia between group EA and C(P>O.05).NICD expression was up-regulated at 24 and 72 h of reperfusion in both groups, and Hesl mRNA expression at 72 h of reperfusion in group C and at 24 h of reperfusion in group EA was up-regulated compared with those immediately before ischemia (P < 0.05). Hes1 mRNA and NICD expression was up-regulated in group C, while down-regulated in group EA at 72 h of reperfusion compared with those at 24 h of reperfusion ( P < 0.05 ). Compared with group C, the expression of Notchl,Notch4 and Jag1 mRNA was up-regulated immediately before ischemia, and Hes1 mRNA and NICD expression was up-regulated at 24 h of reperfusion while down-regulated at 72 h of reperfusion in group EA( P < 0.05). EA preconditioning significantly reduced infarct volumes and increased neurological deficit scores at 72 h of reperfusion (P < 0.05 ). Conclusion Notch signaling pathway may participate in cerebral ischemic tolerance induced by EA preconditioning.

9.
Chinese Journal of Anesthesiology ; (12): 473-476, 2010.
Article in Chinese | WPRIM | ID: wpr-388835

ABSTRACT

Objective To investigate the effect of methylene blue(MB)on liver ischemia-reperfusion (I/R) injury in rabbits.Methods Twenty-four healthy New Zealand white rabbits of both sexes welshins 2.0-2.3 kg were randomly divided into 3 groups (n=8 each):group Ⅰ sham operation(group s);group Ⅱ I/R and group Ⅲ methylene blue (group MB).The animals were anesthetized with intravenous 2% pentobarbital 30 mg/kg.Liver I/R was produced by occlusion of hepatic blood flow for 40 min followed by 60 min repeffusion.In group MB methylene blue 5 mg/kg was injected iv at 20 min before liver ischemia.Femoral artery was carmulated for MAP monitoring and blood sampling.MAP and HR were recorded immediately before(T1,baseline)and at 20 and 40 min of ischemia (T2,3) and 1,5,30,60 min(T4-7)of repedusion.Blood samples were collected at T1,T5,T6 and T7 for measurement of seruln TNF-α and IL-6 concentrations.Plasma AST and ALT activities were measured at T1,T6 and T7.Liver specimens were obtained at the end of experiment for determination of SOD activity and MDA content.Results In group I/R MAP was significantly decreased at T4-7 during reperfusion and HR at T7 as compared with the baseline at T1;while in group MB no significant change in MAP and HR Was observed during ischemia and reperfusion as compared with the baseline.The gerum TNF-α and IL-6 concentrations and the plasma ALT and AST activities were significantly increased during reperfusion as compared with the baseline immediately before ischemia in group I/R and MB and were significantly lower in group MB than in group. I/R. The SOD activity was significantly higher while MDA content was significantly lower in group MB than in group I/R. Microscopic examination showed that liver damage was less severe in group MB than in group I/R. Conclusion The administration of MB can maintain hemodynamic stability and attenuate liver I/R injury in rabbits.

10.
Chinese Journal of Orthopaedics ; (12): 658-661, 2009.
Article in Chinese | WPRIM | ID: wpr-394108

ABSTRACT

Objective To discuss the characteristics, treatment method and prognosis of tibial tuberosity avulsion fracture in children. Methods From January 1995 to December 2007, there were 25 cases (23 boys and 2 girls). The age at injury ranged from 12 to 16 years(average 13.5 years). All cases were unilateral involved (10 of left, 15 of right). All of the injuries resulted from participation in athletic activities, 10 in basketball, 7 while jumping, and 8 in football game. According to the classification of Ogden, there were 1 type 1A, 4 type 1B, 5 type 2A, 7 type 2B, 4 type 3A, and 4 type 3B fractures. One type 1A fracture was treated nonoperatively with east immobilization, and other 24 fractures with open reduction and internal fixation. Results All of 25 cases were followed from 14 months to 7 years (average 43 months). The range of motion of knee joint was 0°-140° 3 months after operation, and all of them recovered to pre-injury levels of activity by 6 months after operation. According to Mosier clinical assessment system, the final outcome was evaluated as excellent in all. Wound healed primary, no compartment syndrome was found. The time of bony union ranged from 2 to 5 months, with the mean time of 3 months. No complications of infection, ex-tremities deformity and inner fixation failure were found. Conclusion The tuberosity avulsion fracture in children is an uncommon and special type of epiphyseal injury. These fractures occur in the narrow range of 13 to 16 years of age. The majority of patients are males, and most of them are unilateral involved. This in-jury has an excellent outcome without deformity or functional loss, once adequate reduction and fixation of fracture fragments have been achieved.

11.
Journal of Clinical Otorhinolaryngology Head and Neck Surgery ; (24): 488-490, 2009.
Article in Chinese | WPRIM | ID: wpr-748698

ABSTRACT

OBJECTIVE@#To investigate the efficacy of revision endoscopic sinus surgery and combined therapy on recurrent sinusitis and polyps.@*METHOD@#Revision endoscopic sinus surgery was performed in 72 patients, of which endoscopic nasal lateral wall dissection was used in 3 cases, the endoscopic frontal sinus surgery (Draf I-II) was used in 16 cases, and all patients received combined therapy including peri-operation conservative management and nasal endoscopy examination during the follow-up period.@*RESULT@#All patients were followed up for more than one year. Of 72 patients, 52 patients were successfully cured, 10 patients showed improvement, but there was no change in other 13 patients. The total efficacy rate was 91.67% (66/72). No serious complication occurred.@*CONCLUSION@#The treatment efficacy can be greatly improved by enough preoperative preparation, fine operation, combined pre-operation conservative therapy and postoperative follow-up.


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Endoscopy , Nasal Polyps , General Surgery , Recurrence , Sinusitis , General Surgery
12.
Chinese Journal of Anesthesiology ; (12): 1036-1039, 2009.
Article in Chinese | WPRIM | ID: wpr-391578

ABSTRACT

Objective To investigate the effect of different programs of preconditioning with hyperbaric oxygen (HBO) on spinal cord ischemia-reperfusion injury (I/R) in rabbits. Methods Forty-five New Zealand rabbits aged 4-5 months weighing 2.0-2.5 kg were randomly divided into 5 groups: group S, sham operation ( n = 5);group IR, spinal cord I/R injury (n = 10);group H_(1~3) , the animals were pretreated with 100% O_2 at 2.5 ATA 1 h/d for 5 (group H_1 ), 10 (group H_2 ) , or 20 (group H_3 ) consecutive days respectively 24 h before spinal cord I/R. The animals were anesthetized with iv pentobarbital sodium 30 mg/kg. The artery in the ear and left femoral artery were cannulated for proximal and distal mean blood pressure monitoring. Spinal cord ischemia was produced by cross-clamping of abdominal aorta distal to renal artery for 20 min. Hind-limb motor function was assessed at 48 h after reperfusion according to the modified criteria established by Tarlov (0 = no spontaneous movement, 4= normal motor function) . The animals were then killed and the L_5 segment of the spinal cord was removed for detection of neuronal survival (by HE staining), apoptosis (by TUNEL) and degeneration (by Fluoro-Jade B staining). Results Preconditioning with 5 or 10 d of HBO improved the hind-limb motor function and preserved more normal neurons in the spinal cord after I/R injury. Both apoptotic and degenerative cell death were attenuated in H_1 and H_2 groups. There was no significant difference in hind-limb motor dysfunction and the number of normal neurons in the lumbar spinal cord between H_3 group and I/R group. Conclusion Preconditioning with 5 d or 10 d HBO induces tolerance against spinal cord I/R injury, whereas preconditioning with 20 d of HBO fails to protect the spinal cord from I/R injury.

13.
Chinese Journal of Medical Education Research ; (12)2006.
Article in Chinese | WPRIM | ID: wpr-623559

ABSTRACT

The standardized training for residents is the key point of medical education and clinical level enhancement.The future clinicians not only must have the solid specialized knowledge,but also should have the rich humanities and the social sciences knowledge,good interpersonal communication ability,as well as lofty occupational ethics.Therefore,how to train residents to be rich in both the spirit of science and humanity and the knowledge of science and humanity,has become an important topic in the clinical medicine education.

14.
Chinese Medical Journal ; (24): 108-111, 2003.
Article in English | WPRIM | ID: wpr-356858

ABSTRACT

<p><b>OBJECTIVE</b>To investigate whether pretreatment with repeated electroacupuncture (EA) at the Baihui acupoint could induce ischemic tolerance against transient focal cerebral ischemic injury in rats.</p><p><b>METHODS</b>Thirty male Sprague-Dawley (SD) rats were randomly divided into 3 groups (n=10 for each): the control group consisted of animals receiving no treatment, the isoflurane (ISO) group had animals that inhaled 1.5% isoflurane for 30 min a day for 5 days, and animals in the EA group received electroacupuncture at the Baihui acupoint for 30 min a day for 5 days under 1.5% isoflurane anesthesia. Twenty-four hours after the last treatment, the middle cerebral artery was occluded with No. 3 nylon monofilament for 120 min. The neurological outcomes were evaluated 24 h after reperfusion. The infarct volumes were then assessed using 2% triphenyltetrazolium chloride staining after the neurological outcome evaluation.</p><p><b>RESULTS</b>The neurological deficit score (NDS) of the EA group [1 (0-2)] was lower than that of the ISO group [2 (1-3)] and the control group [2 (1-4)], P < 0.05. The infarct volume of the EA group (38.3 +/- 25.4 mm(3)) was significantly smaller than that of the control group (220.5 +/- 66.0 mm(3)) and the ISO group (168.6 +/- 57.6 mm(3)) 24 h after reperfusion.</p><p><b>CONCLUSION</b>Electroacupuncture at the Baihui acupoint 30 min a day for 5 days significantly reduces neurological injury induced by transient middle cerebral artery occlusion.</p>


Subject(s)
Animals , Male , Rats , Electroacupuncture , Ischemic Attack, Transient , Therapeutics , Rats, Sprague-Dawley
15.
Journal of Audiology and Speech Pathology ; (6)1998.
Article in Chinese | WPRIM | ID: wpr-533993

ABSTRACT

Objective To analyze the results in neonatal hearing screening,and then to make diagnosis and intervention for neonates with hearing problems as soon as possible.Methods From January 2004 to December 2008,7 064 newborns at Shanghai East Hospital received hearing screening by distortion product oto-acoustic emissions (DPOAE) 2~4 days after birth.Re-screening tests were performed for infants who failed the initial screening 42 days after birth.Those newborns who failed again received further audiologic diagnostic evaluations 3 months afterwards.Results Among 7 064 cases screened,out of 6 412 normal newborns,579 (9.03%)failed the initial screening,and 38 (7.41%)failed re-screening.129 cases of the other 652 (19.79%) newborns in NICU failed the initial screening,and 20 cases (18.69%) failed re-screening.There were statistical differences between the passing rates of the two groups (P

16.
Chinese Journal of Anesthesiology ; (12)1997.
Article in Chinese | WPRIM | ID: wpr-517244

ABSTRACT

Objective To investigate the dose response effects of Shenfu injection, an intravenous drug made from traditional Chinese herbs, on the ischemic spinal cord injury in rabbits Methods New Zealand white rabbits were anesthetized with halothane and spinal cord ischemia was induced with 20 min infrarenal aortic occlusion Animals were randomly allocated to 4 groups, in group A(n=6) without pharmacologic intervention, in group B (n=6) ,group C (n=6) and group D (n=6) Shenfu injection 5,10 and 20ml?kg -1 were infused intravenously at a constant rate within 30min before the aortic cross clamping, respectively Neurologic status was scored by the Tarlov system 1,4,8,12,24 and 48h following reperfusion The animals were sacrificed 48h following reperfusion to sample the spinal cord (L5 7) immediately for histopathologic study Results All animals survived during the experiment Compared with that in group A, the neurologic deficit score (NDS) increased significantly at each observing time in group B, group C and group D (P0 05), but there was no significant difference between group B, C and D NDS in group A,B,C and D 48h after reperfusion were 0 5?0 8,3 2?0 9(P

17.
Chinese Journal of Anesthesiology ; (12)1995.
Article in Chinese | WPRIM | ID: wpr-517306

ABSTRACT

Objective To investigate if isoflurane preconditioning induces ischemic tolerance against neuronal damage produced by middle cerebral artery occlusion (MCAO) in rats Methods Thirty male SD rats weighing 350 400g were randomly divided into three groups: control group (no pretreatment,n=10),isoflurane preconditioning group (ISO group) (inhalation of 2% isoflurane and 98% O 2 1h per day lasting 5d,n=10), oxygen preconditioning group (O 2 group) (inhalation of 98% O 2 1h per day lasting 5d ,n=10) Right MCAO was induced by a 3 0 nylon thread with round tip inserted cranially into right internal carotid artery under isoflurane anesthesia and maintained for 120 min The neurologic deficit score (NDS) was evaluated 1, 3, 6, 12, 16 and 24h after reperfusion and the infarct volume was calculated at 24th following reperfusion Results The NDS of ISO group was lower than that of other two groups at each time interval (P0 05) Conclusions Isoflurane pretreatment can induce ischemic tolerance against neuronal damage produced by transient MCAO in rats

18.
Acta Anatomica Sinica ; (6)1957.
Article in Chinese | WPRIM | ID: wpr-575606

ABSTRACT

Objective To investigate the effect of stress induced by water restriction on the level of phosphorylated(ERK1/2) in rat brain. Methods Fifty-eight male Wistar rats were used and randomly divided into control group(C,n=4) and three experimental groups: TW(drinking at scheduled time),EB(empty bottle stimulus),WR(restricted water drinking),which were subdivided into three subgroups by time as 3d,7d, 14d(n=6 for each).In C group,water was freely accessed to the animal for one week.In the experimental groups the water supply was restricted following a week of accommodation and the other week for timed water supply training.In TW group,the water was supplied twice a day and 10 min for each in a fixed time. In EB and WR groups,the water was served only once a day for 10 min,and in the other scheduled time for water supply only an empty bottle without water was provided in the EB group and water was limited in the WR group.All the animals were deeply anesthetized 1h after the last stimulus and transcardially infused,then the brains were cut into free-floating sections and processed for immunohistochemical staining of pERKI/2 with ABC method.The expression of pERK1/2 positive staining was measured with image analysis system. Results The area of phosphorylated ERK1/2 was significantly increased in many brain structures,especially in paraventricular hypothalamic nucleus(PVN),supraoptic nucleus(SON),ventrolateral septum(LSV),medial amygdaloid nucleus(MeA),central amygdaloid nucleus(CEA) and nucleus of solitary tract(NTS) in TW,EB and WR group compared with C group.The expression of phosphorylated ERK1/2 in SON,LSV, MeA and CeA in EB and WR group were higher than that in TW group;pERK1/2 expression of LSV,MeA,CeA and NTS in EB group increased and went down more early and quickly than that in WR group.The response of phosphorylated ERK1/2 was dependent on both stimulus methods and time in all the nuclei mentioned above except PVN,in which only the stimulus time was related. Conclusion The level of pERK 1/2 was significantly altered in PVN,SON,LSV,MeA,CeA and NTS following stimulation of water restriction.The results indicate that the activated nuclei in the brain by physical and psychological stressors are different.

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