To investigate the effects of substance P in globus pallidus on haloperidol-induced Parkinson's disease model rat.Methods In behavioral experiments,guide cannulae constructed from stainless steel were implanted into the globus pallidus.After five days recovery,0.5 μl drugs(normal saline,SMSP,SR140333B,SR140333B + SMSP)were bilaterally microinjected into the globus pallidus in awake rats with haloperidol administration intraperitoneally.The catalepsy was then observed within 60 min.In electrophysiological study,an in vivo extracellular recording was performed to observe the effects of substance P on the firing rate of pallidal neurons.Resuits Haloperidol induced catalepsy in rats with intrapallidal saline microinjection.The maximum average latency was(259.8±34.8)s at the time point of 30th min.The minimum average latency was(145.2±54.8)s at 50th min.Bilateral microinjection of SMSP into globus pallidus significantly attenuated haloperidol-induced catalepsy (The average latency was(10.4±3.4)s at lOth rain and(58.4±38.8)s at 60th min,P＜0.01).This anticataleptic effect was completely counteracted by selective NK-1 receptors antagonist SR140333B(The average latency was(176.4±64.4)s at 10th min and(139.2±59.7)s at 60th rain,P＜0.01).Furthermore,micropressure ejection of SMSP significantly increased the firing rate of pallidal neurons(Basal:(13.4±4.2)Hz,SMSP:(17.5±5.6)Hz).The average increase was(29.4±8.6)%(P＜0.05,n=13).SR140333 B completely blocked SMSPinduced increase in firing rate(SR140333B:(10.3±2.5)Hz,SR140333 B + SMSP:(11.3±3.0)Hz,P＞0.05,n=8).Conclusion Based on the action of substance P in globus pallidus of parkinsonian rats,it is we hypothesized that activation of substance P receptor in globus pallidus may play a role in the treatment of Parkinson's disease.

Objective:To analyze the clincial characteristics and laboratory findings of patients with neurobrucellosis (NB).Methods:Using retrospective analysis, clinical diagnosed patients with NB from June 2016 to February 2019 in Heilongjiang Agricultural Reclamation Bureau General Hospital were selected to analyze the general characteristics, clinical symptoms, laboratory examination results [white blood cell (WBC), hemoglobin(Hb), c-reactive protein (CRP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total protein (TP), albumin (ALB), cerebrospinal fluid routine and biochemical, serum tube agglutination test (SAT), blood culture and cerebrospinal fluid culture of Brucella] , diagnosis and treatment effect. Results:A total of 25 patients were diagnosed with NB, including 19 males and 6 females, with an average age of (41.7 ± 14.2) years old, ranged from 11 to 70 years old. The main clinical symptoms were fever, headache, joint pain, vomiting and sweating, which accounted for 92.0% (23/25), 88.0% (22/25), 76.0% (19/25), 64.0% (16/25), and 64.0% (16/25), respectively. Positive neck ankylosis and mumbness of lowerlimbs were both 9 cases (36.0%), and mental disorders were 7 cases (28.0%). In 25 patients with NB, the WBC increased in 5 cases (20.0%), Hb decreased in 4 cases (16.0%), CRP increased in 13 cases (52.0%), ALT and AST both increased in 6 cases (24.0%), TP decreased in 21 cases (84.0%); SAT was positive in 25 cases (100.0%), cerebrospinal fluid SAT positive in 7 cases (28.0%); and blood culture was positive in 2 cases (8.0%). Cerebrospinal fluid changes were mainly manifested in 14 cases (56.0%) of chloride reduction, 13 cases (52.0%) of gluose reduction and 19 cases (76.0%) of protein increase. In 25 patients with NB, 17 cases were treated with doxycycline + rifampicin + ceftriaxone, 7 cases with etimicin + rifampicin + ceftriaxone, and 1 case with doxycycline + rifampicin + piperacillin sulbactam. After 6 to 12 months follow-up, 21 cases recovered well, whereas mild sequelae were observed in 4 patients.Conclusion:Clinical features of NB are hetorogeneous, and nerurological symptoms and cerebrospinal fluid examination are of great value in the diagnosis of NB.

Objective To investigate the effect of acute myocardium ischemic on heart function of pregnancy rat.Methods 13 female SD rats and 6 early pregnancy rats were divided into normal group, unpregnant group with acute myocardial infarction and early pregnant group with acute myocardial infarction. The anterior branch of the left coronary artery was ligated. 3 weeks later, Image 1.31 software was used to measure areas of myocardial infarction,and to evaluate hemodynamics of heart with powerLAB4.12, and cardiac tissues were stained with Massion. Results Compared with unpregnant group with acute myocardial infarction , the early pregnant group with acute myocardial infarction had less myocardial infarction area (28. 86% vs. 36. 8%), and had a higher left ventricle end systolic pressure, ±dp/dt max, and lower left ventricle end diastolic pressure. Massion stain showed the amount of collagen of the lesion was less in the early pregnant group with acute myocardial infarction than that in unpregnant group.Conclusion The early pregnant group with acute myocardial infarction had better heart contractive and diastolic function.

Objective To analyze the epidemiology and clinical characteristics of Brucella infection in Heilongjiang Province,and to provide experience and basis for clinical treatment and prevention of brucellosis.Methods Using retrospective analysis method,clinically diagnosed patients with brucellosis from December 2016 to December 2017 in Heilongjiang Agricultural Reclamation Bureau General Hospital were selected to analysis the general characteristics,clinical symptoms,laboratory examination results,diagnosis and treatment effect.Healthy subjects undergoing health check-up at the same period were randomly selected as control group for laboratory blood biochemical test.Results A total of 396 patients were included in the brucellosis group,including 298 males and 98 females,with an average age of (43.6 + 15.6) years old,ranged from 1 to 75 years old.The control group included 125 cases,including 83 males and 42 females,with an average age of (41.7 ±+ 18.0) years old,ranged from 7 to 71 years old.There were no significant differences in age and gender between the two groups (t =1.083,x2 =3.789,P ＞ 0.05).The patients of ＞ 14-64 years old of age had the highest incidence (90.15％,357/396).The patients were mainly rural residents (76.01％,301/396) and the majority of them were farms (50.51％,200/396).The onset time was mainly from March to June of the year (47.73％,189/396).Clinical symptoms were fever,joint or muscle pain and hyperhidrosis,which accounted for 91.16％ (361/396),76.26％ (302/396) and 44.19％ (175/396),respectively.The levels of C-reactive protein (CRP),seropropane alanine aminotransferase (ALT),aspartate transaminase (AST),γ-glutamyl transpeptadase (γ-GT),alkaline phosphatase (ALP),α-hydroxybutyrate dehydrogenase (HBDH),and lactate dehydrogenase (LDH) in patients with brucellosis were significantly higher than those in the control group (t =4.956,2.746,4.019,3.272,7.008,3.728,3.409,P ＜ 0.01 or 0.05),while total protein (TP) and albumin (ALB) levels were reduced (t =-7.252,-8.601,P ＜ 0.01).After treatment,the test results of CRP,ALT,AST,γ-GT,TP,ALB,ALP,HBDH and LDH were compared with those before the treatment,and the differences were statistically significant (t =2.291,4.393,3.382,2.608,-7.516,-4.710,3.509,4.585,4.473,P ＜ 0.01 or 0.05).Conclusions The patients with brucellosis in Heilongjiang Province are mainly male farmers,and the onset time is mostly concentrated in March-June.The clinical manifestations of brucellosis are varied,clinicians should strengthen discrimination and prevent misdiagnosis.Medicines for brucellosis might be cautiously used in clinical trial,and changes in liver function and myocardial enzyme activity should be monitored.

Objective: To analyze the epidemiologic and clinical features of preschool children with brucellosis, and improve awareness of brucellosis and level of diagnosis. Methods: In-patients treated for brucellosis in preschool children from December 2016 to November 2018 in Heilongjiang Agricultural Reclamation Bureau General Hospital were retrospectively analyzed for epidemiology, clinical feature, laboratory data, treatment and prognosis. Results: There were 29 males and 16 females among the 45 preschool children brucellosis cases, with an average age of (3.7 ± 1.6) years old, ranged from 6 months to 6 years old. The patients of > 3-6 years old of age had the highest incidence (64.44%, 29/45). The patients were mainly rural residents (97.78%, 44/45). The onset time was mainly from March to June of the year (46.67%, 21/45). Clinical symptoms were mostly fever and joint pain, which accounted for 97.78% (44/45) and 57.78% (26/45), respectively. And lymph node enlargement and hepatosplenomegaly were also commonly seen, which accounted for 42.22% (19/45) and 35.56% (16/45). After treatment, the test results of patients with alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatine kinase isoenzyme (CK-MB), α-hydroxybutyrate dehydrogenase (HBDH) and lactate dehydrogenase (LDH) were compared with those of before the treatment, the differences were statistically significant (t=4.774, 2.970, 2.229, 5.664, 5.805, P < 0.05). Before the treatment, 36 patients (80.00%, 36/45) had positive blood culture, which turned negative after the treatment. Conclusions The epidemiologic and clinical features of preschool children with brucellosis are not typical. In the epidemic area of the disease and multiple seasons, for children with unexplained fever, the clinician should inquire about the epidemiological history and combine the clinical feature, consider the possibility of brucellosis, and strive for early diagnosis and treatment.

Objective To prepare a rabbit anti-mouse coiled-coil domain containing 189 (Ccdc189) polyclonal antibody. Methods The pET-28a-Ccdc189 prokaryotic expression plasmid was constructed and transformed into E.coli BL21. IPTG was used to induce the expression of Ccdc189 prokaryotic protein. Adult male New Zealand rabbits were immunized with purified recombinant protein to obtain rabbit anti-mouse Ccdc189 polyclonal antibody. The specificity of the polyclonal antibody was identified by Western blot analysis, indirect ELISA and immunofluorescence histochemical staining. Results The pET-28a-Ccdc189 recombinant plasmid was successfully constructed and the expression of the Ccdc189 recombinant protein was induced. ELISA revealed that the titer of the polyclonal antibody was 1:1 000 000. Western blot and immunofluorescence staining demonstrated that the Ccdc189 polyclonal antibody could specifically identify the Ccdc189 prokaryotic protein and the Ccdc189 protein in adult wild-type mouse testis. Conclusion A polyclonal antibody with high specificity against mouse Ccdc189 was successfully created.
Rabbits ; Male ; Animals ; Mice ; Antibody Specificity ; Antibodies ; Enzyme-Linked Immunosorbent Assay ; Blotting, Western ; Recombinant Proteins ; Escherichia coli/genetics*

Objective To produce rabbit polyclonal antibody against mouse testis expressed 38 (TEX38). Methods Full-length open reading frame sequence of TEX38 was amplified and inserted into the pET-30a-(+) vector to construct pET-30a-TEX38 prokaryotic plasmid. The recombinant plasmid was transformed into E.coli BL21, and expression was induced with isopropyl β-D-thiogalactopyranoside (IPTG). New Zealand white rabbits were immunized with TEX38 protein after purification and denaturation, then TEX38 polyclonal antibodies were collected from rabbit serum samples. ELISA was performed to detect the antibody titer. Western blot and immunofluorescence staining were performed to determine the specificity of TEX38 polyclonal antibodies. Results The pET-30a-TEX38 recombinant plasmid was constructed, and TEX38 prokaryotic protein was expressed and purified successfully. After immunization, the titer of TEX38 antibody reached 1:1 000 000. Western blot analysis and immunofluorescence staining showed that TEX38 was localized in the mouse spermatogenic cells and sperms with a good specificity. Conclusion The rabbit polyclonal antibody against mouse TEX38 is successfully produced, and the expression of TEX38 in mouse spermatogenic cells and sperms is validated.
Male ; Rabbits ; Animals ; Mice ; Testis ; Antibodies ; Enzyme-Linked Immunosorbent Assay ; Immunization ; Spermatozoa ; Escherichia coli

Objective To prepare rabbit polyclonal antibody against mouse IQ and ubiquitin-like domain-containing protein (IQUB) and detect its expression in the mouse testis. Methods Full-length coding sequence of IQUB was inserted into the pET-30a(+) vector to construct pET-30a-IQUB recombinant prokaryotic plasmid. Transformation of pET-30a-IQUB plasmid into E. coli BL21 was performed, and protein expression was induced with isopropyl-beta-D-thiogalactoside (IPTG). The protein was purified through histidine-tagged fusion protein purification column, then denatured by treatment of urea with gradient concentration. New Zealand rabbits were immunized with the denatured protein to produce IQUB polyclonal antibody. Antibody titer was detected by ELISA, and Western blot analysis and immunofluorescence assay were employed to validate the effectiveness and specificity of IQUB antibody. Results pET-30a-IQUB recombinant plasmid was constructed, and protein expression of IQUB was induced successfully with IPTG. The titer of IQUB polyclonal antibody reached 1:1 000 000. The antibody specifically recognized the endogenous IQUB protein of testis in the wild-type adult mouse. IQUB was expressed in spermatogenic cells of different stages. It was localized in the acrosome and flagellum of mature sperms. Conclusion The highly specific rabbit anti-mouse IQUB polyclonal antibody is successfully prepared, which can be used for Western blot and immunofluorescence histochemistry.
Male ; Rabbits ; Animals ; Mice ; Ubiquitins ; Escherichia coli/genetics* ; Isopropyl Thiogalactoside ; Antibodies ; Enzyme-Linked Immunosorbent Assay