Germline PRKACA amplification-associated primary pigmented nodular adrenocortical disease: a case report and literature review

SUMMARY Primary pigmented nodular adrenocortical disease (PPNAD) is a rare adrenocorticotropin hormone (ACTH)-independent Cushing's syndrome (CS). Pediatric patients with PPNAD typically have unusual skin lesions and slow growth with unknown causes. We present a case of a female Chinese patient with PPNAD caused by the germline PRKACA gene copy number gain of chromosome 19. The patient initially presented with kidney stones, short stature, and obesity. After further testing, it was discovered that the patient had diabetes, mild hypertension, low bone mass, a low ACTH level, and hypercortisolemia, and neither the low-dose or high-dose dexamethasone suppression test was able to inhibit hematuric cortisol, which paradoxically increased. PPNAD was pathologically diagnosed after unilateral adrenalectomy. Chromosome microarrays and whole exon sequencing analyses of the peripheral blood, as well as testing of sectioned adrenal tissue, showed a rise in the copy number of the duplication-containing PRKACA gene on chromosome 19p13.13p13.12, a de novo but not heritable gene defect that causes disease. The clinical signs and symptoms supported the diagnosis of Carney complex (CNC). One significant mechanism of CNC pathogenesis may be the rise in germline PRKACA copy number of chromosome 19. When assessing PPNAD patients for CNC, the possibility of PRKACA gene amplification should be considered. The effect of PRKACA gene amplification on the clinical manifestations of CNC needs to be confirmed by more cases.

Resveratrol improves cognitive function in severely burned rats by inhibiting hippocampal NF-κB/JNK pathway / 细胞与分子免疫学杂志

Objective To investigate the protective effect of resveratrol (RSV) on improving cognitive function in severely burned rats and its possible mechanism. Methods 18 male SD rats aged 18-20 months were randomly divided into 3 groups: control group, model group and RSV group, with 6 rats in each group. After successful modeling, the rats in RSV group were gavaged once daily with RSV (20 mg/kg). Meanwhile, the rats in control group and model group were gavaged once daily with an equal volume of sodium chloride solution. After 4 weeks, the cognitive function of all rats was estimated by Step-down Test. The concentration of tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) protein in serum of rats were detected by ELISA. The expression of IL-6, TNF-α mRNA and protein were estimated by real-time PCR and Western blotting. The apoptosis of hippocampal neurons was tested by terminal deoxynuclectidyl transferase-mediated dUTP-biotin nick end labeling assay (TUNEL). The expression of nuclear transcription factor-κB (NF-κB)/c-Jun N-terminal kinase (JNK) pathway-related proteins in hippocampus were assessed by Western blotting. Results Compared with the rats in model group, rats in RSV group exhibited improved cognitive function. Consistently, the rats in RSV group had a reduced concentration of TNF-α and IL-6 in serum, decreased mRNA and protein expressions of TNF-α and IL-6 in hippocampus, and decreased apoptosis rate and relative expression of p-NF-κB p65/NF-κB p65 and p-JNK/JNK in hippocampal neurons. Conclusion RSV alleviates inflammatory response and hippocampal neuronal apoptosis by inhibiting NF-κB/JNK pathway, thereby improving cognitive function in severely burned rats.

Clinical effect of Traditional Chinese Medicine combined with pressure therapy in the treatment of eczema / 国际中医中药杂志

Objective:To observe the clinical effect of Traditional Chinese Medicine (TCM) collapsibility combined with pressure therapy in the treatment of eczema.Methods:From January 2021 to August 2021, 60 patients with lower extremity venous ulcers who were treated in Wangjing Hospital, China Academy of Chinese Medical Sciences were selected and randomly divided into the control group (30 patients) and observation group (30 patients). The control group was treated by the routine pressure therapy, and the observation group was by the TCM nursing intervention on the basis of the control group. Both groups were treated for 2 weeks. The visual analogy scale (VAS),chronic Venous Insufficiency Questionnaire (CIVIQ), Self-rating Anxiety Acale (SAS), and Self-rating Depression Scale (SDS) were measured and evaluated at baseline and after treatment.Results:The total response rate was 100.0% (30/30) in the observation group and 86.7% (26/30) in the control group, and the difference was statistically significant ( χ2=4.29, P=0.038). The VAS (2.47±0.68 vs. 2.97±0.14, t=3.95), SAS (40.31±11.93 vs. 55.87±4.53, t=4.73), SDS (44.30±5.38 vs. 47.61±5.33, t=2.39) scores were significantly lower than those in the control group ( P<0.05), CIVIQ score (86.63±2.74 vs. 84.67±3.23, t=2.54) was significantly higher than that of the control group ( P<0.05). Conclusion:TCM combined with pressure therapy can promote the healing of ulcers in patients with eczema, relieve pain, and improve the quality of life of patients.

Inhibitory effect of silencing HMGB1 gene on growth of human epithelial ovarian cancer xenografts in nude mice / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the effect of HMGB1 gene on the growth of human epithelial ovarian cancer xenografts in nude mice, and to lay a foundation for finding new targets for the treatment of ovarian cancer. Methods: Human epithelial ovarian cancer SKOV3 cells in logarithmic growth phase were selected to establish a human epithelial ovarian cancer xenograft model in nude mice. Nude mice with successful model establishment were randomly divided into control group and HMGB1-siRNA group. On the 7th, 9th, 11th, 14th, and 16th days after cell inoculation, the same amount of saline and HMGB1-siRNA were respectively injected into two groups of mice under the armpit.After 3 weeks, the nude mice were sacrificed by cervical dislocation, the tumor tissues were separated, and the volume of the tumor was measured. The apoptosis of transplanted tumor cells was detected by Tunnel staining. The expressions of HMGB1, STAT3 and p-STAT3 were detected by Western blotting. The expression of vascular endothelial growth factorA(VEGF-A) and microvascularization were detected by immunohistochemistry. Results: Compared with the control group, the growth of tumor volume slowed down in HMGB1 siRNA group, and on the 21st day, the tumor volume of HMGB1-siRNA group was significantly smaller than that of the control group (P<0.05). HMGB1-siRNA successfully knocked down the expression of HMGB1 mRNA in transplanted tumor tissue. The apoptosis rate of tissue cells in HMGB1-siRNA group was significantly increased ([34±8]% vs [6±2]%, P=0.04), and the expressions of HMGB1 and p-STAT3 were significantly reduced (P<0.05). The expression of VEGF-Aand the number of microvessels were significantly lower than those of the control group (both P<0.05). Conclusion: Knockdown of HMGB1 gene reduces the expression of VEGF-A and microvessel formation possibly by inhibiting the HMGB1/STAT3 signaling pathway, thereby promoting the apoptosis of tumor tissues and slowing the growth of xenografts.

Establishment of reference range for specific thyroid function during pregnancy and analysis of influencing factors of hypothyroxinemia / 中华内分泌代谢杂志

Objective To establish a reference range for specific thyroid function during pregnancy and to explore the influencing factors of hypothyroxinemia during pregnancy.Methods A retrospective analysis of 2 996 cases of thyroid function in the pregnant women who were with single pregnancy and without thyroid diseases and family history of those diseases.Results (1) Establish a unified reference range for specific thyroid function during pregnancy;the early,middle,and late trimesters thyrotropin (TSH) ranges were 0.02-6.39,0.16-6.23,0.64-6.59 mU/L,respectively,while free thyroxine (FT4) ranges were 11.32-23.00,9.39-18.92,8.54-16.73 pmol/L respectively.The specific reference ranges of Han and Uygur pregnant women were established separately.There was no difference in the detection rates of various thyroid diseases when using their respective reference ranges and the unified reference range of the hospital (P ＞ 0.05).(2) The detection rate of various thyroid diseases (except subclinical hyperthyroidism) of our subjects with China guideline reference range was significantly higher than the reference range with the hospital (P＜0.05).(3) The detection rates of hypothyroxinemia in all pregnant women with FT4 cut points of P2.5 and P5 were 4.3％ and 7.4％,respectively,of which the Han population was 4.3％ and 7.1％,respectively,and the Uygur population was 4.3％ and 7.9％,respectively.(4) Comparing the mean age,gestational age,median urine iodine,and thyroid antibody positive rate between the hypothyroxinemia group and the control group,only the mean age and gestational age were different (P＜0.05);Logistic binary regression analysis showed that age was the risk factor for hypothyroxinemia during pregnancy (OR =1.035,95％ CI 1.006-1.066,P ＜ 0.05).Conclusions The Han and Uygur pregnant women in this area both can use the thyroid reference range of our hospital during pregnancy.The establishment of thyroid reference range may avoid over-diagnosis of thyroid disease during pregnancy.Age is a possible influencing factor of hypothyroxinemia during pregnancy.

Effect of family education methods and parental rearing or not on the psychological problem tendency of primary school students / 中华行为医学与脑科学杂志

Objective To explore the impact of family education methods and parental rearing or not on the psychological problem tendency of primary school students.Methods The mental health test (MHT) was used on getting mental health data of the 2 838 primary school students from Dongguan city.Results (1) The results of MHT showed that 94.28％ of primary school students had no psychological problems,the tendency rate of psychological problems was 4.29％,and 1.43％ of them were suffering from serious psychological problems.Learning anxiety,physical symptoms and allergy tendency were more prominent.(2) The total MHT scores of primary school students with authoritarian,democratic,permissive and neglected family education methods were (34.84±14.58),(31.04±13.60),(35.19±12.82) and (41.19±13.10)respectively.There was no statistically significant difference in the terrorist tendency dimension scores of primary school students with different education methods (F=2.33,P=0.054),and the differences in the MHT total score and other dimensions were statistically significant (F=4.35-16.88,P＜0.01).(3) There were statistically significant differences in the scores of the total score of psychological problem tendency,dimensions of learning anxiety,anxiety to people,allergy tendency and impulse tendency of primary school students who were parental rearing or not (t=2.09-3.67,P＜0.05).However,there was no statistically significant difference in the dimensions of loneliness tendency,self-accusation tendency,physical symptoms and terrorist tendency (P＞0.05).Conclusion Family education methods and parenting styles has a impact on the mental health of primary school students.

Protective effect of salidroside on rats with hypoxic pulmonary hyperten-sion by suppressing oxidative stress / 中国病理生理杂志

AIM:To study whether salidroside plays a protective role in hypoxia-induced pulmonary hyperten-sion by suppressing oxidative stress.METHODS: Sprague-Dawley rats were randomly divided into 4 groups: normoxia (N)group,hypoxia for 4 weeks(H4)group,low-dose salidroside(hypoxia for 4 weeks and treatment with salidroside at 16 mg/kg,H4S16)group and high-dose salidroside(hypoxia for 4 weeks and treatment with salidroside at 32 mg/kg, H4S32)group.The mean pulmonary arterial pressure(mPAP), the weight ratio of right ventricle/(left ventricle+sep-tum)[RV/(LV+S)]and vessel wall area/vessel total area(WA/TA)were evaluated.The levels of malondialdehyde (MDA)in the serum and lung tissues were detected by colorimetric method.The levels of 8-iso-prostaglandin F2α(8-iso-PGF2α)in the serum and lung tissues were measured by ELISA.The activity of superoxide dismutase(SOD)in the serum was analyzed by hydroxylamine method.The expression of NAPDH oxidase 4(NOX4)and SOD1 in the lung tissues was determined by Western blot.RESULTS:Compared with N group,the levels of mPAP,RV/(LV+S)and WA/TA in H4 group were significantly increased,which were apparently attenuated by salidroside injection in a dose-dependent manner. Meanwhile,salidroside administration apparently decreased the levels of MDA and 8-iso-PGF2αin the serum and lung tis-sues,as well as the expression of NOX 4 in the lung tissues.Besides,compared with N group, the activity of SOD in the serum and the expression of SOD1 in the lung tissues in H4group were significantly decreased,while administration of sali-droside increased the activity of SOD in the serum and the expression of SOD 1 in the lung tissues in a dose-dependent man-ner.CONCLUSION:Salidroside protects the pulmonary vessels from remodeling and attenuates hypoxia -induced pulmo-nary hypertension by inhibiting oxidative stress.

Effects of resveratrol on TNF-α-induced injury and expression of MCP-1 in isolated rat pulmonary artery endothelial cells / 中国病理生理杂志

AIM:To investigate the possible mechanism of resveratrol(Res)on tumor necrosis factor-α (TNF-α)-induced monocyte chemoattractant protein-1(MCP-1)expression in primary rat pulmonary artery endothelial cells(RPAECs).METHODS: RPAECs were randomly divided into 4 groups: control group, solvent(1% DMSO) group,TNF-αgroup and Res group.Each group was divided into 1 h,4 h and 8 h subgroups(n=6 per time point).The TNF-α+C1142(a rodent chimeric mAb that neutralizes rat MCP-1)group was set up at the 8 h time point.At each time point,the protein and mRNA expression of MCP-1 was measured by Western blot and real-time PCR.RESULTS: Pre-treatment of the RPAECs with C1142 significantly down-regulated the expression of MCP-1(P<0.05).The protein and mRNA expression of MCP-1 was markedly increased in TNF-αgroup(P<0.05).Notably,incubation with Res down-re-gulated the protein and mRNA expression of MCP-1,which was significantly lower than that in TNF-αgroup(P<0.05). CONCLUSION:MCP-1 was involved in the process of TNF-α-induced injury of RPAECs.Res down-regulates the expres-sion of MCP-1 in RPAECs,thus attenuating cell injury.

The therapeutic effect of hydrogen-rich saline on the rheological behavior of leukocytes in mesentery capillary of rats with high-voltage electrical burn / 国际检验医学杂志

Objective To investigate the therapeutic effect of hydrogen-rich saline on the rheological behavior of leukocytes in mesentery capillary of rats with high-voltage electrical burn .Methods 180 rats were randomly divided into four groups :burn injury plus normal saline group ,burn injury plus hydrogen-rich saline group ,sham plus normal saline group ,and burn injury plus papaver-ine group .The rats were received saline ,hydrogen-rich saline ,saline ,papaverine at different time points after scald respectively .The changes of rheological behavior of leukocytes in mesentery capillary of rats before and after the injury were investigated .Results The rheological behavior of leukocytes in mesentery capillary of the control group were observed no significant change (P>0 .05) . In experimental group the rolling white blood cell count ,the number of leukocyte adhesion ,the length of contact of leukocyte-endo-thelial cell at each phase after injury were higher than those at 15 min before injury (P<0 .05);leukocyte rolling speed after injury is lower than that before injury (P<0 .05) .In treatment group and positive control group ,the rolling white blood cell count ,the number of leukocyte adhesion ,the length of contact of leukocyte-endothelial cell at each phase after injury were higher than those at 15 min before injury (P<0 .05) ,but compared with the experimental group ,the increase range was lower (P<0 .05) .leukocyte rolling speed after injury is lower than that before injury (P<0 .05) ,and compared with the experimental group ,the reduction was lower (P<0 .05) .Conclusion The hydrogen-rich brine can effectively reduce the changes of rheological behavior of leukocytes in mesentery capillary of rats caused by high-voltage electrical burn ,and have a protective effect on rat mesenteric .

The study of catheterization to measure ventricular pressure in mice through venous cannula in external jugular vein / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To study a feasible method of measuring right ventricular pressure by catheterization in mice.</p><p><b>METHODS</b>Measuring the right ventricular pressure and the pulmonary artery pressure by homemade PE pipe through venous cannula in external jugular vein, using catheterization in mice with powerlab multimodal biometric signal recording system.</p><p><b>RESULTS</b>Forty-six out of 51 mice were experimented with this method smoothly and got a total success rate of 90.2%. Thirty of 33 normal mice and 16 of 18 mice with pulmonary arterial hypertension (PAH) were catheterized successfully. The right ventricular pressure were as follow: systolic blood pressure: (23.4 +/- 5.7) mmHg in normal group vs (32.2 +/- 2.8) mmHg in mice with PAH, diastolic blood pressure: (3.7 +/- 2.6) mmHg vs (3.8 +/- 2.0) mmHg, mean pressure: (12.0 +/- 3.7) mmHg vs (14.9 +/- 2.3) mmHg. After autopsy for those 5 failed cases, we found that 2 cases were into the inferior vena cava, another 2 cases pierced the right auricle and the last one punctured the axillary vein into the chest wall.</p><p><b>CONCLUSION</b>Measuring the right ventricular pressure through venous cannula in external jugular vein with homemade PE pipe in mice gets not only a high success rate but also help to save time. Moreover, this method can be popularized easily. It is a good and feasible method for measuring right ventricular pressure in mice.</p>

The essentiality of dynamic contrast enhanced magnetic resonance imaging examination before breast-conserving surgery in breast cancer patients with young women / 中国医师进修杂志

Objective To explore the essentiality of dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) examination before breast-conserving surgery (BCS) in breast cancer patients with young women.Methods The data of DCE-MRI and mammography X-ray in 30 breast cancer patients with young women (＜ 40 years old) who received surgery with pathology-proven were retrospectively analyzed.The detection rate of breast lesion,number,size,extent of invasive and lymph node metastasis of preoperative were observed.The DCE-MRI was assessed accuracy of BCS in accordance with indications for BCS and histopathology results.Results The breast cancer detection rate of DCE-MRI was significantly higher than mammography X-ray [100.00％(30/30) vs.73.33％(22/30),P =0.002].The maximum tumor diameter of DCE-MRI was (3.55 ± 1.50) cm,mammography X-ray was (3.38 ± 1.70) cm,postoperative pathology was (3.51 ± 1.20) cm,and DCE-MRI was closer to postoperative pathology than mammography X-ray.The discovery of additional lesions of DCE-MRI was more than mammography X-ray (P ＜ 0.05).The surrounding tissue invasion of DCE-MRI was detected in 22 cases,mammography X-ray was in 9 cases,postoperative pathology was in 19 cases,there was no significant difference between DCE-MRI and postoperative pathology (P =0.598),there was significant difference between mammography X-ray and postoperative pathology (P =0.010).The number of axillary fossa lymph node metastasis of DCE-MRI was in 127,mammography X-ray was in 48,compared with postoperative pathology (91),there was significant difference (P =0.026,0.034).The detection of microcalcification of mammography X-ray was better than DCE-MRI (7 cases vs.0 case,P =0.001).The number of BCS of postoperative pathology was in 12 cases,DCE-MRI was in 11 cases,mammography X-ray was in 8 cases,DCE-MRI was higher than mammography X-ray,but there was no significant difference (P=0.132).Conclusion DCE-MRI provides comprehensive and accurate information for breast cancer patients with young women for BCS,and it is feasible to assess them using DCE-MRI preoperative for BCS.

Spatio-temporal expression study of phosphorylated 70-kDa ribosomal S6 kinase (p70S6k) in mesial temporal lobe epilepsy / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To determine the spatio-temporal expression of p70S6k activation in hippocampus in mesial temporal lobe epilepsy.</p><p><b>METHODS</b>Temporal lobe epilepsy model was established by stereotaxically unilateral and intrahippocampal injection of kainite acid (KA) in adult male C57BL/6 mice. Latent and chronic epileptogenesis were represented by mice 5 days after KA injection (n = 5) and mice 5 weeks after KA injection (n = 8), respectively. Control mice (n = 5) were injected with saline. Immunohistochemical assays were performed on brain sections of the mice.</p><p><b>RESULTS</b>Hippocampus both ipsilateral and contralateral to the KA injection displayed significantly up-regulated pS6 immunoreactivity in dispersed granule cells in 5-day and 5-week model mice.</p><p><b>CONCLUSION</b>The activation of p70S6k is mainly located in the dentate gyrus in KA-induced mouse model of temporal lobe epilepsy, indicating that the activation may be related with the disperse degree and hypertrophy of granule cells.</p>

Effect of celecoxib on pulmonary hypertension of chronic hypoxia and hypercapnic rats / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To study the effect of celecoxib on chronic hypoxia and hypercapnic pulmonary hypertension.</p><p><b>METHODS</b>SD rats were randomly divided into normal control group (A), hypoxic hypercapnic group (B), hypoxic hypercapnia+ celecoxib group (C). The content of TXB2 and 6-keto-PGF1alpha in plasma and lung were detected by the technique of radioimmunology.</p><p><b>RESULTS</b>(1) Mean pulmonary arteria pressure(mPAP) was significantly higher in rats of B group than those of A group. mPAP was significantly higher in rats of C group than those of B group. Differences of mPAP were not significant in three groups. (2) The content of TXB2 in plasma and lung and the ratio of TXB2/6-keto-PGF1alpha were significantly higher in rats of B group than those of A group. The ratio of TXB2/6-keto-PGF1alpha was significantly higher and the content of 6-keto-PGF1alpha in plasma and lung was significantly lower in rats of C group than those of B group. (3) Light microscopy showed that WA/TA (vessel wall area/total area) and PAMT (the thickness of medial smooth cell layer) were significantly higher in rats of B group than those of A group. WA/TA and PAMT were significantly higher in rats of C group than those of B group. (4) Electron microscopy showed the thickening of vessel wall and the proliferation of collagen fiber in B group and augmentation of smooth muscle cell and abundance of myofilament in pulmonary arterioles in C group.</p><p><b>CONCLUSION</b>Celecoxib can aggravate hypoxic hypercapnia pulmonary hypertension and pulmonary vessel remodeling by increasing the ratio of TXA2/PGI2.</p>

Cyclooxygenase-2 overexpression and vascular endothelial growth factor expression in pharyngeal tissue of patients with OSAHS correlates with angiogenesis / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To reseach the correlations between cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF) expressions and angiogenesis in pharyngeal tissue of patients with obstructive sleep apnea hypopnea syndrome (OSAHS).</p><p><b>METHODS</b>Biopsies were obtained by uvulopalatopharyngoplasty from 40 patients with mild to severe OSAHS. Control specimens of palatopharyngeal and palatoglossal arch mucosa were retreved from 6 patients with chronic tonsillitis and proved have no related disorders. HE was used to observe the changes of pharyngeal tissue, immunohistochemical staining with antibodies against COX-2, VEGF, microvessel density (MVD) (marked with CD34).</p><p><b>RESULTS</b>COX-2 and VEGF mainly expressed at pavement-epithelium and glandular epithelium of pharyngeal tissue, and stronger COX-2 and VEGF expression was found in midrange and severe OSAHS than mild and control group (P < 0.01), so as MVD. COX-2 expression was correlated positively with VEGF expression, and had significant correlation with MVD. VEGF expression had the same correlation with MVD. These three targets had considerable relation with apnea hypopnea index (AHI) and lowest O2 saturation at night.</p><p><b>CONCLUSION</b>There was angiogenesis which had important relationship with hypoxia degree in patients of OSAHS, and COX-2 and VEGF play a crucial role in its development.</p>

Role of beta-catenin in the pathogenesis of mesial temporal lobe epilepsy / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the role of beta-catenin in the pathogenesis of mesial temporal lobe epilepsy.</p><p><b>METHODS</b>Kainic acid-induced rat models of medial temporal lobe epilepsy was established. The expression of beta-catenin in the normal mice and the model mice were detected using Western blot analysis. The expression of beta-catenin at human hippocampus was detected using immunohistochemical analysis and immunofluorescence and compared between patients with non-hippocampal sclerosis temporal lobe epilepsy and those with hippocampal sclerosis epilepsy.</p><p><b>RESULTS</b>The pathologies of model mice were similar with those in mice with hippocampal sclerosis temporal lobe epilepsy, demonstrating that the mice model was successfully established. Western blot analysis showed no significant difference of beta-catenin expression between normal mice and model mice. As shown by immunohistochemical analysis and immunofluorescence, beta-catenin expression in human hippocampus was also not significantly different between patients with temporal lobe epilepsy without hippocampal sclerosis and those with hippocampal sclerosis.</p><p><b>CONCLUSION</b>Beta-catenin may not be involved in the development of hippocampal sclerosis of mesial temporal lobe epilepsy.</p>

Effects of puerarin on proliferation, apoptosis and Kv1.5 gene expression of pulmonary artery smooth muscle cells induced by hypoxia / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the effects of puerarin on proliferation, apoptosis and Kv1.5 gene expression of rat pulmonary artery smooth muscle cells (PASMCs) induced by hypoxia.</p><p><b>METHODS</b>The rat PASMCs were divided into 5 groups: control group, hypoxia group, hypoxia plus puerarin (1 × 10(-5) mol/L) group, hypoxia plus puerarin (1 × 10(-4) mol/L) group and hypoxia plus puerarin (1 × 10(-3) mol/L) group, and cultured at 37°C for 24 h. The proliferation of rat PASMCs was detected by CCK-8 assay and flow cytometry, the activity of caspase-3 was measured with spectrophotometric method, Kv1.5 protein was detected by western blot, Kv1.5 mRNA was detected by real-time PCR.</p><p><b>RESULTS</b>The cell viability and proportion of synthesis phase in control group were 0.940 ± 0.045 and 9.67% ± 1.28%, which were significantly lower than those (1.296 ± 0.034 and 18.19% ± 1.19%) in hypoxia group (P < 0.05). The Caspase-3 activity, Kv 1.5 protein and Kv 1.5 mRNA in control group were 0.1073 ± 0.0113, 0.886 ± 0.038 and 0.0377 ± 0.0031, which were significantly higher than those (0.0664 ± 0.0049, 0.602 ± 0.064 and 0.0108 ± 0.0014) in hypoxia group (P < 0.05). As compared with hypoxia group, the cell viability and proportion of synthesis phase in 3 hypoxia plus puerarin groups significantly decreased, and the Caspase-3 activity, Kv 1.5 protein and Kv 1.5 mRNA in 3 hypoxia plus puerarin groups significantly enhanced (P < 0.05).</p><p><b>CONCLUSION</b>Puerarin could decrease the proliferation and increase the apoptosis induced by hypoxia in rat PASMCs, and the up-regulated expression of Kv1.5 gene may be the mechanism of puerarin effects.</p>

Effects of tetramethylpyrazine on fractalkine and tumor necrosis factor-alpha expression in patients with chronic pulmonary heart disease / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To reveal the relationship of chronic pulmonary heart disease (CPHD) with the chemotactic factor Fractalkine (FKN) and tumor necrosis factor-alpha (TNF-alpha), and to explore the action mechanism of tetramethylpyrazine (TMP) for suppressing pulmonary hypertension.</p><p><b>METHODS</b>Patients with CPHD were randomly assigned to two groups, 19 in Group A and 16 in Group B, and a control group (group C) consisting of 18 healthy adults was setup. Conventional treatment were given to all patients, which consisted of Piperacillin 3. 375 g iv dripping twice a day, Levofloxacin 0.6 g + Ambroxol 60 mg + Doxofylline 0.2 g iv dripping once a day, all for 10-14 days, and acid-base and electrolytes balance in patients were monitored and corrected. At the same time, TMP (trade name: Chuanqing, containing 120 mg of TMP in a 2 mL ampoule) was given additionally to patients in Group B at the dosage of 240 mg/d by adding in 250 mL of normal saline via iv dripping. Serum levels of FKN and TNF-alpha were detected before and after treatment by enzyme-linked immunoassay, and the change of mean pulmonary arterial pressure (mPAP) was measured as well.</p><p><b>RESULTS</b>Before treatment, difference of FKN and TNF-alpha levels between the two patients' groups were insignificant (P > 0.05), but all higher than those in Group C respectively (P < 0.01). While after treatment, the two indices and mPAP levels in Group B were statistically lower than those before treatment, also than those in Group A. Regression analysis showed a positive correlation between TNF-alpha and FKN (r = 0.662, P < 0.001).</p><p><b>CONCLUSIONS</b>A high blood FKN and TNF-alpha expression state exists in CPHD patients, which could be suppressed by TMP, and these suppressive effects may be one of the important mechanisms responsible for the pulmonary arterial pressure lowering action of TMP.</p>

Primary cell culture and identification methods of rat pulmonary arterial smooth muscle cells / 中国应用生理学杂志

<p><b>OBJECTIVE</b>Set up a method to isolate and identify the small pulmonary arterial smooth muscle cells (PASMCs) in vitro.</p><p><b>METHODS</b>In sterile conditions, separated the male SD rat pulmonary artery, digested by collagenase I and cultured primary PASMCs. Measured cell viability; observed by phase contrast microscope; identified by immunocytochemistry and immunofluorescence staining as a label for smooth muscle alpha-actin.</p><p><b>RESULTS</b>PASMCs were identified by morphology and immunocytochemistry, immunofluorescence staining, with the cell viability is over 96.5%. The primary culture could be subcultured after 4-7 days and successfully passaged without change in morphology and growth characteristic.</p><p><b>CONCLUSION</b>This technique has advantage of the method is simple, short cultivate, good reproducibility, the primary cultured PASMCs quantity and the rapid growth.</p>

The effect of chimonin on collagen metabolism of pulmonary arterioles in chronic hypoxic rats / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To study the effect and mechanism of chimonin on pulmonary arterioles I and III type collagen metabolism in pulmonary hypertension rats induced by chronic hypoxic hypercapnia.</p><p><b>METHODS</b>Thirty-six Sprague-Dawley rats were randomly divided into three groups: normal control group(A), hypoxic hypercapnic group(B), hypoxic hypercapnia + chimonin group(C). Collagen I, III and their mRNA, Blood CO concentration (COHb%), activity of HO-1 in blood serum and lung homogenate, content of hydroxyproline in lung homogenate, pulmonary arteriole micromorphometric index were observed.</p><p><b>RESULTS</b>Hypoxic hypercapnic rats's mPAP, Hyr of lung homogenate, content of I type collagen and I type collagen mRNA in pulmonary arterioles, were significantly higher than those in control group, pulmonary vessel remodeling of hypoxic hypercapnic rats was significant, those changes in hypercapnia + chimonin group were significantly lower than those in hypoxic hypercapnic group. Blood CO concentration, activity of HO-1 in blood serum and lung homogenate in rats of hypoxic hypercapnic rats were significantly higher than those of control group, and those of hypercapnia + chimonin group were even higher than hypoxic hypercapnic group (P < 0.01). There was no significant difference in mCAP, content of III type collagen and their mRNA in three groups (P > 0.05).</p><p><b>CONCLUSION</b>Chimonin can reduce pulmonary hypertension and pulmonary vessel remodeling induced by hypoxic hypercapnia through inhibiting proliferation of collagen I, the mechanism maybe is up regulating endogenous carbon monoxide system.</p>

The role of angelica injection on P-selectin and anti-cardiolipin antibodies in acute pulmonary embolism rats / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To study the effect of Chinese medicine, Angelica, injection on the expression of P-, E-selectin and anti-cardiolipin antibody in acute pulmonary embolism rats.</p><p><b>METHODS</b>SD rats were randomly divided into 3 groups: normal control group(Group N), thromboembolism group (Group T), and treatment group of thromboembolism with angelica injection (Group TA). There were three time points in every group: 1 h, 4 h and 8 h. Plasma was detected by P-, with 4% paraformaldehyde, and paraffin embedded sections were detected by immunohistochemistry for P-, E-selectin and anti-cardiolipin antibodies.</p><p><b>RESULTS</b>With HE stain, the inflammatory cells in the lung of rats were relatively rare in every time point in normal control group. In group T and group TA, the inflammatory cells were increasing in every time point in comparison to group N (P < 0.05) and the inflammatory cells were increasing with time in group T. The data revealed that the plasmic level of P-, E-selectin was significantly higher than that in group T1, group T4, group T8 in comparison to the corresponding sub groups of group N (P < 0.05), while it was significantly lower than that in group TA1, group TA4, group TA8 in comparison to the corresponding sub groups of group T (P < 0.05); For the OD value of plasmic anti-cardiolipin antibodies (ACA), no significant difference was observed during was lower expressed by immunohistochemistry.</p><p><b>CONCLUSION</b>Acute pulmonary embolism can lead to infiltration of inflammatory cell in rat lungs. The lung inflammation of acute pulmonary embolism rats can be enhanced probably by the increased release of P-, E-selectin and anti-cardiolipin antibodies, and the enhanced inflammation promotes the release of a series of inflammatory mediators, which exacerbate the injury of lung. Angelica injection relieves the lung inflammation of acute pulmonary embolism rats possibly by inhibiting the expression of P-, E-selectin and anti-cardiolipin antibody, thus playing a role in reducing thrombogenesis.</p>