Objective To investigate calcification of cultured aortic medial cells in vitro and acceleration by 25 hydroxycholesterol or ? glycerophosphate. Methods Aortic medial cells were obtained by explantation, and the calcification was observed by von Kossa staining. Insoluble calcium precipitation in cellular layer was determined by biochemical method , and osteocalcin in the media was analyzed with radioimmunoassay. Results Two different types of primary cells were shown from culture: one with parallel cellular growth and being negative by von Kossa staining, the other cell type formed cellular nodules with positive von Kossa staining. After 28 days of cell passages, cell growth appeared no nodule formation. However, many cellular nodules and positive von Kossa staining were observed in the passaged cells treated with 25 hydroxycholesterol or ? glycerophosphate, and both insoluble calcium 〔(57 80?18 50)?g/pool, (67 50?15 30)?g/pool〕and osteocalcin 〔(0 886?0 063)?g/L, (0 895?0 061)?g/L〕in the medium were significantly increased than that of the untreated cells. Conclusions Cultured aortic medial cells could be divided into two subtypes, one with the characters of smooth muscle cells, the other with the micro vascular pericytes which could calcify the extracellular matrix. 25 hydroxycholesterol and ? glycerophosphate promoted the in vitro calcification, and osteocalcin secretion was increased in the process of calcification of aortic medial, suggesting that osteocalcin might participate in the aortic calcification.

Objective: To determine total phenylethanoid glycoside and acteoside in Plantago Herba to provide reference for evaluating the quality of medicinal materials.Methods: With acteoside as the control sample, a UV visible spectrophotometric method was used to determine total phenylethanoid glycosides in Plantago Herba.An HPLC method was applied to determine acteoside in Plantago Herba , and the conditions were as follows: an ODS2 C 18 (150 mm× 4.6 mm ,5 μm) chromatographic column was used with acetonitrile-0.1% formic acid (13∶87) as the mobile phase at a flow rate of 1.0 ml·min-1 , the detection wavelength was 332nm, the column temperature was 30℃, and the sample volume was 10 μl.Results: The reference solution and the sample solution had the maximum absorption at 332 nm, and the linear relationship was good within the range of 0.003 1-0.155 0 mg·ml-1 (r=0.999 5).The content of total benzene alcohol glycosides in 3 batches of samples was 2.73% , 2.61% and 2.84% , respectively;acteoside over the range of 0.000 6-0.155 0 mg·ml-1 (r=0.999 1) showed a good linear relationship with peak area,the sample recovery was 98.5% and the RSD was 1.6% (n =6), and the acteoside content in 3 batches of samples respectively was 0.54% , 0.51% and 0.56%.Conclusion: The method is simple, accurate and reproducible, and can be used for the determination of total phenylethanoid glycosides and acteoside in Plantago Herba.

Objective To investigate the direct and indirect MRI manifestations of anterior cruciate ligament(ACL)tears and discuss its mechanism and diagnostic value.Methods The study involved 40 patients with ACL tears including 37 males and three females at age range of 16-49 years(mean age of 33 years).Arthroscopy and operation confirmed 28 patients at acute phase of ACL tears,12 at chronic phase of ACL tears,35 with complete ACL tears and five with partial ACL tears.All patients were examined on 1.5T Siemens MRI scanner,with SE,TSE or Medic sequence in multiple directions.The direct and indirect MRI manifestations of ACL tears were retrospectively and statistically analyzed by employing three experienced doctors.Results Among 28 patients with acute tears,the direct MRI signs included signal interruption or non-consecutive(86%),uneven signal(64%)and swelling and thickening ligament(36%).While in 12 patients with chronic teats,the MRI signs manifested swelling ligament(92%),signal interruption or non-consecutive(75%)and uneven signal(58%).The sign of thickening ligament was mainly seen at chronic phase of ACL tears(P ＜0.01).Of 35 patients with complete ACL tears,80% occurred in the middle part of ACL,with low incidence of upper and lower parts.All five patients with partial ACL tears occurred in the anteriomedialis bundle.The indirect MRI signs of ACL tears included 7-shaped deformity of posterior cruciate ligament(PCL)(34 patients,85%),meniscus exposure(26 patients)including 16 with lateral meniscus exposure(62%),bone injury(15 patients)including segond fracture(8 patients,53%),widened joint space(9 patients)including 78% at chronic phase,and tibia antelocation(23 patients,57%).Among 40 patients,37 patients were diagnosed correctly preoperatively,with accuracy rate of 92%.While three patients with partial ACL tears were missed diagnosis preoperatively,with inaccuracy rate of 8%.Conclusion ACL tear is relatively easy to identify preoperatively according to direct and indirect MRI signs in combination with trauma history.

Objective To observe the surface area temperature of dysmenorrhea rats with cold stagnation syndrome; To compare the different effects of Sanyinjiao (SP6) and Guanyuan (RN4).Methods Forty female SD rats were randomized into control group, model group, Sanyinjiao (SP6) group, and Guanyuan (RN4) group, 12 rats in each group. Whole body freezing method combined with estradiol benzoate injection was used to establish models. Sanyinjiao (SP6) group and Guanyuan (RN4) group received moxibustion at corresponding points for 3 times after modeling. Infrared thermal imaging was used to measure the skin temperature at the surface projection area of Sanyinjiao (SP6) and Xuehai (SP10) and Sanyinjiao-Xuehai lines before and after moxibustion.Results Compared with control group, the temperature gap between double sides of Sanyinjiao-Xuehai lines significantly increased in model group 5-30 min after moxibustion (P<0.05). 30 min after moxibustion, the temperature of right Sanyinjiao significantly decreased in other three groups (P<0.01). Compared with model group, the temperature gap between double sides of Sanyinjiao-Xuehai lines significantly decreased after 5-30 min in Sanyinjiao group (P<0.05), while Sanyinjiao-Xuehai lines significantly decreased after 10-30 min in Guanyuan group (P<0.05). Comparison between two moxibustion groups, the influence of Sanyinjiao group to temperature gap between double sides of Sanyinjiao-Xuehai lines was earlier than Guanyuan group (P<0.05).Conclusion Moxbustion can decrease the temperature gap between double sides of Sanyinjiao-Xuehai lines and ease the imbalance. And the influence of moxbustion SP6 to temperature gap between double sides of Sanyinjiao-Xuehai lines is earlier than moxibustion RN4.

Sepsis-associated encephalopathy(SAE) is a diffuse brain dysfunction caused by sepsis.The main clinical manifestations are abnormal mental state, high mortality and poor prognosis.At present, there is no unified diagnostic standard for SAE.The exclusion diagnosis is mainly based on clinical symptoms and signs, combined with laboratory examination and imaging auxiliary examination.Among them, brain magnetic resonance imaging and quantitative electroencephalography can early detect brain dysfunction and predict the prognosis of children, which play an important role in the early diagnosis and prognosis assessment of SAE.Cerebral oxygen monitoring can dynamically reflect the changes of brain function and can be used for long-term monitoring of children with severe brain function injury.SAE is closely associated with poor prognosis, and mortality will increase with the extention of hospitalization.Therefore, early identification of SAE is of great significance to reduce mortality.

Objective Infrared thermal imaging can be applied to the diagnosis and auxiliary diagnosis of some diseases . The aim of this study is to explore acupuncture-induced changes in skin temperature in acupoint areas and whether skin temperature in -creases or decreases in the acupoint areas along meridians . Methods Thirty two female SD rats were randomly and equally divided into four groups:saline control,cold congealing and dysmenorrhea model , Sanyinjiao (SP6), and Guanyuan( CV4).Models were es-tablished in the latter three groups by subcutaneous injection of estradiol benzoate at 0.5 mg for 10 successive days and , 1hour after the last administration , intraperitoneal injection of oxytocin at 2 U, followed by exposure of the rats to-25℃in a freezer 4 hours a day for 5 days.Meanwhile , the control rats received normal saline only and were not exposed to low temperature .Infrared thermal imaging was used to measure the skin temperature at the acupoint areas of SP6, Xuehai (SP10), and CV4 before and at 0, 5, 10, 20, 30, 40, 50 and 60 min after needling . R esults At 0 to 5 min after nee-dling, the skin temperature of the left SP6 and right SP10 was signifi-cantly decreased in both the SP6 and CV4 groups ( [ -0. 56 ± 0.22]℃and [-0.48 ±0.11]℃, P<0.01), and so was that of the right SP10 ([ -0.64 ±0.21]℃ and [ -0.45 ±0.13]℃, P<0.05).At 5 to 10min, the skin temperature of the right SP6 and SP10 was markedly increased in the SP6 group ([-0.49 ±0.35]℃and [-0 .18 ±0.20]℃, P<0.01), and so was that of the right SP6 in the SP6 group at 20 to 30 min ([ -0.14 ±0.25]℃) as compared with the model and CV4 groups (P<0.01).At 30 to 40 min, the skin temperature of the right SP10 was remarkably elevat-ed in the SP6 group ([ -0.03 ±0.11]℃) in comparison with the model group (P<0.01).No significant differences were observed in the skin temperature of the left SP10 and CV4 at different time points among the four groups (P>0.05). Conclusion The skin temperature of SP6 and SP10 can be regulated by needling both the acupoints of SP 6 and CV4.The increase in the skin temperature of the right SP6 and SP10 in the SP6 group and no change in the CV 4 group indicated dynamic temperature changes in the acupoint area along the meridian after needling.

Objective To observe the analgesic effect of acupuncture at Guanyuan (CV4) and its effect on vasomotor substances in rats with dysmenorrhea due to coagulated cold syndrome. Method The coagulated-cold dysmenorrhea rat model was developed by Estrodiol benzoate and Oxytocin injectin plus physical freezing. The writhing response (writhing latency, writhing frequency, and writhing score) was observed, and the contents of TXB2 and 6-keto-PGF1a were detected by using enzyme-linked immunosorbent assay (ELISA). Result Compared with the saline water group, the writhing latency was significantly shortened, the writhing frequency was significantly increased, and the writhing score was more significantly increased in the model group (P<0.01);compared with the model group, the writhing latency was significantly prolonged, the writhing frequency was decreased, and the writhing score was significantly lower in the acupuncture group (P<0.05, P<0.01). Compared with saline water group, the content of plasma 6-keto-PGF1a was significantly lower (P<0.05) and the content of plasma TXB2 showed an increasing tendency (P>0.05) in the model group. Compared with the model group, the content of plasma 6-keto-PGF1a showed an increasing tendency (P>0.05) and the content of plasma TXB2 showed a decreasing tendency (P>0.05) in the acupuncture group. Conclusion The vasomotor substances are obviously disordered in the blood of cold-coagulated dysmenorrhea rat models. Acupuncture at Guanyuan can improve the writhing response and release pain, and meanwhile positively regulate the vasomotor substances such as TXB2 and 6-keto-PGF1a. The vasomotor substances are plausibly one of the major substances in the action of acupuncture in preventing and treating dysmenorrhea.

Objective To explore the effect of different acupuncture stimuli on uterine micro-circulation of dysmenorrheal rats with cold stagnation syndrome. Methods Totally 32 three-month old female SD rats in diestrus were randomly divided into saline control group, model group, A stimuli group, and B stimuli group, 8 rats in each group. Model group and treatment groups were given whole body freezing combined with estradiol benzoate injection method to establish models. A stimuli group was given deep puncture with manipulation, while B stimuli group was treated by shallow puncture without manipulation. Diameter of uterine capillary,micro-vessel, TXB2, and 6-keto-PGF1αlevels were observed in each group. Results Compared with the saline group, capillary diameter in model group was significantly reduced at 5, 10, 20, 30 min time point (P<0.01);micro-vascular diameter was significantly reduced at 5, 10, 20, 30, 40 min time point (P<0.01);plasma 6-keto-PGF1α levels decreased (P<0.01);TXB2/6-keto-PGF1αincreased significantly (P<0.01). Compared with model group, capillary diameter in A stimuli group enlarged at 5, 10, 20, 30 min time point (P<0.05), micro-vascular diameter dilated at 5, 10, 20, 30, 40 min time point (P <0.01), plasma 6-keto-PGF1α level increased (P <0.05), TXB2/6-keto-PGF1αdecreased significantly (P<0.05);micro-vascular diameter in B stimuli group dilated at 20, 30 min time point (P<0.05), plasma TXB2/6-keto-PGF1α decreased significantly (P<0.05). Compared with B stimuli group, capillary diameter in A stimuli group dilated at 5, 10, 20, 30 min time point (P<0.05) and micro-vascular diameter dilated at 5, 10, 20, 30, 40 min time point significantly (P<0.01). Conclusion Dysmenorrheal rats with cold stagnation syndrome show obvious disorder of the uterus micro-circulation and circulation related substances. Both A and B acupuncture stimuli improved uterus micro-circulation of dysmenorrheal rats with cold stagnation syndrome, and its mechanism may be related to the recovery the balance between TXB2 and 6-keto-PGF1α.

Objective:To investigate the correlation between serum uric acid level and heart rate variability (HRV) in patients with acute ischemic stroke.Methods:Patients with acute ischemic stroke admitted to the Department of Neurology, the First Affiliated Hospital of Nanjing Medical University from July 2017 to July 2019 were enrolled retrospectively. They were divided into 4 groups (Q1, Q2, Q3, and Q4) according to the sex-specific serum uric acid quartiles. The demographic data, baseline clinical data, laboratory indicators and HRV recorded by 24-h dynamic electrocardiogram in each group were compared. The independent correlation between HRV time domain parameters RR (normal-to-normal, NN) interval standard deviation (standard deviation of the NN intervals, SDNN), RR interval mean standard deviation (standard deviation of the average NN intervals, SDANN) and serum uric acid level was analyzed through multiple linear regression model.Results:A total of 141 patients were enrolled in the study, including 100 males (70.92%), aged 61.55±12.92 years. The median baseline National Institutes of Health Stroke Scale (NIHSS) score was 2 (interquartile range: 1-4), and the serum uric acid level was 302.87±73.48 μmol/L. There were significant differences in the NIHSS score ( P<0.034), the proportion of patients with hypertension ( P=0.015), the estimated glomerular filtration rate ( P<0.001), and the HRV time domain parameters (all P<0.001) between each quartile group. The proportion of patients with low HRV (SDNN <100 ms) in the lowest serum uric acid level quartile group was significantly higher than that in other quartile groups ( P<0.05). Multiple linear regression analysis showed that serum uric acid levels were significantly and positively correlated with SDNN (standardized β=0.321, P<0.001) and SDANN (standardized β=0.266, P=0.001). Conclusions:The low level of serum uric acid in patients with acute ischemic stroke indicates that HRV is low, which may suggest that the mechanism of oxidative stress is involved in the decrease of HRV in patients with acute ischemic stroke.

Objective:To investigate the neutrophil to lymphocyte ratio (NLR) and clinical manifestations, imaging manifestations and plasma D-dimer in cerebral venous sinus thrombosis (CVST) patients with headache as the first symptom.Methods:CVST inpatients with headache as the first symptom admitted to the First Affiliated Hospital of Nanjing Medical University from January 2012 to December 2019 were enrolled retrospectively as case group. In the same period, patients with similar headache symptoms and healthy subjects were included consecutively as control group. The differences of demographic data, biochemical indexes, blood routine indexes and plasma D-dimer were compared among the three groups. Multivariate logistic regression analysis was used to identify the independent correlation between NLR and plasma D-dimer and CVST with headache as the first symptom. According to the median NLR, patients with CVST were divided into low NLR group and high NLR group, and the differences in clinical manifestations, imaging manifestations and plasma D-dimer were compared between the two groups. Multiple linear regression analysis was used to determine the independent correlation between the reciprocal of NLR and plasma D-dimer and clinical features. Results:A total of 51 CVST inpatients with headache as the first symptom, 54 headache controls and 53 healthy controls were included in the study. There were significant differences in triglyceride ( P=0.002), fasting blood glucose ( P=0.044), serum albumin ( P=0.013), uric acid ( P=0.010), plasma D-dimer ( P<0.001), proportion of D-dimer negative patients ( P<0.001), mean corpuscular volume ( P=0.015), white blood cell count ( P<0.001), neutrophil count ( P<0.001), lymphocyte count ( P=0.005), monocyte count ( P=0.004), mean platelet volume ( P=0.010), platelet distribution width ( P=0.029), NLR ( P<0.001) and lymphocyte to monocyte ratio ( P<0.001) among the three groups. Post-hoc analysis showed that plasma D-dimer and NLR in the case group were significantly higher than those in the two control groups (all P<0.001). Multivariate logistic regression analysis with the headache control group and the healthy control group as references showed that NLR and plasma D-dimer were significantly and independently correlated with CVST with headache as the first symptom (all P<0.05). According to the median NLR (4.817 6), the case group was divided into low NLR group and high NLR group. The comparison showed that the onset time ( P=0.041), the proportion of patients with headache as the only symptom ( P=0.016) and the proportion of plasma D-dimer negative patients ( P=0.002) in the low NLR group were significantly higher than those of the high NLR group, and the proportion of patients with headache complicated with disturbance of consciousness ( P=0.032), plasma D-dimer ( P<0.001) and fasting blood glucose ( P<0.001) were significantly lower than those in the high NLR group. With the reciprocal of NLR as the dependent variable, multiple linear regression analysis showed that the reciprocal of NLR was independently negatively correlated with plasma D-dimer ( R=-0.521, P<0.001), and independently positively correlated with headache as the only symptom ( R=0.280, P=0.049). Conclusions:NLR was significantly increased in CVST patients with headache as the first symptom. It might help early diagnosis and clinical feature prediction of CVST patients.