OBJECTIVEThe content of vindoline, catharanthine and vinblastine in the root, stem, leaf, flower and fruit of Catharanthus roseus at various developmental stages were determined, and the biomass allocation was also determined to find the best harvest time.

METHODThe content of vindoline, catharanthine and vinblastine in the root, stem, leaf, flower and fruit of C. roseus were determined by HPLC.

RESULTThe content of these alkaloids were influenced by season and it varied in the different tissues of the plant. The content of vindoline and catharanthine in the leaves were the highest, and there was no vindoline detected in the root, but the content of vinblastine in the flower was the highest; the content of vindoline and catharanthine reached the maximum between the August and September, and the content of vinblastine reached the highest after the September. The biomass was the highest in the initial stage of September.

CONCLUSIONThe best harvest time was in the initial stage of September.

Catharanthus ; chemistry ; growth & development ; metabolism ; China ; Chromatography, High Pressure Liquid ; Flowers ; chemistry ; growth & development ; metabolism ; Plant Extracts ; analysis ; metabolism ; Plant Leaves ; chemistry ; growth & development ; metabolism ; Plant Roots ; chemistry ; growth & development ; metabolism ; Seasons ; Vinblastine ; analogs & derivatives ; analysis ; metabolism ; Vinca Alkaloids ; analysis ; metabolism

Objective: To observe the effects of blueberry and nuclear expression of transcription factor-кb (NF-кb) p65 in an experimental rat model of liver fibrosis. Methods: Forty-five Sprague-Dawley rats were randomly divided into isotonic saline control group (A); model group (B); blueberry juice prevention group (C, 15 g/kg); dan-shao-hua-xian capsule prevention group (D, 1 g/kg); and blueberry juice + dan-shao-hua-xian capsule prevention group (E). Rat liver fibrosis model was established by covalent compound carbon tetrachloride (CCl₄). Each prevention group was given the corresponding dose of blueberry juice or (and) dan-shao-hua-xian capsule, and the rats were sacrificed after 8 weeks. The degree of liver fibrosis was evaluated by hematoxylin and eosin stain. A liver tissue of NF-κBp65 was detected by immunohistochemical method. The NF-κBp65 protein expression of liver tissue and transforming growth factor (TGF) β1 was detected by Western blot. Data of multiple groups were compared by one-way analysis of variance, and rank sum test. Results: Immunohistochemistry detected that TGFβ1 protein was mainly expressed in mesenchymal origin of hepatic stellate cells. The expression level of group A (3.75 ± 1.67) was low, while those of group B (9.00 ± 2.07), C (7.33 ± 1.00), D (6.00 ± 1.51), and E (3.5 ± 1.41) were high. However, the expression level of TGF-β1 protein in hepatic tissues of group B was significantly higher than that of group C, D and E [group E: 3.5 ± 1.41, F = 18.350, P < 0.05]. In addition, group D was higher than group E (P < 0.05). The expression of NF- kappa Bp65 protein was very complex, and the expression patterns in different groups were different (Statistical calculation of experimental data were based on expression of liver cells). Compared with group B (4.37 ± 2.13), the relative expression levels of NF-κBp65 protein in-group A (0.46 ± 0.25), group C (2.76 ± 1.01), group D (2.13 ± 1.51), group E (1.88 ± 0.99) were significantly decreased (F = 27.490, P < 0.05), and the expression trend was consistent with TGFβ1 protein. Western blot detected NF-κBp65 protein in liver tissues of rats. Compared with group A, levels in groups B, C, D and E were significantly increased (F = 96.983, P < 0.05), and groups C, D and E were significantly lower. The E group was significantly lower than the C group (F = 96.983, P < 0.05), and the degree of hepatic fibrosis was lower in each prevention group than in the B group (T = 24.1, P < 0.05). Conclusion Blueberries have preventive effect on CCl4-induced hepatic fibrosis in rats, and its preventive mechanism may inhibit the expression and activation of NF-κBp65 in hepatocytes, thereby reducing TGFβ1- mediated production or activation.

At present, there are a series of problems in the clinical teaching of postgraduates, such as insufficient integration of professional teaching and ideological and political education, insufficient understanding of teachers and students, insufficient exploration of ideological and political elements, and immature construction of teaching system. This paper mainly starts from three aspects: improving the teaching system and mechanism, building a disease-based curriculum ideological and political chain and teaching implementation strategies, organically combines different professional curriculum contents with ideological and political education, and carries out preliminary exploration and practice in the construction of ideological and political teaching system of orthopedic clinical curriculum, so as to guide students to establish correct values, outlook on life and world outlook. It provides useful practical experience for improving the quality of clinical teaching and promoting the reform and innovation of graduate teaching.

Objective To investigate the promotion effect of Danhong injection (DH) on brain-derived neurotrophic factor (BDNF) expression in Schwann cells (SCs) of SD rats.Methods In experiment of SCs apoptosis induced by advanced glycation end products (AGEs),SCs were divided into control group,AGEs treatment group and DH+AGEs treatment group; 48 h after each treatment,the SCs count was compared.In experiment of DH affecting mRNA and protein BDNF expressions in SCs,real time-PCR and Western blotting were used.In the experiment of DH combined with different inhibitors (Calphostin C,LY294002,H89,U0126,FR180204 and SB203580) affecting mRNA BDNF expression in SCs,real time-PCR was used.Results The number of SCs in AGEs treatment group was significantly decreased than that in the control group,but that in DH+AGEs treatment group was statistically increased than that in AGEs group (P＜0.05).The mRNA and protein expressions of BDNF in the DH treatment group were significantly increased than those in the control group (P＜0.05).As compared with DH group,DH+Calphostin C treatment group had significantly decreased BDNF mRNA expression (P＜0.05); BDNF mRNA expression in the DH+U0126,DH+FR180204 and DH+SB203580 treatment groups was significantly decreased as compared with that in the DH treatment group (P＜0.05).Conclusions DH could effectively inhibit SCs apoptosis induced by AGEs and significantly promote BDNF expression;protein kinase C (Calphostin C) and mehtyl ethyl ketone (U0126)/extracellular regulated protein kinases (FR180204EK)/p38 (SB203580) may be the important signaling transconduction pathways for BDNF expression.

Objective:To investigate the clinical characteristics of cerebellopontine angle (CPA) tumors with hemifacial spasm (HFS) or trigeminal neuralgia (TN) as first symptoms.Methods:From May 2015 to November 2018, 70 patients with CPA tumors found in preoperative imaging examination or during operation and with TN or HFS as first symptoms, were enrolled into experimental group; 66 patients with CPA tumors without TN or HFS were chosen as control group. Gender, age, course of the disease, pathological types of the tumors, and tumor sizes were retrospectively analyzed and compared between the two groups.Results:In the experimental group, 54 patients were with TN as first symptom and 16 were with HFS as first symptom. The ratio of male to female in the experimental group was about 1∶2.18, and the difference was not statistically significant as compared with that in the control group (1∶3.00, P>0.05). The median course of disease in the experimental group was 36 (24, 84) months, which was significantly longer than that in the control group (9 [2, 24] months, P<0.05). In the experimental group, 34 patients (48.6%) had cholesteatoma, 24(34.3%) had meningioma, 8 (11.4%) had acoustic neuroma, 2 (2.9%) had lipoma, and 2 (2.9%) had jugular bulb tumor. In the control group, 39 patients (59.1%) had acoustic neuroma, 18 (27.3%) had meningioma, 5 (7.6%) had cholesteatoma, 3 (4.5%) had jugular bulb tumor, and one (1.5%) had lipoma. The tumor size in the experimental group (2.00 [1.60, 4.25] cm 2) was significantly smaller than that in the control group (5.60 [4.00, 8.50] cm 2, P<0.05). In the experimental group, 53 patients (75.7%) were found to have offending arteries compressing on corresponding cranial nerves during surgery; the offending artery of patients with TN was superior cerebellar artery; that of 14 patients with HFS was anterior inferior cerebellar artery and that of 2 patients with HFS was posterior inferior cerebellar artery. All patients with confirmed offending arteries underwent microvacular decompression (MVD) after tumor resection. The patients in the experimental group were followed up for 2-48 months after surgery, and the TN or HFS symptoms disappeared. Conclusions:The female patients with CPA tumors with TN or HFS as first symptoms are more common, and TN is far more common than HFS. Cholesteatoma is the most common tumor. As compared with patients with CPA tumors without TN or HFS, CPA tumors patients with TN or HFS have longer course of disease and smaller tumor volume. Most patients have offending vascular compression on the corresponding cranial nerves, tumor resection+MVD is safe and effective treatment for these patients.

Objective:To investigate the clinical application value of frameless stereotactic electroencephalography (SEEG) electrode implantation assisted by domestic Remebot robot in patients with drug-refractory epilepsy.Methods:Ten patients with medically-refractory epilepsy, admitted to and accepted SEEG electrode implantation assisted by domestic Remebot robot in our hospital from October 2021 to March 2022, were chosen in our study. Epileptogenic focuses of these patients were determined. Bone Fiducial Markers were used for registration during surgery, and the registration error and implantation time of each electrode were recorded in detail. Thin-layer CT scan was performed immediately after surgery to observe the occurrence of complications. Three-dimensional reconstruction of all implanted electrodes was performed postoperatively, which was fused with the preoperative surgical planning paths and targets, and the electrode entry point error and target error were calculated.Results:SEEG electrodes were implanted in all 10 patients with the assistance of frameless surgical robot, and the resection areas were accurately determined according to the results of SEEG records. The registered error of these 10 patients was (0.24±0.03) mm. A total of 97 SEEG electrodes were implanted, and the average effective implantation time of each electrode was (6.3±1.2) min. After the fusion of three-dimensional electrode reconstruction with preoperative surgical plan, the mean entry point error of 97 electrodes was (1.9±1.2) mm, and the mean target error was (2.1±1.2) mm. Immediate postoperative head CT showed that none of the 10 patients had surgical complications such as intracranial hemorrhage or severe pneumoencephalos.Conclusion:The SEEG electrode implantation assisted by Remebot frameless robot is safe, accurate, and effective, which can meet the clinical needs.

Objective:To study the efficiency and safety of domestic Remebot robot (neurosurgical positioning and navigation system) in the resection of craniocerebral microlesions and functional area lesions.Methods:Twelve patients accepted resection of craniocerebral microlesions and functional area lesions in our hospital from October 2021 to March 2022 were chosen in our study; navigation, localization and resection of craniocerebral microlesions and functional area lesions were performed with assistance of Remebot robot. Surface Markers were used for optical registration during the surgery, and the registration error was recorded in detail. Scalp incision and bone flap boundary were designed by laser navigation. Postoperative head CT and MRI scan were performed to observe intracranial hemorrhage and lesion resection. The postoperative image was fused with the preoperative surgical plan to calculate the deviation between the planned and actual flap centers.Results:The patient registration error was 0.6-1.5 mm, with an average of (0.9±0.12) mm. The registration error of the manipulator was 0.09-0.12 mm. Preoperative and postoperative image fusion showed that the deviation between the planned and actual flap centers was 1.4-4.5 mm, with an average of (1.9±1.1) mm. All surgical procedures were performed with robot laser navigation to find the lesion boundary along the planned path at one time, which was less destructive to the cortex and surrounding tissues. Postoperative head CT or MRI examination showed that all lesions were completely resected. No serious complications occurred in these 12 patients.Conclusion:Domestic robot can provide precise localization and navigation function for the resection of craniocerebral microlesions and functional area lesions, improving the surgical efficiency and ensuring the safety of surgery.

Objective To investigate the effects of Rehmanniae Radix before and after processing on the intestinal flora of rats with kidney yin deficiency syndrome.Methods SD rats were randomly divided into blank group,model group,probiotic group(0.35 g·kg-1),high-/medium-/low-dose groups of Rehmanniae Radix Praeparata(3.5,1.75,0.875 g·kg-1),and high-/medium-/low-dose groups of Rehmanniae Radix(3.5,1.75,0.875 g·kg-1),with 9 rats in each group.Except for the blank group,rats in each group were injected intramuscularly with Dexamethasone Sodium Phosphate Injection(0.35 mg·kg-1)once a day for 21 days.The drug was administered by gavage once a day on the seventh day of modelling for 14 days.The adrenal histopathological changes were observed by using HE staining;the levels of serum cyclic adenosine monophosphate(cAMP),cyclic guanosine monophosphate(cGMP),corticotropin-releasing hormone(CRH),adrenocorticotropic hormone(ACTH),and corticosterone(CORT)were detected by ELISA;and the levels of short-chain fatty acids in the feces and changes in the diversity of intestinal flora were detected by a targeted metabolomic approach in conjunction with 16SrRNA sequencing in the rats in each group.Results(1)Compared with the blank group,the body mass of rats in the model group was significantly decreased on days 7,14 and 21(P＜0.05,P＜0.01);serum levels of cAMP,CRH,ACTH,CORT and the cAMP/cGMP ratio were all significantly increased(P＜0.01),and the cGMP content was significantly decreased(P＜0.01);and the adrenal cortex was thinned,with the boundaries of various layers of the cortex unclear.Compared with the model group,the body mass of rats in the Rehmanniae Radix Praeparata administration group on day 21 were all significantly increased(P＜0.05,P＜0.01),and the serum content of cAMP,CRH,ACTH,CORT and the cAMP/cGMP ratio were all significantly decreased(P＜0.05,P＜0.01),and the content of cGMP was significantly increased(P＜0.05,P＜0.01)in the rats;body mass of rats in the Rehmanniae Radix administration group did not change significantly(P＞0.05),CRH and CORT contents in serum of rats in the high-dose group of Rehmanniae Radix were significantly reduced(P＜0.01),and ACTH contents in serum of rats in the medium-dose group of Rehmanniae Radix were significantly reduced(P＜0.05);the adrenal cortex of rats in all the administration groups were improved,in particular,the thickening of the adrenal cortex layers was obvious in the Rehmanniae Radix Praeparata group,and the improvement effect was superior to that in the Rehmanniae Radix group.(2)Compared with the blank group,the difference of Coverage index was not statistically significant(P＞0.05),and the coverage of each group was good;the abundance index(Sobs,Ace,Chao)and diversity index(Shannon)of the model group were significantly increased(P＜0.01),and the Simpson index was significantly decreased(P＜0.01).Compared with the model group,Sobs index was significantly decreased in the medium-and high-dose groups of Rehmanniae Radix Praeparata(P＜0.05),Chao index was significantly decreased in the administered groups of Rehmanniae Radix Praeparata and high-dose group of Rehmanniae Radix(P＜0.05,P＜0.01),and Simpson index was significantly increased in the high-dose group of Rehmanniae Radix Praeparata(P＜0.05).The changes of Rehmanniae Radix on the richness and diversity of intestinal microbial community in kidney yin-deficient rats were small,while Rehmanniae Radix Praeparata could better maintain the stability of the richness and diversity of intestinal microbial community in kidney yin-deficient rats.(3)Compared with the blank group,the abundance of phylum firmicutes in the feces of the model group was significantly decreased,while the abundance of bacteroides and actinomycetes was significantly increased.The abundance of Lactobacillus was significantly decreased(P＜0.01),while the abundance of norank_f__Muribaculaceae and Bifidobacterium was significantly increased(P＜0.01).Compared with the model group,the trend of recovery of bacterial abundance in the probiotic group and the high-dose group of Rehmanniae Radix Praeparata was more similar to that of the blank group,which showed that it had the best regulating effect on the ratio of bacterial flora;the abundance of lactobacillus in all administered groups was increased,with that of the probiotic group was significantly increased(P＜0.01);the abundance of norank__f__Muribaculaceae and Bifidobacterium were all decreased,among which the probiotic group and the medium-and high-dose groups of Rehmanniae Radix Praeparata were significantly decreased(P＜0.01),and the effect was significantly superior to that of Rehmanniae Radix.The COG functions of the samples in each group were mainly focused on amino acid transport and metabolism,carbohydrate transport and metabolism,translation,ribosomal structure and biogenesis,replication,recombination and repair,but the abundance information of each function was different between groups,which may be due to the differences caused by dysbiosis of intestinal flora.(4)Compared with the blank group,the levels of acetic acid,butyric acid and propionic acid in the faeces of rats in the model group were significantly decreased(P＜0.05,P＜0.01),and the level of isobutyric acid was significantly increased(P＜0.01).Compared with the model group,the levels of acetic acid,butyric acid and propionic acid in the faeces of rats in the probiotic group and the low-,medium-and high-dose groups of Rehmanniae Radix Praeparata were significantly increased(P＜0.05,P＜0.01)and the levels of isobutyric acid were significantly decreased(P＜0.05,P＜0.01);although the above indexes in the Rehmanniae Radix group were improved,the difference was not statistically significant(P＞0.05).Conclusion The enhanced therapeutic effect of Rehmanniae Radix after processing on rats with kidney yin deficiency syndrome may be related to its adjusting effect on intestinal flora.

Objective: To observe the efficacy and safety related measures by blocking mother-to-child transmission of hepatitis B virus with high viral load and HBeAg positivity during pregnancy in Guizhou province. Methods: Outpatient and inpatient cases of the Department of Infectious Diseases and Obstetrics of Guizhou Medical University Affiliated Hospitals from May 2016 to July 2017 were retrospectively divided into intervention group, non-intervention group and non- hepatitis B pregnant women group; with 75 cases in each group. HBsAg and HBeAg were positive in the intervention group. Pregnant women with HBV DNA ≥10⁶ IU/ml were treated with anti-HBV therapy for 24 to 28 weeks of gestation until delivery. According to oral drugs, they were divided into tenofovir (TDF) group or telbivudine (LDT) group, non-intervention group (HBsAg and HBeAg positive), HBV DNA positive pregnant women, pregnant women with no anti-HBV drugs, non-hepatitis B pregnant women (normal pregnant women without HBV infection). Infants and young children born to the three groups of women were immunized with the national viral hepatitis B action plan. The gestational weeks and Apgar scores at birth, delivery mode, feeding mode, sex and 7-months-old age were observed and counted. Serum hepatitis B markers (HBVM) and HBV DNA were quantitatively detected. HBVM was detected by time-resolved fluorescence immunoassay (TRFIA), and HBV DNA was detected by real-time PCR (FQ-PCR). The changes of liver parameters, HBsAg, HBeAg, HBV DNA, adverse drug reactions and treatment response of pregnant intervention group before medication (12-24 weeks of gestation), 4 weeks of medication (28-32 weeks of gestation), 36-40 weeks of gestation (36-40 weeks of gestation) were statistically calculated. A t-test was used to compare the data between the measurements. Data measurements within the groups were analyzed using rank -sum test. Results: In the intervention group, therapeutic medications showed no differences in demographic and clinical characteristics between TDF group and LDT group, including liver parameters, HBsAg, HBeAg and log10HBV DNA level. Compared with pre-treatment (TDF group: 4.84 ± 2.01; LDT group: 5.08 ± 1.99), TDF and LDT were significantly lower at the end of pregnancy (TDF group: 3.06 ± 0.66; LDT group: 3.51 ± 1.20). P < 0.05); and the treatment response rate was 100%. There were no serious adverse events in the intervention group. Infants and young children (7-months-old) in the intervention group had negative HBsAg, HBeAg and HBV DNA. The mother-to-child transmission rate of HBV was zero, with blocking rate of 100%. In addition, both infants and young children had different degrees of hepatitis B protective antibodies (anti-HBs, M: 144.33), and their antibody titers were higher than that of non-intervention group (anti-HBs, M: 65.91) and non-hepatitis B pregnant women (anti-HBs, M: 58.43). The difference was statistically significant (P < 0.05), and there was no significant correlation between the use of antiviral and the way of delivery and feeding. Outcomes of mother-to-child transmission of HBV infection in infants and young children (7-months-old) delivered by three groups of pregnant women in the non-intervention groups had 20.0% (15/75)/ 17.3% (13/75) HBsAg/HBeAg positivity rate, and 17.3% (13/75) HBV DNA positivity rate. Overall, mother-to-child transmission rate of HBV infection was 20% (15/75). Furthermore, the relationship between mother's HBV DNA load and infant HBV infection in the non-intervention group showed mother's HBV DNA ≥10⁶ IU/ml. Conclusion In the non-intervention group, mother-to-child transmission of HBV occurred, and infected mothers HBV DNA was ≥106 IU/ml before delivery. This suggests that HBeAg positive and high load HBV DNA replication were independent risk factors for mother-to-child transmission of hepatitis B. Therefore, prenatal drug intervention and postpartum standard immune blockade are necessary for high-risk pregnant women with hepatitis B to achieve zero mother-to-child transmission of hepatitis B in real- clinical practice.