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Objective:To investigate the rationality of nerve-plane sparing radical hysterectomy (NPSRH) for cervical cancer by observing the anatomical and histological characteristics of pelvic autonomic plane based on fresh cadaver.Methods:From October 2015 to September 2020, 14 fresh female cadavers were anatomically and histologically studied in the Laboratory of Anatomy and Embryology Department, Peking Union Medical College, Chinese Academy of Medical Sciences. The median age of the specimens was 79 years (range: 67 to 92 years). Twenty-eight hemi-pelvic specimens were obtained from 14 fresh female cadavers. NPSRH procedures were simulated in 8 hemi-pelvic cavities to prove its feasibility. Detailed dissection was conducted to recognize nerve plane and to observe the distribution of pelvic nerves in 10 hemipelvis. In the other 10 hemipelvis, whole parametrium tissue was taken from the crossing of ureter and the uterine artery to the ureterovesical entrance and be embedded, then continuous section was performed, and was stained by hematoxylin-eosin staining (HE) to observe the relationship of nerves and vessels. Immunohistochemical staining of S100, tyrosine dehydrogenase (TH), and vasoactive intestinal peptide (VIP) were performed to count and distinguish sympathetic and parasympathetic nerves, respectively.Results:(1) The pelvic autonomic nerve-plane was completely preserved in 7 of 8 hemipelvis by simulating NPSRH. (2) After detailed dissection in 10 hemipelvis, it was found that hypogastric nerve, pelvic splanchnic nerve, and their confluence of inferior hypogastric plexus were distributed in a planar statelocating in the ureteral mesentery and its caudal extension. This nerve plane showed a cross relationship with deep uterine vein and its branches. The bladder branches and vesical venous plexus were closely related to the inferior hypogastric plexus. The middle vesical vein and inferior vesical vein were intact in 7 of 10 hemipelvis, and either vesical vein was missing in 3 of them. It was observed that the vesical venous plexus communicated with the deep uterine vein trunk on the medial side of the nerve plane in 6 hemipelvis, while flowed into the deep uterine vein on the lateral side of the nerve plane in 2 hemipelvis, and in the other 2 hemipelvis it directly flowed into the internal iliac vein. (3) It was revealed that autonomic nerves were continuously distributed beneath the ureteral with sagittal plane by HE staining. The average nerve content below the ureteral width was 70.9% of the total in nerve plane by S100 staining. TH and VIP staining showed that the average number of sympathetic fibers was 13.5 and parasympathetic fibers was 8.2, reminding sympathetic predominated.Conclusion:Pelvic autonomic nerves are mainly distributed within the mesangial plane below the ureter, which provides an anatomic justification for NPSRH.
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Objective To compare the clinical effects and the subjective perception of surgeons with three-dimensional (3D) and two-dimensional (2D) laparoscopic C1 radical hysterectomy surgeries for cervical cancer. Methods The retrospective cohort study was conducted. The clinicopathological data of 101 patients with cervical cancer who received C1 laparoscopic radical hysterectomy (C1-LRH) surgery from June 2015 to August 2017 were collected. Of all patients, 42 cases undergoing 3D laparoscopic surgery and 59 cases undergoing 2D laparoscopic surgery were respectively allocated into the C1-3DLRH group or C1-2DLRH group. The clinical effect and the subjective perception of surgeons were compared between the two groups. Results (1) There was no significant difference between the C1-3DLRH group and C1-2DLRH group in terms of age, body mass index (BMI), International Federation of Gynecology and Obstetrics (FIGO) stage, pathologic type, etc. (all P>0.05). Compared with C1-2DLRH group, the operation time was significantly shortened [(192±54) vs (221±54) minutes, P<0.01], blood loss was significantly less [(102±88) vs (167 ± 117) ml, P<0.01], and the success rate of inferior hypogastric plexus (IHP) bladder branch preservation was significantly increased [86% (36/42) vs 66% (39/59), P<0.05] in C1-3DLRH group. There were no significant difference in the number of lymph nodes, the incidence of operative complications, the infection rate, the time of catheterization and the length of hospitalization between the two groups (all P>0.05). The long-term bladder function was evaluated at the twelfth month after operation, 39 patients in the C1-3DLRH group and 53 patients in the C1-2DLRH group were completed the survey. The results showed that 13% (5/39) of the patients in the C1-3DLRH group had long-term bladder dysfunction, which was lower than that 21% (11/53) of the C1-2DLRH group, but there was no significant difference between the two groups (χ2=0.980, P=0.322). (2) A total of 251 laparoscopic surgeons questionnaires were eligible. The incidence of side effects in the first and second generation of 3D and 2D laparoscopic surgeons was 20.4% (10/49), 6.9% (6/87) and 3.5% (4/115), respectively. The incidence of side effects in the first generation of 3D laparoscopic surgeons was higher than that in the second generation of 3D (χ2=5.463, P=0.019) and 2D laparoscopic surgeons (χ2=12.475, P<0.01). There was no difference between the second generation of 3D and 2D laparoscopic surgeons (χ2=1.208, P=0.272). Conclusions 3D laparoscopy is advantageous to the preservation of autonomic nerve in C1-LRH operation and may improve the quality of operation compared with 2D laparoscopy. The second generation of 3D laparoscopic device might overcome the side effects of the surgeons.
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AIM:To investigate the inhibitory effect of ethanol extract of propolis (EEP) on oxidized low-density lipoprotein (ox-LDL)-induced vascular endothelial cell apoptosis and the underlying molecular mechanisms.METHODS:Human umbilical vein endothelial cells (HUVECs) were pretreated with EEP (7.5,15 and 30 mg/L) or 4-phenylbutyric acid (PBA,4 mmol/L) for 1 h and then treated with ox-LDL (100 mg/L) or tunicamycin (TM,4 mg/L) for 24 h.The cell viability and apoptosis were determined by MTT assay and Annexin V-FITC/PI double staining,respectively.The activities of lactic dehydrogenase (LDH) in the medium and caspase-3 in the HUVECs were measured.The protein and mRNA levels of C/EBP homologous protein (CHOP),a proapoptotic molecule under endoplasmic reticulum stress (ERS),and its downstream Bcl-2 were examined by Western blot and real-time PCR,respectively.RESULTS:Like PBA (an ERS inhibitor),EEP protected HUVECs from ox-LDL-induced injury in a dose-dependent manner,as assessed by the increased cell viability and the decreased LDH release,apoptotic rate and caspase-3 activation.The decrease in cell viability and the increases in LDH release,apoptotic rate and caspase-3 activation induced by TM,an ERS inducer,were also attenuated by EEP.Moreover,EEP suppressed ox-LDL-induced CHOP upregulation and Bcl-2 downregulation,and this effect was similar to that of PBA.Similarly,EEP significantly suppressed TM-induced CHOP upregulation both at the protein and mRNA levels.CONCLUSION:EEP may protect HUVECs from ox-LDL-induced apoptosis,and the mechanism is at least partially involved in suppressing CHOP-mediated ERS-associated apoptotic pathway.
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AIM:To investigate the inhibitory effect of allicin on apoptosis and caspase-12 activation of macro-phage-derived foam cells,and to elucidate the underlying molecular mechanisms. METHODS:RAW264.7 macrophages were pretreated with allicin (12.5,25 and 50 mg/L) or 4-phenylbutyric acid(PBA,4 mmol/L) for 1 h and then treated with oxidized low-density lipoprotein(ox-LDL,100 mg/L) or tunicamycin(TM,4 mg/L) for 24 h. The cell viability and apoptosis were examined by MTT assay and flow cytometry with Annexin V-FITC/PI staining,respectively. The activities of caspase-3 in the cells and lactic dehydrogenase (LDH) in the medium were measured. The protein levels of caspase-12 were determined by Western blot. The intracellular lipid accumulation was measured with oil red O staining and the content of intracellular total cholesterol was determined by enzymatic colorimetry. RESULTS:Similar to the endoplasmic reticulum stress (ERS) inhibitor PBA, allicin inhibited ox-LDL-induced injury of RAW264.7 macrophages in a concentration-de-pendent manner,as determined by the increased cell viability and the decreased LDH leakage,apoptosis and caspase-3 ac-tivity. The decrease in cell viability and increases in LDH leakage and apoptosis induced by TM (an ERS inducer) were also suppressed by allicin. Moreover, similar to PBA, allicin remarkably inhibited ox-LDL- or TM-induced activation of caspase-12. Furthermore, allicin remarkably attenuated ox-LDL-induced lipid accumulation in the RAW264.7 cells and foam cells formation in a concentration-dependent manner. CONCLUSION:Allicin may inhibit macrophage-derived foam cell apoptosis induced by ox-LDL,and the mechanism is partially related to suppressing the activation of caspase-12.
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AIM:To investigate the inhibitory effect of ethanol extract of propolis (EEP) on oxidized low-density lipoprotein (ox-LDL)-induced vascular endothelial cell apoptosis and the underlying molecular mechanisms.METHODS:Human umbilical vein endothelial cells (HUVECs) were pretreated with EEP (7.5,15 and 30 mg/L) or 4-phenylbutyric acid (PBA,4 mmol/L) for 1 h and then treated with ox-LDL (100 mg/L) or tunicamycin (TM,4 mg/L) for 24 h.The cell viability and apoptosis were determined by MTT assay and Annexin V-FITC/PI double staining,respectively.The activities of lactic dehydrogenase (LDH) in the medium and caspase-3 in the HUVECs were measured.The protein and mRNA levels of C/EBP homologous protein (CHOP),a proapoptotic molecule under endoplasmic reticulum stress (ERS),and its downstream Bcl-2 were examined by Western blot and real-time PCR,respectively.RESULTS:Like PBA (an ERS inhibitor),EEP protected HUVECs from ox-LDL-induced injury in a dose-dependent manner,as assessed by the increased cell viability and the decreased LDH release,apoptotic rate and caspase-3 activation.The decrease in cell viability and the increases in LDH release,apoptotic rate and caspase-3 activation induced by TM,an ERS inducer,were also attenuated by EEP.Moreover,EEP suppressed ox-LDL-induced CHOP upregulation and Bcl-2 downregulation,and this effect was similar to that of PBA.Similarly,EEP significantly suppressed TM-induced CHOP upregulation both at the protein and mRNA levels.CONCLUSION:EEP may protect HUVECs from ox-LDL-induced apoptosis,and the mechanism is at least partially involved in suppressing CHOP-mediated ERS-associated apoptotic pathway.
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Objective To investigate the expression of elongation of very long-chain fatty acids family member 6 (ELOVL6) in high-grade serous ovarian carcinoma (HSOC), and explore the correlation between its expression and clinical prognosis in these patients. Methods The expression of ELOVL6 at mRNA and protein levels were respectively detected by reverse transcription (RT)-PCR and immune histochemistry method in 12 cases with normal ovarian tissues and 172 cases with HSOC from primary tumor site, forty of which had paired peritoneal metastatic tissues. Results (1) The results tested by RT-PCR showed that ELOVL6 expression in normal ovarian tissue was 4.8±1.1, while 1.2±0.7 in primary tumors and 1.8 ± 0.9 peritoneal metastatic sites in HSOC. Compared with normal ovarian tissue, the level of ELOVL6 mRNA was significantly lower in HSOC (P0.05). The total median survival was 52 months in ELOVL6-positive group and 44 months in ELOVL6-negative group (P>0.05). Conclusion Low expression of ELOVL6 may correlate with the poor differentiation and drug resistance in HSOC.
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<p><b>OBJECTIVE</b>To investigate the clinical characteristics and prognostic factors for patients with stage Ia-IIb cervical cancer with pulmonary metastasis (CCMP).</p><p><b>METHODS</b>Clinical information of 106 patients with stage Ia-IIb cervical cancer with lung metastases admitted in the Chinese Academy of Medical Sciences Cancer Hospital from January 1999 to June 2013 was collected. The efficacy of different therapeutic methods for lung metastases was compared and relevant prognostic factors were analyzed.</p><p><b>RESULTS</b>Pulmonary metastases were detected in 89 (83.9%) out of a total of 106 patients within 2 years after initial treatment of cervical cancer. The median disease-free interval (DFI) of the 106 patients was 13 months. The median survival after CCMP was 18 months, with 2-year and 5-year survival rates of 37.7% and 7.5%, respectively. Metastases were mainly distributed in the inferior lobe of right lung. The surgical treatment was proven to be clinically effective in both unilateral and bilateral CCMP. The response rate was 60.2% in patients choosing non-operative treatment. The univariate analysis showed that pathological type of cervix carcinoma, low degree of differentiation, scope of pulmonary metastasis and whether CCMP simultaneously accompanied with other parts of recurrence were closely related to the prognosis, while the differentiation of cervical tumor was an independent prognostic factor drawn from multivariate regression analysis.</p><p><b>CONCLUSIONS</b>Regular CT lung screening is recommended for patients with stage Ia-IIb cervical cancer during their follow-up period. Both operative and non-operative treatments are feasible therapeutic methods for CCMP. But surgical resection is strictly restricted to certain operative indications. Ia-IIb stage cervical cancer of low differentiation has poorer prognosis after lung metastasis.</p>
Subject(s)
Female , Humans , Carcinoma, Squamous Cell , Lung Neoplasms , Diagnosis , Multivariate Analysis , Neoplasm Recurrence, Local , Neoplasm Staging , Prognosis , Retrospective Studies , Survival Rate , Uterine Cervical Neoplasms , DiagnosisABSTRACT
Objective: To study the chemical constituents of Artemisia anomala. Methods: Various chromatographic techniques were adopted to separate the constituents, and the spectroscopic analysis was used to identify their structures. Results: Seven compounds were isolated and identified as (4aS, 7S, 7aR)-methyl 7-hydroxy-7-methyl-1, 4a, 5, 6, 7, 7a-hexahydrocyclopenta [c] pyran-4- carboxylate (1), rehmaglutin D (2), (E)-6-hydroxy-2, 6-dimethylocta-2, 7-dienoic acid (3), chrysoeriol (4), luteolin (5), apigenin (6), and p-coumaric acid (7). Conclusion: Compound 1 is a new compound named artanoiridoid; Compound 2 is firstly reported in this genus; Compounds 3, 4, and 7 are separated from this plant for the first time.
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G6PDMahidol enzyme is the most common variant in the Achang Chinese ethnic group and clinically manifests as class II. In this study, G6PDMahidol enzyme was characterized by molecular modeling to understand its kinetics. G6PDMahidol, G6PDG487A and G6PDWT proteins were heterologously expressed in the G6PD-deficient DF213 E. coli strain, purified and their steady-state kinetic parameters were determined. Compared with G6PDWT, the Km and Vmax of NADP+ with G6PDG487A were about 28-fold and 12-fold lower, respectively. The Ki values of dehydroepiandrosterone (DHEA), NADPH and ATP with G6PDG487A showed 29.5-fold, 2.36-fold reduction and 1.83-fold increase, respectively. A molecular modeling of G6PDG487A was performed based on the X-ray structure of human G6PD (PDB: 2BH9). It is suggested that Ser-163 might affect the stability of G6PDG487A -helix d and -strand E, besides the conformation of -strand D. In conclusion, the biochemical and structural properties of G6PDG487A and G6PDWT enzymes are significantly different, which may be responsible for clinical diversity of G6PD deficiencies.
Subject(s)
Acute Disease , Adolescent , Anemia, Hemolytic/enzymology , Anemia, Hemolytic/etiology , Asian People , Computer Simulation , Female , Glucosephosphate Dehydrogenase/antagonists & inhibitors , Glucosephosphate Dehydrogenase/chemistry , Glucosephosphate Dehydrogenase/pharmacokinetics , Glucosephosphate Dehydrogenase Deficiency/complications , Glucosephosphate Dehydrogenase Deficiency/enzymology , Humans , Kinetics , Molecular Dynamics Simulation , MutationABSTRACT
The metalloproteinases/disintegrins in the snake venom act as platelet aggregation inhibitor by an antagonism against integrin on platelet through its RGD sequence and may play other important role in cell-cell fusion, cell matrix interaction and other cellular function. Ussurin is a new metalloproteinase/disintegrin that was cloned from Gloydius ussuriensis. Poly (A+) RNA was purified from the total RNA preparation from venom gland of a single G. ussuriensis using the poly (A+) tract-mRNA isolation system. A cDNA library was constructed with the SMART PCR cDNA library construction kit. The cDNA library was screened and the positive clones were selected. The full-length cDNA of Ussurin was obtained. The cDNA encoding the Ussurin precursor has a 51bp 5'-UTR, the open reading frame of Ussurin and a 490 bp 3'-UTR, the open reading frame of Ussurin cDNA nucleotide sequence is 1434 bp and codes for 478 amino acids with a predicted molecular mass of 53.2 kD and an isoelectric point of 5.37. There is no potential N-glycosylation site in the deduced sequence region. Its deduced amino acid sequence consists of four region, a signal sequence of 18 amino acid residues, a zymogen pro-peptide of 171 amino acid residues with a cysteine switch motif (PK-MCGVT) in it, a central metalloproteinase domain of 201 amino acid residues containing a conserved zinc-chelating sequence (HEXXHXXGXXH) and a methionine-turn CIM involving zinc banding also, a space sequence between metalloproteinase domain and disintegrin domain of 15 amino acid residues with a conserved T392, T397, S400, which is specific residues of the P-II snake venom metalloproteinases, a disintegrin domain of 73 amino acid residues with a characteristic RGD region and six-disulfide bonds. Ussurin belongs to P-II class. The cDNA sequence and deduced amino acid sequence of Ussurin precursor were compared with homologous sequence in the GenBank database, the result reveals high degree of homology in sequence and organization pattern of domain with metalloproteinase/disintegrin gene family of other snake species. Compared with the alignment of amino acid sequence of metalloproteinase/disintegrin member, hypervariable regions of this member were revealed, besides they share higher homologous in the zymogen domain. It suggests that the hypervariable regions are the counterparts directly suitable for interacting with different domain of receptors, different receptors or substrates.
Subject(s)
Animals , Amino Acid Sequence , Base Sequence , Cloning, Molecular , Disintegrins , Chemistry , Genetics , Metabolism , Metalloproteases , Chemistry , Genetics , Metabolism , Molecular Sequence Data , Sequence Alignment , Viper Venoms , Genetics , Viperidae , Genetics , MetabolismABSTRACT
[Objective] To explore the mechanism of fever in damp- heat syndrome. [Methods] Hyperlipemia rabbit models were established by feeding fatty - sweet diet. After the injection of endotoxin, the secretion of tumor necrosis factor and interleukin 1 was observed in rabbit model (Group A) and compared with normal rabbits (Group B) . [ Results] The peak of tumor necrosis factor and interleukin 1 secretion was decreased and the decline was slow in Group A. [ Conclusion ] The damp - heat syndrome in seasonal febrile disease is not merely a simple addition of damp and heat, but a result of retention and interaction of the two factors.
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Objective To approach the relationship betwe en glycosaminoglycan metabolism in cartilages and pathogenesis of KBD.Methods Rhesus monkey was fed with grains and water from KBD endemic area for 18 months to produce the animal model with KBD . The glyc osaminoglycans in the monkey cartilage were extracted by the improved Dish method of Bitter. Purified glycosaminoglycans were digested with chondroitinaseABC, and the enzymatic digests were analyzed by HPLC. Results Comparing with those of the control, the glycos aminoglycans in the head of femur, tibia plateau and costal cartilage from the Rhesus monkey fed with grains and water from KBD endemic area were undersulfated . Decreased unsaturated 4-sulphated disaccharide (△Di-4S) from the glycosa minoglycans in the head of femur and tibia plateau and decreased unsaturated 6- sulphated disaccharide (△Di-6S) from the glycosaminoglycans in the costal cart ilage were discovered.Conclusions Detrimental factors in grains and water from KBD endemic area cause undersulfate of the cartilages glycosaminoglycans from Rhesus monkey. The glycosaminoglycans changes have a direct bearing on the patho logical alterations in morphology on the cartilages from the animal model with K BD.