[Hippocampus is involved in 17ß-estradiol exacerbating experimental occlusal inter- ference-induced chronic masseter hyperalgesia in ovariectomized rats].

OBJECTIVE: To investigate the influence of chronic masseter hyperalgesia induced by 17ß-estradiol (E2) and experimental occlusal interference (EOI) on underlying mechanism in hippocampus of ovariectomized (OVX) rats. METHODS: In the study, 32 OVX rats were randomly divided into 4 groups (8 rats/group): The control group was OVX group, and 0 µg/d E2 (vehicle) injection was started 7 d after OVX without EOI; in the experimental group (1) OVX + E2 group, 80 µg/d E2 injection was started 7 d after OVX without EOI; in the experimental group (2) OVX + EOI group, vehicle injection was started 7 d after OVX and EOI was applied 17 d after OVX; in the experimental group (3) OVX + E2 + EOI group, 80 µg/d E2 injection was started 7 d after OVX and EOI was applied 17 d after OVX. Bilateral masseter muscle mechanical withdrawal thresholds were measured before OVX, 7 days after OVX (before E2 injection), 17 days after OVX (10 days after E2 injection and before EOI) and 24 days after OVX (7 days after EOI). Immunofluorescence staining was used to reveal phospho-extracellular signal regulated kinase 1/2 (p-ERK1/2)-positive neurons in CA3 of hippocampus. The protein expression of p-ERK1/2 in hippocampus was detected using Western Blot. RESULTS: Compared with the control group [left side: (135.3±8.5) g, right side: (135.4±10.8) g], bilateral masseter muscle mechanical withdrawal thresholds of OVX+E2 group [left side: (113.3±5.6) g, right side: (112.5 ± 5.6) g] and OVX+EOI group [left side: (93.3±5.4) g, right side: 90.8±5.5) g] were decreased (P < 0.01). Bilateral masseter muscle mechanical withdrawal thresholds were significantly lower in OVX+E2+EOI group [left side: (81.2±6.2) g, right side: 79.8±7.7) g] than in the control, OVX+E2 and OVX+EOI groups (P < 0.05). The proportion of p-ERK1/2 positive neurons in the CA3 region of the hippocampus was increased in the control, OVX+E2, OVX+EOI and OVX+E2+EOI groups in turn, and the difference between the groups was statistically significant (P < 0.05). p-ERK1/2 protein expression was increased in the control, OVX+E2 and OVX+EOI groups in turn, but the difference was not statistically significant (P>0.05). p-ERK1/2 expression was significantly higher in OVX+E2+EOI group than in the other three groups (P < 0.05). CONCLUSION: High concentration of E2 could exacerbated EOI-induced chronic masseter hyperalgesia in ovariectomized rats, and its central mechanism may be related to the upregulation of the phosphorylation of ERK1/2 in hippocampus.

[Application values of right atrial volume, myocardial strain and strain rate in evaluating right atrial function in patients with essential hypertension with normal left ventricular ejection fraction].

Objective: To explore the application values of right atrial (RA) volume, myocardial strain and strain rate in evaluating right atrial functions in patients with essential hypertension (EHT) with normal left ventricular ejection fraction (LVEF). Methods: We retrospectively collected a total of 126 EHT patients with normal LVEF, including 66 males and 60 females, age from 18 to 83(48±15) years from the Affiliated Changzhou No.2 People's Hospital with Nanjing Medical University from March, 2018 to May, 2020. According to the left ventricular mass index, those patients were divided into without left ventricle hypertrophy (LVH) group (n=66) and LVH group (n=60), and 83 healthy volunteers were enrolled as control group, which including 42 males and 41 females, age from 14 to 88 (48±16) years. The total, passive and active RA emptying fraction (total RAEF, passive RAEF and active RAEF) were measured by using RA volumes method. The RA strain (S-reservoir, S-conduit, S-booster pump) and the strain rate (SR-reservoir, SR-conduit, SR-booster pump) were measured by two-dimensional speckle tracking echocardiography. Total RAEF, S-reservoir, and Sr-reservoir represented the reservoir function, while passive RAEF, S-conduit, and Sr-conduit represented the conduit function, and active RAEF, S-booster pump and Sr-booster pump represented the booster pump function, respectively. Than the values between the EHT patients and normal subjects, and the values among the EHT patients without LVH, EHT patients without LVH and normal subjects were compared. Results: The value of active RAEF was significantly lower in EHT patient than normal controls (40.1%±10.5% vs 45.4%±11.1%,P=0.001). There were significant differences among EHT patients without LVH, EHT patients with LVH and normal subjects in passive RAEF and active RAEF (passive RAEF: 40.2%±9.3% vs 36.4%±10.7% vs 36.5%±9.4%, P=0.038, active RAEF: 39.8%±10.6% vs 40.4%±10.4% vs 45.4%±11.1%, P=0.002). There were significant differences in S-reservoir, S-conduit, S-booster pump, SR-reservoir, SR-conduit and SR-booster pump between EHT patients and normal subjects [S-reservoir: 34.8%±13.7% vs 45.0%±13.3%, S-conduit: 18.3%±9.4% vs 26.5%±9.6%, S-booster pump: 16.4%±7.3% vs 18.6%±7.1%, SR-reservoir: (1.6±0.6)/s vs (2.0±0.6)/s, SR-conduit: (-1.2±0.5)/s vs (-1.6±0.6)/s, SR-booster pump: (-1.7±0.6)/s vs (-2.3±0.6)/s; all P<0.05]. The trend tests showed normal controls achieved the largest the absolute values of S-reservoir, S-conduit, SR-reservoir, SR-conduit, SR-booster pump, followed by EHT patients without LVH and EHT patients with LVH(all P<0.001). Conclusions: Using RA volume, myocardial strain and strain rate can detect the RA impaired reservoir, conduit, and the booster pump functions in EHT patients with normal LVEF, and the RA functions were damaged more significantly in EHT patients with LVH.

[Analysis of related factors and prediction of poor ovarian response in patients with controlled ovarian stimulation].

Objective: To explore the related factors of poor ovarian response (POR) in patients receiving controlled ovarian stimulation (COS) and to establish the nomogram for predicting POR in patients who received in vitro fertilization or intracytoplasmic sperm injection (IVF/ICSI). Methods: In this retrospective research, clinical data of 17 164 cycles of patients who received IVF/ICSI treatment at Henan Provincial People's Hospital from September 1st, 2016 to September 1st, 2020 were analyzed. Independent correlative factors affecting the occurrence of POR were screened by logistic regression, which were the model enrollment variables in the prediction model. Totally 13 266 cycles with well-record of enrollment variables were screened, and these data were randomly divided into model group (9 896 patients) and validation group (3 370 patients) according to 3∶1. The nomogram was established according to the regression coefficient of the relevant variables. The prediction accuracy of the nomogram was evaluated by calculating area under the receiver operating characteristic curve (AUC). Results: Multivariate logistic regression analysis showed age, infertility type, body mass index, anti-Müllerian hormone, basal follicle stimulating hormone, basal estrogen, antral follicle number, previous times of POR, history of ovarian surgery, ovulation stimulation protocol and average amount of gonadotropin were independent correlative factors affecting the occurrence of POR (all P<0.05). In the model group, according to the above factors, the prediction model and nomogram of POR risk were constructed and the validation group verified the model. The AUC of the model group was 0.893 (95%CI: 0.885-0.900), and the AUC of the validation group was 0.890 (95%CI: 0.878-0.903). Conclusion: The influencing factors of POR after COS in patients treated by IVF/ICSI are screened, and the nomogram for predicting POR established in this study is proved to be effective, simple, intuitive and clear in predicting the occurrence of POR.

[Clinical, pathological and genetic characteristics of 8 patients with distal hereditary motor neuropathy].

OBJECTIVE: Distal hereditary motor neuropathy (dHMN) comprises a heterogeneous group of inherited disorders associated with neurodegeneration of motor nerves and neurons, mainly charac-terized by progressive atrophy and weakness of distal muscle without clinical or electrophysiological sensory abnormalities. To improve the recognition and diagnosis of the disease, we summarized the clinical manifestations, electrophysiological, pathological, and genetic characteristics in eight patients with dHMN. METHODS: Eight probands from different families diagnosed with dHMN were recruited in this study between June 2018 and April 2019 at Peking University People's Hospital. Eight patients underwent complete neurological examination and standard electrophysiological examinations. The clinical criteria were consistent with the patients presenting with a pure motor neuropathy with no sensory changes on electrophysiology. The detailed clinical symptoms, neurophysiological examinations, pathological features and gene mutations were analyzed retrospectively. Genetic testing was performed on the eight patients using targeted next-generation sequencing panel for inherited neuromuscular disorder and was combined with segregation analysis. RESULTS: The age of onset ranged between 11 and 64 years (median 39.5 years) in our dHMN patients. All the cases showed a slowly progressive disease course, mainly characterized by distal limb muscle weakness and atrophy. The motor nerve conduction revealed decreased compound muscle action potential amplitude and velocity, while the sensory nerve conduction velocities and action potentials were not affected. Needle electromyography indicated neurogenic chronic denervation in all patients. Muscle biopsy performed in two patients demonstrated neurogenic skeletal muscle damage. Sural nerve biopsy was performed in one patient, Semithin sections shows relatively normal density and structure of large myelinated fibers, except very few fibers with thin myelin sheaths, which suggested very mild sensory nerve involvement. Eight different genes known to be associated with dHMN were identified in the patients by next-generation sequencing, pathogenic dHMN mutations were identified in three genes, and the detection rate of confirmed genetic diagnosis of dHMN was 37.5% (3/8). Whereas five variants of uncertain significance (VUS) were identified, among which two novel variants co-segregated the phenotype. CONCLUSION: dHMN is a group of inherited peripheral neuropathies with great clinical and genetic heterogeneity. Next-generation sequencing is widely used to discover pathogenic genes in patients with dHMN, but more than half of the patients still remain genetically unknown.

[Influencing factors of thrombosis besides antiphospholipid antibodies in patients with antiphospholipid syndrome].

Objective: To identify the influencing factors of thrombosis besides antiphospholipid antibodies in patients with antiphospholipid syndrome (APS). Methods: The 169 patients diagnosed with APS were enrolled according to the current APS classification criteria from January 2003 to August 2017 in Peking University People's Hospital. There were 23 males and 146 females with a mean age of (41±15) years. Antiphospholipid antibodies, including anticardiolipin (aCL), anti-ß2glycoprotein-1 (ß2GP1) antibodies and antibodies to the phosphatidylserine-prothrombin complex (aPS/PT), were determined by enzyme-linked immunosorbent assay (ELISA) methods. Lupus anticoagulant (LA) was identified using the STA Compact coagulation testing system. The differences of clinical and laboratory characteristics between patients with and without thrombosis were analyzed (100 cases and 69 cases, respectively). The influencing factors for thrombosis in patients with APS were determined using binary logistic regression. Results: Compared with patients without thrombosis, patients with thrombosis were older and had a longer disease duration ((45±17) years vs (35±9) years and M(Q1, Q3) 12.0(3.8, 84.0) months vs 48.0(12.0, 108.0) months, both P<0.05). The percentage of male, primary APS, smoking, low blood platelet count, hypertension, and diabetes in patients with thrombosis were significantly higher than those in patients without thrombosis (all P<0.05). Similarly, the rates of antinuclear antibodies positive, aCL positive, aPS/PT-IgM positive, and aPS/PT-IgG positive in patients with thrombosis were significantly higher than those in patients without thrombosis (all P<0.05). The levels of D-dimer in patients with thrombosis were significantly higher than that in patients without thrombosis (P<0.05). There was significant difference in global anti-phospholipid syndrome score (GAPSS) between patients with and without thrombosis (P<0.05). The GAPSS score was also significantly higher in patients with arterial thrombosis than that in patients with venous thrombosis (P<0.05). Smoking and D-dimer levels were independent influencing factors for thrombosis in patients with APS (smoking: OR=11.222, 95%CI:1.119-112.544, P=0.040, D-dimer levels: OR=1.002, 95%CI: 1.000-1.003, P=0.037). Conclusions: Thrombotic APS patients are older and have a longer suffering duration, a higher ratio of male, primary APS, smoking, hypertension, lower blood platelet count, diabetes, higher GAPSS scale, and higher D-dimer levels. Smoking and D-Dimer levels may be independent risk factors for thrombosis in patients with APS.

[Analysis of the level of T lymphocyte subsets and the relationship with the prognosis of pulmonary fibrosis in patients with paraquat poisoning].

Objective: To analyze the levels of T lymphocyte subsets (CD3, CD4, CD8 and CD4/CD8) in patients with paraquat poisoning, and to explore the relationship between the changes of T lymphocyte subsets and the prognosis of pulmonary fibrosis. Methods: In October 2019, a total of 47 patients with oral 20% paraquat low water solvent poisoning in Guangzhou 12th people's Hospital from June 2018 to June 2019 were selected as the research objects. Patients were divided into early death group (16 cases died within 2 weeks) and non early death group (31 cases survived more than 2 weeks) . The non early death group was divided into pulmonary fibrosis group (23 cases) and normal lung group (8 cases) . 20 healthy people in the same period were randomly selected as the control group. The neutrophils (N) , C reaction protein (CRP) , alanine aminotransferase (ALT) , creatinine (Cr) , amylase (aAMY) , creatine kinase isoenzyme (CKMB) , pH, HCO(3)(-), blood oxygen saturation (SO(2)) and lactic acid (Lac) of patients poisoned within 3 d were examined every day. Independent sample t-test was used for inter group comparison, and paired sample t-test was used for intra group comparison. Results: Compared with non early death group, the levels of N, CRP, ALT, Cr, aAMY, CKMB and Lac in early death group increased (P<0.05) , while pH and HCO(3)(-) decreased (P<0.05) . Compared with the control group, the levels of CD3, CD4 and CD4/CD8 were decreased on the first day in the early death group and non early death group (P<0.05) , and the levels of CD3, CD4 and CD4/CD8 were decreased on the 15th day in the pulmonary fibrosis group (P<0.05) . Compared with the normal lung group, the levels of CD3, CD4 and CD4/CD8 in the pulmonary fibrosis group decreased on the 15th day (P<0.05) . Conclusion: The persistent low cellular immune function in patients with paraquat poisoning is related to the progress of pulmonary fibrosis, which is an important factor affecting the prognosis of patients with pulmonary fibrosis.

[Co-existence of Guillain-Barré syndrome and Behcet syndrome: A case report].

A 40-year-old male patient was referred to our department with complains of recurrent oral ulcer for more than 20 years and vulvar ulcer for more than 10 years. He presented with a 3-month history of right external ophthalmoplegia. More than 10 days ago, the patient received ganglioside infusion. And one week ago, he developed numbness and pain of his lambs, and progressive myasthenia, accompanied by right blepharoptosis and dysuria. On exam, motor strength was graded 0/5 in the lower and the upper extremities. Deep tendon reflexes were diminished in extremities. His admission medical examination: hemoglobin (HGB), white cell and platelet counts were normal. C-reactive protein (CRP) was negative. Erythrocyte sedimentation rate (ESR) 53 mm/h. Antinuclear antibody (ANA), anti-dsDNA antibody, anti-Smith antibody, anti-cardiolipin antibody and human leucocyte antigen B51 were all within normal range. The etiological tests of influenza A pathogen, influenza B pathogen, parainfluenza virus, enterovirus and parvovirus were all negative. He tested positive for serum anti-GM1 IgG. Cerebrospinal fluid had a normal white cell count, an elevated protein content. Gram staining, culture and PCR detection for varicella-zoster virus, cytomegalovirus and herpes simplex virus were all negative. Antibodies associated with autoimmune encephalitis and paraneoplastic syndrome were negative in cerebrospinal fluid. Electromyography and nerve conduction studies showed a severe axonal damage affecting motor nerves. No obvious abnormalities were observed in his magnetic resonance imaging of brain and cavernous sinus. The patient was diagnosed with Behcet syndrome complicated with acute Guillain-Barré syndrome. He received intravenous methylprednisolone, intravenous immunoglobulin (IVIg) therapy, plasma exchange and rituximab treatment. After treatment, the patient's muscle strength of limbs was restored to grade 1, blepharoptosis and pain disappeared. The nervous system involvement of Behcet syndrome is relatively rare, especially combined with Guillain-Barré syndrome, which is easy to cause misdiagnosis. The treatment of Behcet syndrome complicated with acute Guillain-Barré syndrome includes the treatment of primary disease, plasma exchange and IVIg therapy. In addition, supportive treatment is very important for such patients. The focus of treatment is to avoid respiratory insufficiency, prevent deep vein thrombosis, monitor cardiac function and hemodynamics. Pain-relieving, physical exercise and psychological support are often under-recognized. The rehabilitation treatment is very important to improve the prognosis and quality of life of patients. What we need to learn is that when the symptoms and signs of the nervous system are difficult to be explained by neuro-Behcet syndrome alone, we should be alert to the possibility of other nervous system diseases.

Function of IL-33 in Wound Age Estimation of Skin Wounds in Mice.

ABSTRACT: Objective To explore the application value of interleukin-33 （IL-33） in wound age estimation in forensic practice by observing the sequential changes of IL-33 after skin wound. Methods Skin wound models were generated on the back of mice with a round file of 5 mm in diameter. Skin samples of the same size were taken from the same parts of mice in control group and injury group 1 h, 3 h, 6 h, 12 h, 1 d, 3 d, 5 d, 7 d and 10 d after skin wound. Hematoxylin-eosin （HE） staining method was applied to observe the morphological changes in the recovering process after skin wound. Western blotting, immunohistochemistry staining and double immunofluorescence staining methods were applied to detect the expression changes of IL-33 in the skin wound samples. Results The results of Western blotting showed that the expression of IL-33 protein decreased slightly at 3 h after skin wound, increased gradually at 6 h after skin wound, and reached the peak value at 3 d, then decreased gradually. Immunohistochemistry staining results showed that faint positive expression of IL-33 was observed in epidermis, hair follicles, sebaceous glands and dermal resident cells of the control group skin. The positive cell rate of IL-33 increased at 3 h after skin wound and reached the peak value at 3 d, then decreased gradually. The results of double immunofluorescence staining showed that the majority of IL-33 positive cells from 1 d to 3 d after wound were macrophages, while the majority of IL-33 positive cells from 5 d to 7 d after wound were myofibroblasts. In addition, the results of HE staining showed that the wound healing process of the skin wound model was consistent with the pathological development law of inflammation. Conclusion IL-33 could become a reference index for wound age estimation of skin wound in forensic practice.

Application of Virtual Anatomy Technology in Postmortem Examination of Medical Dispute Cases.

ABSTRACT: Objective To discuss the application value of CT scanning technology in cause of death determination of medical dispute cases. Methods From July 2017 to December 2018, postmortem CT imaging data of 12 medical dispute cases were collected. CT imaging diagnosis results and anatomy findings as well as differences between antemortem and postmortem CT diagnosis were compared. The advantages and disadvantages of CT routine tests of the cadavers in terms of the diagnosis of disease and damage were analyzed. Results The comparison between CT imaging diagnosis and anatomical findings showed that CT scans had advantages in the diagnosis of disease and damage with large differences in density changes, such as atelectasis, pneumonia, calcification, fracture and hemorrhage, etc. The comparison of CT diagnosis in antemortem and postmortem examination showed that the cadavers of medical dispute cases were well preserved and that postmortem CT scan was meaningful for the diagnosis of antemortem diseases. Conclusion Virtual anatomy technology has a relatively high application value in postmortem examination of medical dispute cases. It can provide effective information for the appraisers before the autopsy and can also provide a reference for cause of death analysis when the anatomy cannot be performed.

[Detection of neuron-specific enolase in patients with subacute 1, 2-dichloroethane poisoning].

Objective: To investigate the changes of neuron-specific enolase (NSE) in serum and cerebrospinal fluid of patients with subacute 1, 2-dichloroethane (DCE) poisoning. Methods: Ten patients with subacute 1, 2-DCE poisoning hospitalized in Guangzhou 12th Municipal People's Hospital from December 2014 to March 2019 were taken as the subacute 1, 2-DCE poisoning group, 34 typical acute toxic encephalopathy patients hospitalized at the same time as typical acute toxic encephalopathy group, 40 healthy physical examinees as normal control group. The levels of serum NSE in patients of subacute 1, 2-DCE poisoning and typical acute toxic encephalopathy group during onset and improvement were detected by chemiluminescence method, and the results were analyzed statistically. The level of NSE in cerebrospinal fluid of subacute 1, 2-DCE poisoning group was detected and analyzed its correlation with the level of NSE in serum. Using receiver operator characteristic (ROC) curve to analyze the diagnostic efficacy of NSE in subacute 1, 2-DCE poisoning and typical acute toxic encephalopathy (area under curve, AUC) . Results: There was no significant difference between the serum NSE level of the patients with subacute 1, 2-DCE poisoning in the onset group and the normal control group and the improvement group (P>0.05) . The serum NSE level of subacute 1, 2-DCE poisoning in the improvement group was lower than those in the normal control group (P<0.01) . The serum NSE level of the subacute 1, 2-DCE poisoning in the onset group was lower than those in the typical acute toxic encephalopathy in the onset group (P<0.01) . There was no linear correlation between cerebrospinal fluid NSE and serum NSE in patients with subacute 1, 2-DCE poisoning (r=-0.183, P=0.52) . ROC curve showed that the AUC of serum NSE in diagnosing subacute 1, 2-DCE poisoning and typical acute toxic encephalopathy were 0.661 and 0.726, respectively. Conclusion: There is no significant change in serum NSE in patients with subacute 1, 2-DCE poisoning.

Expression of Annexin A1 during Skin Incised Wound Healing in Mice.

OBJECTIVES: To investigate the expression changes of annexin A1 （ANXA1） during the process of skin incision healing, and to explore its expression and function during skin injury repair. METHODS: The skin injury model of mice was prepared, and skin tissues of the controls and the injured group at 6 h, 12 h, 1 d, 3 d, 5 d, 7 d, 10 d and 14 d after injuries were taken. The morphological changes of the wound were observed by hematoxylin-eosin （HE） staining, and the expression of ANXA1 was detected by immunohistochemistry （IHC） and Western blotting. RESULTS: HE staining showed normal healing of skin wounds. IHC results revealed that ANXA1 was expressed in the epidermis, hair follicle, sebaceous gland and vascular endothelium. In the injured group, the expression of ANXA1 was enhanced in epidermis and skin appendages around the wound 6-12 h after injury, and ANXA1 was also highly expressed in neutrophils and a small number of mononuclear cells. ANXA1 was mainly positively expressed in monocytes, neovascular endothelial cells and fibroblasts, and small amount of fibroblasts at 1-3 d, 5-10 d, and 14 d after injury, respectively. Western blotting showed that, compared with the controls, the expression of ANXA1 was significantly increased at 6 h after injury, peaked at 1 d, and then decreased gradually in the injured group. CONCLUSIONS: ANXA1 may be involved in the regulation of skin damage repair, with time-dependent expression during skin wound healing, and thus is expected to be a biological marker for inferring the wound formation time.

Time-dependent Expression and Distribution of AChE during the Skin Incised Wound Healing in Mice.

ABSTRACT: Objective To study the time-dependent expression and distribution of acetylcholinesterase （AChE） during skin incised wound healing in mice, and discuss its effect in wound healing as well as the feasibility of using it as a reference index for wound age estimation. Methods A total of 45 C57BL/KsJ mice were randomly divided into one control group and eight incised groups. The skin incised wound model was established in the incised groups with samples of skin wounds taken at 6 h, 12 h, 1 d, 3 d, 5 d, 7 d, 10 d and 14 d post-injury respectively, while the uninjured skin tissue was extracted in the control group. Expression and distribution of AChE in skin samples were detected by immunohistochemistry, double immunofluorescence and Western blotting. Results Immunohistochemistry results indicated that AChE was mainly detected in infiltrating polymorphonuclear cells （PMNs） 6 to 12 h post-injury. A large number of AChE-positive mononuclear cells （MNCs） were observed 1 to 3 d post-injury. The AChE-positive cells were mainly fibroblastic cells （FBCs） 5 to 14 d post-injury. The ratio of the AChE-positive cells increased initially 6 h post-injury, and reached the peak at 1 d post-injury. Double immunofluorescent staining showed that the majority of AChE-positive MNCs and FBCs expressed macrophage marker and myofibroblast marker, respectively. Western blotting results showed that the relative expression level of AChE in the incised group was higher than that in the control group averagely, reached the peak at 1 d post-injury, then reached a second peak at 7 d post-injury. Conclusion The expression of AChE is found in PMNs, macrophages and myofibroblast during skin wound healing, which indicates it might be involved in the adjustment of inflammatory response and fibrotic repair after injury. Moreover, combined use of various methods for the detection of the expression of AChE would provide reference for skin wound age estimation.

Relationship between the Number of Neutrophils and Myofibroblasts during Diabetic Wound Healing and Wound Age.

ABSTRACT: Objective To investigate the sequential changes of the number of neutrophils and myofibroblasts during diabetic wound healing, and discuss its application value in wound age estimation. Methods Diabetic DB mice and mice of the same age in the normal control group were selected, a wound healing model was established, wound samples were taken at different time points, while the number of neutrophils and myofibroblasts during diabetic wound healing were determined by immunohistochemical staining technique. Results The number of infiltrated neutrophils in the wounds of control and diabetic groups reached the peak respectively at 12 h and 5 d after injury. Compared with the control group, the number of neutrophils in the diabetic group decreased significantly from 6 h to 1 d after injury, but increased markedly from 5 d to 14 d. From 5 d to 10 d after injury, the average number of neutrophils at high magnification in wounds of the diabetic group was over 30, while that of neutrophils in wounds of the control group was less than 20. Myofibroblasts appeared in wounds from 3 d to 14 d after injury in the control group and from 5 d to 14 d after injury in the diabetic group. The difference in the number of myofibroblasts in wounds between control group and diabetic group from 3 to 7 d after injury had statistical significance. Conclusion In comparison with normal wound healing, the number of neutrophils and myofibroblasts during diabetic wound healing shows different sequential changes. The results of this study can provide reference for wound age estimation of patients with severe diabetes.

[Study on purification effect of formaldehyde in cleanroom by new return air device].

Objective: To effectively reduce the concentration of poisons in cleanroom, protect the health of workers, realize the optimization and automatic control of the new return air device. And the influence of initial concentration, air volume, temperature and relative humidity of formaldehyde on the purification effect of the new return air device was explored. Methods: The purification effect of the new return air device installed with the activated carbon and the photocatalyst purification net or ordinary activated carbon purification network was tested in a 60 m(3) simulated cleanroom. The concentration of formaldehyde was determined by solution absorption-phenol reagent spectrophotometry. Based on the single factor experiment to determine the combination of two purification nets. The effects of air volume, initial formaldehyde concentration, temperature and relative humidity on the purification effect of the new return air device were investigated by orthogonal test. Then, the performance parameters of the return air device to purify formaldehyde were determined. Results: The formaldehyde purification efficiency of the two types of purification nets in the new return air device was higher than that of the ordinary activated carbon purification network (P<0.05) . The combination of activated carbon and photocatalyst purification net has no effect on the formaldehyde purification efficiency of the return air device (P>0.05) . According to the direct analysis and variance analysis, air volume was the most sensitive factor (F value is 18.894, P<0.05) , followed by initial concentration (F value is 16.128, P<0.05) , while temperature and relative humidity have little effect (F value is 0.041 and 0.599, respectively, P>0.05) . LSD analysis showed that there was no significant difference in the purification efficiency of formaldehyde between 475 m(3)/h and 626 m(3)/h (P>0.05) . From the perspective of formaldehyde purification efficiency and energy saving, when the air volume is set to 475 m(3)/h, the new return air device has higher purification efficiency for high concentration of formaldehyde. Conclusion: The new return air device consisting of activated carbon and photocatalyst purification net can play a good purification role in cleanroom with different temperatures and different humidity. Its formaldehyde purification efficiency is affected by air volume and initial concentration.

[Curative effect analysis of bile reinfusion combined with enteral nutrition support before surgery of hilar cholangiocarcinoma].

Objective: To investigate the clinical effect of bile reinfusion combined with enteral nutrition support before surgery for hilar cholangiocarcinoma. Methods: A retrospective analysis of patients with hilar cholangiocarcinoma who underwent surgical treatment at Nanjing Drum Tower Hospital Hepato-biliary-pancreatic Surgery Department from July 2010 to August 2017 was completed.A total of 52 cases were finally enrolled in our study.All the patients included, on the basis of whether they received preoperative drainage and bile reinfusion, were divided into non-drainage group(n=15) and drainage group(n=37). Differences of clinical indicators, including operation time, intraoperative bleeding and serum liver function index levels at day 1, 3, 7 postoperative, postoperative complications(liver failure, biliary fistula, pleural effusion, peritoneal effusion, abdominal cavity infection, death in hospital), tumor classification, R0 resection, postoperative hospitalization time between the 2 groups were analyzed. At the same time, in the drainage group, patients were divided into non-enteral nutrition subgroup(n=13) and enteral nutrition subgroup(n=24) according to whether they received enteral nutrition before operation. The normal distribution data of the group was statistically analyzed by independent sample t test, the non-normal distribution data of the group was statistically analyzed by rank-sum test. The count data was statistically analyzed by non-calibration and correction of the square test. Results: There was no statistically significant difference in general infomation such as age, gender, and serum liver function between non-drainage group and drainage group(P>0.05). There was no statistically significant difference in general information such as age, gender, and serum liver function between non-enteral nutrition group and enteral nutrition group(P>0.05). The rate of vascular resection and reconstruction(33.3%) and operating time(10.8(2.2)h) in drainage group were both higher than those in non-drainage group(6.7% and 8.3(3.0)h), the differences were both statistically significant(χ(2)=4.397, Z=1.595; both P<0.05). The level of AST at the 7th day after surgery in drainage group(32.8(17.3)U/L) was significantly lower than that in non-drainage group(55.0(64.7)U/L), the difference was statistically significant(Z=-2.212, P<0.05). The level of TBil at 1st day after surgery in drainage group(43.6(91.2)µmol/L) was lower than that in non-drainage group(91.2(188.4)µmol/L), the difference was statistically significant(Z=-2.150, P<0.05). The rate of pancreatoduodenectomy(25.0%) and average operating time(11.1(1.3)h) in the enteral nutrition group were both higher than those in the non-enteral nutrition group(0, 9.0(2.6)h). The differences were both statistically significant(χ(2)=3.879, Z=-2.693; P<0.05). The average level of AST at the 1st day after surgery in enteral nutrition group(396.4(268.3)U/L) was significantly lower than that in non-enteral nutrition group(642.5(341.1)U/L), the difference was statistically significant(Z=-2.483, P<0.05). The average level of TBil at the 1st, 3th day after surgery in enteral nutrition group(38.8(21.5)µmol/L and 30.0(25.6)µmol/L) were both lower than those in non-enteral nutrition group(60.9(75.2)µmol/L and 46.5(50.0)µmol/L), the differences were both statistically significant(Z=-2.416, -2.026; P<0.05). The level of CRP at 1st, 3th day after surgery((41.9±31.1)mg/L, (50.8±31.4)mg/L)in enteral nutrition subgroup was lower than that in non-enteral nutrition subgroup((64.4±33.6)mg/L, (74.1±35.3)mg/L), the differences were both statistically significant(t=1.456, 1.675; P<0.05). Conclusion: Based on the present study , there is no effective improvement on postoperative recovery using bile reinfusion combined with nutrition support before R0 resection of hilar cholangiocarcinoma.

[Relationship between FoxO1 Expression and Wound Age during Skin Incised Wound Healing].

OBJECTIVES: To investigate FoxO1 expression and its time-dependent changes during the skin incised wound healing. METHODS: After the establishment of the skin incised wound model in mice, the FoxO1 expression of skin in different time periods was detected by immunohistochemistry and Western blotting. RESULTS: Immunohistochemistry staining showed that FoxO1 was weakly expressed in a few fibroblasts of epidermis, hair follicles, sebaceous glands, vessel endothelium and dermis in the control group. The FoxO1 expression was enhanced in the epidermis and skin appendages around the wound during 6-12 h after injury, which could be detected in the infiltrating neutrophils and a small number of monocytes. FoxO1 was mainly expressed in monocytes during 1-3 d after injury, and in neovascular endothelial cells and fibroblasts during 5-10 d. On the 14th day after injury, the FoxO1 expression still could be detected in a few fibroblasts. The Western blotting results showed that the FoxO1 expression quantity of the tissue samples in injury group was higher than in control group. The FoxO1 expression peaked at 12 h and 7 d after injury. CONCLUSIONS: FoxO1 is time-dependently expressed in skin wound healing, which can be a useful marker for wound age determination.

Effects of forkhead box C2 on carcinogenesis and lymphatic metastasis in endometrial carcinoma.

We investigated the distribution of endometrial lymphatic vessels and expression of forkhead box C2 (FOXC2) in normal endometrium during menstrual cycle and in endometrial adenocarcinoma. Full-thickness uterine samples and endometrial adenocarcinoma samples were collected for immunohistochemical analysis using D2-40 and FOXC2 mouse monoclonal antibodies. The lymphatic vessel density (LVD) of the endometrium was significantly reduced compared with the myometrium during the cycle. Intra-tumoral LVD was significantly decreased in both stages of endometrioid adenocarcinoma compared with normal endometrium and myometrium. Intra-tumoral LVD significantly decreased from stage IA to stage IIIC. Peri-tumoral LVD for stage IA and stage IIIC tumors was significantly increased compared with normal endometrial LVD, but decreased compared with normal myometrial LVD. Stage IIIC showed increased peri-tumoral LVD when compared with stage IA. The positive rate of FOXC2 was 73.3% in proliferative endometrium and 80% in secretory endometrium. Secretory endometrium showed significantly increased FOXC2 expression compared with proliferative endometrium. Endometrioid adenocarcinoma showed significantly increased FOXC2 expression compared with normal endometrium, both in the epithelium and stroma. FOXC2 expression in the stroma significantly increased when pelvic and/or para-aotic lymph nodes were involved. FOXC2 was immunolocalized in low-risk endometrial carcinoma in endometrioid adenocarcinoma, but not in normal endometrium. Endometrial lymphatic vessels were located in normal endometrium and myometrium across the menstrual cycle and in intra-and peri-endometrioid adenocarcinoma, and increased in endometrial adenocarcinoma. Peri-tumoral lymphatics were associated with increased lymphatic metastasis. FOXC2 may be associated with the genesis of endometrial carcinoma and lymphangiogensis in endometrial adenocarcinoma in intra- and peri-tumoral lymphatics.

Genome-wide polymorphisms between the parents of an elite hybrid rice and the development of a novel set of PCR-based InDel markers.

Genome-wide re-sequencing of the Zhenshan 97 (ZS97) and Milyang 46 (MY46) parents of an elite three-line hybrid rice developed in China resulted in the generation of 9.91 G bases of data with an effective sequencing depth of 11.66x and 11.51x, respectively. Detection of genome-wide DNA polymorphisms, single nucleotide polymorphisms (SNPs), short insertions/deletions (InDels; 1-5 bp), and structural variations (SVs), which is an invaluable variation resource for genetic research and molecular marker-assisted breeding, was conducted by comparing whole-genome re-sequencing data. A total of 364,488 SNPs, 61,181 InDels and 6298 SVs were detected in ZS97 and 364,179 SNPs, 61,984 InDels and 6408 SVs were detected in MY46 compared to the 9311 reference sequence. Synteny analysis of the variation revealed a total of 77,013 identical and 181,737 different SNPs and 15,021 identical and 1205 different InDels between ZS97 and MY46, respectively. A total of 180 InDels 3-8 bp in length between ZS97 and MY46 were selected for experimental validation; 160 polymerase chain reaction products were efficiently separated on 6% non-denaturing polyacrylamide gels. Identification of genome-wide variation among the parents of the elite hybrid as well as the set of 160 polymerase chain reaction-based InDel markers will facilitate future genetic studies and the molecular breeding of hybrid rice.

The presence of advanced lesions and associating risk factors for advanced cervical carcinoma in patients with atypical sguamous cells of undetermined significance.

PURPOSE OF INVESTIGATION: To characterize histopathological status, high-risk human papillomavirus (hr-HPV) infection status, and associated risk factors in patients with atypical squamous cells of undetermined significance (ASCUS). MATERIALS AND METHODS: Cervical biopsies obtained from 130 ASCUS patients were subjected to histopathological examination and hr-HPV testing. Associations between advanced lesions and hr-HPV load or age were analyzed, and the confounding factors for high-grade cervical lesions were identified. RESULTS: Cervical biopsies from ASCUS patients had a wide range of pathological states, ranging from normal to invasive cervical carcinoma. High-risk HPV infection was significantly associated with advanced cervical lesions in ASCUS patients; hr-HPV infection and the number of gestations were risk factors for developing advanced cervical disease. CONCLUSIONS: A significant portion of ASCUS patients harbor advanced cervical lesions. The number of gestations and hr-HPV infection can increase the risk of developing advanced cervical lesions in ASCUS patients.