Volume 87  ·   Issue 25  ·  2007

Objective to investigate the clinical significance of atypical squamous epithelial cells (ASCUS) and low-grade squamous intraepithelial lesions (LSIL) in cervical cytology. Methods 405 cases of ASCUS and 395 cases of LSIL were examined by colposcopy, and biopsy of cervix and/or vulva, endometrial curettage of cervix (ECC) and electrothermal circle resection of cervix (LEEP) were performed under their guidance. The sensitivity, specificity, positive predictive value and coincidence rate of colposcopy in the diagnosis of cervical cancer and its precancerous lesions were evaluated with histopathological diagnosis as the gold standard. Results (1) Histopathological diagnosis results: 231 cases (57.04%) of 405 cases of ASCUS were chronic cervicitis, 141 cases (34.81%) of low-grade squamous intraepithelial lesions (LSIL), 16 cases (3.95%) of high-grade squamous intraepithelial lesions (HSIL), and 2 cases (0.49%) of stage Ⅰ a1 cervical squamous carcinoma (SCC); There were 15 cases (3.70%) of vulvar intraepithelial neoplasia (VIN), 135 cases (34.18%) of 395 cases of LSIL were chronic cervicitis, 201 cases (50.89%) of cervical LSIL, 41 cases (10.38%) of HSIL, 2 cases (0.51%) of SCC Ⅰ a1 stage; (2) The sensitivity, specificity, positive predictive value and histological coincidence rate of colposcopy in the diagnosis of cervical cancer and its precancerous lesions were 96.17%, 58.41%, 73.34% and 79.63%, respectively. (3) Follow-up results: All patients with HSIL and HSIL + were visited, and the lesions regressed after clinical intervention, and there was no progression after follow-up for more than 1 year. All 738 patients with cervicitis, LSIL and VIN were visited within half a year, and 501 of them were followed up for more than 1 year (follow-up rate 67.89%). Conclusion The histological classification of ASCUS/LSIL patients in cervical cytology is varied, including benign changes of cervical epithelium, different grades of CIN and a few cancers.

Objective to investigate the expression and significance of Survivin gene in thyroid carcinoma. Methods the expression of Survivin gene in 98 thyroid carcinoma and its adjacent tissues was detected by Western blotting and reverse transcription-polymerase chain reaction (RT-PCR). The apoptosis index (AI) of thyroid carcinoma cells was detected by in situ end labeling (TUNEL). Results the expression of Survivin protein in thyroid carcinoma tissues was consistent with the results of RT-PCR. 68.37% of thyroid carcinoma tissues expressed Survivin protein and mRNA, but none in adjacent tissues had positive expression. The positive expression rate was closely related to pathological classification, lymph node metastasis and hematogenous metastasis. Apoptotic cells could be detected in all thyroid carcinoma tissues. The average AI of Survivin expression positive group (0.48±0.38) was significantly lower than that of negative group (2.98±0.12, P =0.0082). Conclusion the abnormal expression of Survivin gene in thyroid carcinoma tissues; Its expression is closely related to the degree of differentiation, lymph node metastasis and hematogenous metastasis; The inhibition of apoptosis by Survivin may be a key factor involved in tumorigenesis, development and outcome, and Survivin gene will become a new target of gene therapy for thyroid cancer.

Objective to summarize the indication selection and treatment experience of non-stop coronary artery bypass grafting and pulmonary lobectomy in patients with pulmonary tumor complicated with coronary heart disease. Methods 10 patients with pulmonary tumor complicated with coronary heart disease underwent non-stop coronary artery bypass grafting and pulmonary lobectomy at the same time since 2000. Preoperative cTNM stages of pulmonary tumor were Ⅰ B in 4 cases, ⅡA in 2 cases and ⅡB in 4 cases. Coronary angiography showed left main disease in 2 cases, three vessel disease in 7 cases and single lesion in 1 case. Intraoperative anastomosis of internal mammary artery and left anterior descending artery and great saphenous vein bypass grafting were completed. Lobectomy plus lymph node dissection were performed in 8 cases, and partial lung resection was performed in 2 cases. The index of preoperative cardiopulmonary function evaluation and the basis of selection of surgical indication were analyzed. The key points of intraoperative operation and perioperative management were summarized. Results 10 patients had a smooth operation, passed the perioperative period smoothly, and their cardiopulmonary function recovered well. All of them were cured and discharged. There were 2 cases of inflammatory pseudotumor, 1 case of pTNM stage Ia, 4 cases of stage IIa, and 3 cases of stage IIb. Conclusion Concurrent lobectomy with non-stop coronary artery bypass grafting should be carried out with strict control of surgical indications, correct control of intraoperative and postoperative management principles, and it is safe and feasible to carry out surgical treatment at the same time.

Objective to investigate the relationship between serum resistin levels and type 2 diabetes mellitus (T2DM) complicated with peripheral neuropathy (PN). Methods the serum resistin levels in the following two groups were determined by enzyme-linked immunoassay (ELISA). 96 patients with T2DM were divided into two groups: DM complicated with PN group (DM + PN, n =52), DM alone group (DM, n =44) and control group (NC, n =34). Results the serum resistin levels in DM + PN and DM groups were significantly higher than those in NC group (P<0.05); Compared with the two groups, the serum resistin level in DM + PN group was significantly higher than that in DM group (P<0.05). Correlation analysis showed that the serum resistin concentration was significantly positively correlated with fasting serum insulin (FINS), fasting serum endothelin (ET), systolic blood pressure (SBP), diastolic blood pressure (DBP) and HOMA insulin resistance index (HOMA-IR) (all P<0.05), and significantly negatively correlated with HOMA-β cell function index (HOMA-β) and fasting serum nitric oxide (NO) (P<0.05). Conclusion The serum resistin level in DM + PN group was significantly higher than that in DM group. The correlation between hyperresistinemia and ET and NO suggests that resistin may play a certain role in the pathogenesis of type 2 diabetic peripheral neuropathy (PN).

Objective to investigate the efficacy and safety of upper thoracic epidural block (HTEA) in the treatment of ischemic cardiomyopathy heart failure (CHF). Methods 117 patients with ischemic cardiomyopathy heart failure (CHF), including 90 males and 27 females; Patients aged 45-81 years. Enrolled patients: stenosis of at least one coronary artery and its branches ≥50% confirmed by coronary angiography; Cardiac function NYHA grade Ⅱ ~ Ⅳ; The left ventricular ejection fraction (LVEF) was ≤45%. The patients were randomly divided into HTEA group and control group: 63 cases in HTEA group, 51 males and 12 females; In the control group, 54 patients, 39 males and 15 females, were treated with conventional drugs for heart failure. In the HTEA group, the total effective rate was 95.2%, which was higher than that in the control group (85.2%, P<0.05). After treatment, the mitral and tricuspid valve regurgitation area decreased significantly in the HTEA group, and the mitral valve regurgitation area decreased from (6.6 ± 3.7) cm2 to (4.3 ± 2.7) cm2, and the tricuspid valve regurgitation area decreased from (5.4 ± 3.2) cm2 to (3.3 ± 2.3) cm2 (P<0.01). The LVEF increased from (38 ± 12) % to (49 ± 10) %, which was significantly higher than that in the control group (P<0.01). The left ventricular end diastolic volume (LVEDV) and end systolic volume (LVESV) in the HTEA group LVESV decreased from (215±99) ml to (147±73) ml (P<0.01). The improvement of the above indexes in the control group was not obvious. HTEA did not improve the diastolic function indexes significantly (P>0.05). After treatment, the plasma BNP level of patients in the HTEA group was significantly lower than that in the control group ([678±521) ng/L vs 237±225) ng/L] (P<0.05). Conclusion the combination of HTEA in the treatment of ischemic cardiomyopathy heart failure is effective and safe, and can significantly improve the cardiac function.

Objective to compare the efficacy of minimally invasive thoracic catheter (central venous catheter) drainage and traditional multiple thoracic puncture and aspiration in the treatment of pneumothorax.Methods Forty-six patients with spontaneous pneumothorax with first onset were randomly divided into two groups from March 1, 2005 to August 31, 2006, and received central venous catheter drainage (minimally invasive catheter placement group) and multiple thoracic puncture and aspiration (multiple thoracic puncture group) respectively.The efficacy was evaluated by the treatment efficiency and the remaining pneumothorax volume after treatment, and the economy was evaluated by the cost of main consumables, the average length of hospital stay and the number of main treatment measures. Results The effective rate of treatment was 100% in the minimally invasive catheter placement group and 91.30% in the multiple thoracic puncture group; The average remaining pneumothorax volume at discharge was 4.17% in the minimally invasive catheterization group and 18.62% in the multiple thoracic puncture group; At 1 week follow-up, the average residual pneumothorax volume was 0.53% in the minimally invasive catheterization group and 7.59% in the multiple thoracic puncture group. In the minimally invasive catheterization group, the consumables such as central venous catheter, drainage bottle and reexamination chest X-ray film were used, and the per capita consumables cost was 264.79 yuan; Compared with the multiple thoracic puncture group, the average cost of consumables such as puncture kit and reexamination of chest radiograph was 233.62 yuan. Compared with the multiple thoracic puncture group, the average hospitalization days and infection prevention days were reduced by 2.36 and 2.13 days, respectively, and the average oxygen inhalation time was reduced by 36.7 h. Conclusion The efficacy of minimally invasive thoracic tubing was significantly better than that of multiple thoracic puncture and air pumping, and the total hospitalization cost was reduced.

Objective to investigate the changes of serum S100B protein and neuron-specific enolase (NSE) concentration in patients undergoing cardiac valve replacement under cardiopulmonary bypass (CPB) and the effects of cerebroside carnosine on the concentration of S100B protein and NSE. Methods 40 patients undergoing cardiac valve replacement under CPB were randomly divided into cerebroside carnosine group (Group A) and control group (Group B), 20 cases in each group. Group A was given intravenous injection of cerebroside carnosine 20 ml (50 μ g of ganglioside per ml and 3.2 mg of polypeptide per ml) after anesthesia induction, and group B was given equal volume of normal saline. Blood of internal jugular vein bulb was collected at each time point before (T1), at the end of CPB (T2) and 2 h (T3), 6 h (T4) and 24 h (T5) after CPB, respectively. Serum S100B protein and NSE concentrations were determined by immunosandwich double antibody enzyme-linked immunosorbent assay (ELISA). Results The concentrations of S100B protein and NSE in both groups reached the peak at the end of CPB, and then continued to decrease. The concentration of NSE in group B was also significantly higher than that in group A at T2, T3 and T4 time points (P<0.05). Conclusion Cerebroside carnosine can reduce the release of S100B protein and NSE in patients undergoing cardiopulmonary bypass heart valve replacement, and may have a certain brain protective effect.

Objective to investigate the clinical efficacy of percutaneous intervertebral disc electrotherapy (IDET) combined with intraspinal epidural steroid injection in the treatment of discogenic low back and leg pain. Methods the clinical efficacy of 59 patients with discogenic low back and leg pain was retrospectively analyzed by IDET combined with CT-guided intraspinal epidural steroid injection (Limedasone). The efficacy was evaluated by WHO pain relief criteria, and the data obtained were statistically compared with the clinical efficacy of 38 cases treated with IDET alone. Results The effective rate of IDET after follow-up was 71% within 1 week, 56% between 1 week and 1 month, 81% between 1 and 3 months, and 89% between 3 and 6 months. The difference of pain relief between IDET combined with intraspinal epidural steroid injection and IDET alone between 1 week and 1 month was statistically significant (χ 2=4.375, P<0.05). Conclusion The four clinical stages of the rehabilitation process after IDET are in line with the histological changes after intervertebral disc hyperthermia, The use of IDET combined with intraspinal epidural injection of steroids can relieve the pain of patients in the edema period after IDET and improve the curative effect.

Objective to investigate the expression of multidrug resistance associated protein (MRP), glutathione-S-transferase π (GST-π), DNA topoisomerase Ⅱ α (TOPOⅡ α) and cyclooxygenase-2 (COX-2) in ovarian cancer tissues and their relationship with drug resistance and prognosis. Methods Sixty patients with epithelial ovarian cancer admitted to Peking Union Medical College Hospital from 2000 to 2004 were selected and treated with platinum-based and paclitaxel first-line chemotherapy for 6 to 9 courses after cytoreductive surgery. The follow-up data were complete. Among them, 35 cases were serous breast carcinoma, 2 cases were mucinous cystadenocarcinoma, 11 cases were clear cell carcinoma, 4 cases were transitional cell carcinoma and 8 cases were endometrioid carcinoma. The average age of the patients was 51.2 years old. The expressions of MRP, GST-π, Topo Ⅱα and COX-2 in ovarian cancer samples were detected by immunohistochemistry, and the relationship between their expressions and drug resistance and prognosis of ovarian cancer patients was analyzed. Results The positive expression rates of MRP, GST-π, Topo Ⅱα and COX-2 in ovarian cancer samples were 26.7%, 93.3%, 61.7% and 78.3%, respectively. The positive rate of Topo Ⅱα in stage Ⅲ ~ Ⅳ was significantly lower than that in stage Ⅰ ~ Ⅱ (56.6% vs 78.6%, P<0.05). MRP positive, GST-π strong positive, Topo Ⅱα negative and COX-2 strong positive expression were considered as four risk factors related to recurrence, drug resistance and poor prognosis. The number of patients with ≥2 risk factors in the recurrence group was significantly more than that in the non-recurrence group (81.6% vs 54.5%, P<0.05). The number of patients with ≥2 risk factors in the chemoresistant group was significantly more than that in the chemosensitive group (100% vs 55.6%, P<0.01). The 3-year survival rates of patients with ≥2 risk factors and those with less than 2 risk factors were 69.9% and 91.7%, respectively (P>0.05). Conclusion MRP, GST-π, Topo Ⅱ α and COX-2 expressed in moderate to high degree in ovarian cancer tissues. More than two risk factors of MRP positive, GST-π strong positive, Topo Ⅱα negative and COX-2 strong positive can be used as important indicators to predict recurrence and drug resistance in ovarian cancer patients.

Objective to investigate the relationship between thiopurine methyltransferase (TPMT) genotype and azathioprine tolerance in patients with rheumatism. Methods Four common TPMT mutation alleles, TPMT*2 (G238C), TPMT*3A (A719G/G460A), TPMT*3B (G460A) and TPMT*3C (A719G), were detected by allele specific polymerase chain reaction (AS-PCR) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 200 patients with rheumatism. 194 patients were treated with azathioprine (dose 50~150 mg/d) and followed up for 3 months. Results 4 cases of TPMT*3C heterozygotes were detected in 200 patients with rheumatism, but no TPMT*2, TPMT*3B and TPMT*3A mutations were detected. The genotype frequencies were 98% for homozygotes of wild type gene and 2% for heterozygotes of TPMT*1/TPMT*3C. The frequency of mutation alleles in rheumatism patients was 1%. The mean value of TPMT activity of 4 cases of TPMT*3C heterozygotes was (2.4±1.2) U/ml erythrocytes, and the mean value of TPMT activity of 196 cases of homozygotes of wild type gene was (12.2±6.8) U/ml erythrocytes. The mean value of enzyme activity of TPMT*3C heterozygotes was significantly lower than that of homozygotes of wild type gene, and the difference was statistically significant. Among 194 patients treated with azathioprine, 18 cases had bone marrow suppression, 2 cases had severe hematopoietic crisis, 6 cases had liver function impairment, 4 cases had TPMT*3C heterozygotes Severe hematopoietic crisis may occur. Detection of TPMT genotype before medication is important to improve the safety of treatment.

Objective to investigate the effects of 17 β-estradiol (E2) on the activities of inducible nitric oxide synthase (iNOS) and endothelial nitric oxide synthase (eNOS) in myocardial ischemia-reperfusion, and to explore the mechanism of myocardial protection.Methods Forty ovariectomized (OVX) rats were randomly divided into control group (group C) using Langendorff isolated rat heart perfusion model: the isolated rat heart was pre-perfused for 15 min; Ischemia-reperfusion group (I-R group): Modified St. Thomas Ⅱ cardioplegia was perfused to complete the whole process of myocardial ischemia-reperfusion; Solvent control group (group D): 0.1% dimethyl sulfoxide (DMSO) in cardioplegia solution, the same as I-R group; The activity of iNOS and eNOS in myocardium was measured before and after ischemia-reperfusion, and the contents of creatine phosphokinase (CPK), lactate dehydrogenase (LDH), nitric oxide (NO) in coronary effluent and the changes of cardiac function were measured. The effect of E2 on myocardial ischemia-reperfusion injury (MIRI) was observed. Results After myocardial ischemia-reperfusion, the activity of iNOS decreased (P<0.01), and the activity of iNOS increased (Group C (8.9±3.7) nmol·min-1·g-1, Group I-R (15.8±2.4) nmol·min-1·g-1, Group D (17.6±3.2) nmol·min-1·g-1, P<0.01), and the activity of iNOS increased more significantly in Group E [(25.85±5.21) nmol·min-1·g-1, P<0.01]. The production of NO decreased in Group I-R and Group D (both P<0.05), and NO increased in Group E [Group C (31±5) μ mol/L, Group E (33±6) μ mol/L, P

Objective to observe whether transplanted neural stem cells can successfully pass through the blood-brain barrier of transgenic mice with Alzheimer's disease (AD), and to observe the survival and migration in their brains. Methods The neural stem cells were isolated and cultured in vitro, labeled with green fluorescent protein and transplanted through the tail vein of AD mice. Serial frozen sections of brain were taken at 48 h, 1 week, 2 weeks and 1 month after transplantation, and the distribution and migration of transplanted cells were observed under fluorescence microscope. Results At different time points after transplantation, green fluorescent protein positive cells were scattered in the brain, and the distribution was more common around blood vessels within 48 h, and it was extended to 4 weeks with time, and the diffuse distribution was also observed in the hippocampus and cerebral cortex. However, the transplanted mice grew well within 4 weeks, and no tumor formation was observed. Conclusion Neural stem cells can successfully pass through the blood-brain barrier of transgenic mice with Alzheimer's disease, diffuse in their brains and migrate with time.

Objective to observe the expression of hypoxia-inducible factor (HIF) 1 α in esophageal squamous cell carcinoma and its relationship with chemotherapy and clinical prognosis. Methods 48 patients with advanced esophageal squamous cell carcinoma were treated with cisplatin/paclitaxel-based combination chemotherapy, paclitaxel 175mg/m2, intravenous drip for 3 h on the first day; Cisplatin 80 mg/m2, on the 2nd to 4th day, 21 days as one cycle, each case completed at least 2 cycles of chemotherapy, and the efficacy evaluation was clear.The above case specimens were from the Department of Pathology, the First Affiliated Hospital of Zhengzhou University.The expression of HIF-1a in esophageal cancer tissues was detected by immunohistochemistry.Another 20 cases of normal esophageal mucosa were taken as controls.X2 test and Kaplan-Merier method were used for statistical analysis.Results The positive staining of HIF-1a in esophageal squamous cell carcinoma tissues was mainly located in the cytoplasm/nucleus of cancer cells, and the positive rate was 43.75% (21/48), while the normal esophageal mucosa was negative. The chemotherapy effective rate of HIF-1a positive expression was 23.81% (5/21), which was significantly lower than that of HIF-1a negative expression 70.37% (19/27) (P<0.05). The median disease progression time, median survival time and 2-year survival rate of HIF-1a positive expression were 2.0 months, 6.0 months and 13.3%; The median disease progression time, median survival time and 2-year survival rate of HIF-1a negative expression patients were 5.0 months, 11.0 months, and 42.2%. The difference between the two groups was statistically significant (P<0.05). Conclusion Immunohistochemical detection of HIF-1a protein expression can be used as one of the clinical indicators to predict the efficacy of chemotherapy and clinical prognosis of esophageal cancer.

Objective to construct pcDNA3.1-vascular endothelial growth factor (VEGF) 165 plasmid and observe its effect on bone defect repair in rabbits. Methods the recombinant eukaryotic expression plasmid pcDNA3.1-VEGF165 was constructed. Thirty New Zealand white rabbits were used to prepare bilateral radial bone defect model. The control group was treated with the same amount of normal saline instead of plasmid solution and treated in the same manner. X-ray photographs were taken at 1, 2, 4, 6, 8 and 12 weeks after operation, and the local tissue repair and neovascularization were observed, and the microvessel density (MVD) was calculated. Results the pcDNA3.1-VEGF165 plasmid was successfully constructed and sequenced correctly. HE staining: in the experimental group, a large number of neovascularization and blood flow were formed at 2 weeks, a large number of osteoblasts proliferated and trabecular bone formed at 4 weeks, and the cortical bone reconstruction was completed at 12 weeks, and the bone marrow cavity was recanalized; The MVD of the control group and the experimental group were 56.1±6.1 and 69.1±5.4 at 2 weeks, respectively, compared with 42.2±6.4 and 47.0±7.5 at 1 week, and the differences between groups and within groups were statistically significant (t =8.0347, P =0.0000). Conclusion the direct application of PcDNA3.1-VEGF165 plasmid solution can express and upregulate VEGF165, which can promote the formation of local microvessels and accelerate the repair of bone defects.

Objective to study the effect of RNA interference technique on the expression of aquaporin 1 (AQP1) gene in rat pleural mesothelial cells, and to explore the feasibility of using this technique in the treatment of pleural effusion.Methods Rat pleural mesothelial cells were cultured in vitro.After cell identification, the expression of AQP1 was detected by immunocytochemistry and reverse transcription-polymerase chain reaction (RT-PCR). Two recombinant short hairpin RNA (shRNA) plasmids targeting rat AQP1 gene were designed and constructed. After liposome transfection of rat pleural mesothelial cells for 48 h, the inhibitory effects of AQP1 gene expression were detected by RT-PCR and Western blot at mRNA and protein levels, respectively. Results AQP1 expression existed in pleural mesothelial cells, and the two recombinant AQP1-siRNA plasmids designed could inhibit the expression of AQP1 in pleural mesothelial cells to varying degrees, and the inhibition rates at mRNA level were 83.5% and 90.9%, respectively; The inhibition rate at protein level was 41.2% and 67.6%, respectively, compared with the blank control group and the negative control group, the difference was statistically significant (P<0.01). Conclusion Plasmid-mediated RNA interference can significantly inhibit the expression of AQP1 gene in rat pleural mesothelial cells, and the inhibition rate is sequence-related, which is a potential new method for gene therapy of pleural effusion.

Objective to investigate the role of extracellular signal-regulated kinase (ERK) and transforming growth factor β 1 (TGF-β 1) in asthma airway remodeling, and to investigate the regulation of glucocorticoid on ERK, TGF-β 1 and asthma airway remodeling. Methods Thirty SD rats were randomly divided into control group, asthma group and dexamethasone intervention group (DM group). The phosphorylated ERK (P-ERK) in lung tissue was determined by immunohistochemistry, and the content of TGF-β 1 in serum was determined by ELISA. Rat airway epithelial cells were cultured in vitro with BB-type platelet-derived growth factor (PDGF-BB), U0126 and budesonide (BUD) as tool drugs. The phosphorylation level of ERK in cells was detected by Western blot, and the content of TGF-β 1 in cell supernatant was detected by ELISA. Results The mean absorbance of P-ERK and the content of TGF-β 1 in asthma group were (31.1±2.2) and (28.1±7.4) μ g/L, respectively Both U0126 and BUD inhibited phosphorylation of ERK; Conclusion ERK phosphorylation and TGF-β 1 play an important role in airway remodeling of bronchial asthma. PDGF-BB could not induce the release of TGF-β 1 from normal rat tracheal epithelial cells through ERK phosphorylation; Glucocorticoid could inhibit ERK phosphorylation.

Avian influenza virus has always been believed to be infected and prevalent only in poultry. Because of its species barrier and receptor specificity, it is rare to infect humans and cause morbidity and death [1]. In recent years, many cases of avian influenza virus subtypes H5N1 [2-3], H7N7 [4], H7N3 [5] and H9N2 [6] have been reported worldwide to infect humans and cause morbidity and death.

Asthma remains a chronic disease that threatens public health because asthmatic airway inflammation persists, and the airway responds increasingly to various stimuli and causes asthma attacks. Medication can reduce airway inflammation and alleviate or prevent the symptoms of airway stenosis.

A 16-year-old male was admitted to the hospital on April 21, 2006 due to intermittent chest pain for 6 months. Physical examination showed brachial artery blood pressure 144/63 mm Hg (1 mm Hg =0.133 kPa), no cyanosis on the lips, continuous machine-like murmur in the second intercostal area at the left margin of the sternum, enhanced and split second heart sound in the pulmonary artery area, and positive shooting sound in the femoral artery.

The patient was a 37-year-old young woman with previous health. He was admitted to hospital due to "weakness of limbs with numbness for 1 week". No abnormalities were found in medical examination. The physical examination of nervous system showed clear consciousness, good spirit, clear speech, soft neck, negative Klinefelter sign, no exception in cranial nerve examination, muscle strength of limbs V-grade, glove-sock-like shallow sensation of limbs, deep sensation existence and symmetry, tendon reflex of limbs disappeared, and pathological signs were negative.