Volume 85  ·   Issue 38  ·  2005

Hepatic encephalopathy is a syndrome of metabolic disorders and dysfunction of central nervous system caused by severe liver disease. From the record up to now, after more than 100 years of research, there are still many unclear problems and even misunderstandings.

Objective to investigate the preparation method of porcine bladder acellular matrix and its feasibility as biological scaffold material in tissue engineering bladder construction. Methods Fresh porcine bladder was acellularized by enzyme digestion and detergent washing, HE staining and electron microscopy examination respectively. Cytotoxicity was determined by MTT assay. After most of New Zealand rabbits underwent cystectomy, xenogeneic bladder acellular matrix was used to repair, and the regeneration of repaired tissue was observed after 6 weeks. Results Both methods could remove cellular components in the bladder, and the cytotoxicity was grade 1 and the cellular compatibility was good after light microscopy and electron microscopy.

Objective to investigate the reversal effect of recombinant adenovirus-mediated multidrug resistance associated protein (MRP) antisense RNA on the multidrug resistance phenotype of human hepatocarcinoma cell line SMMC-7721/ADM induced by doxorubicin in vitro.Methods A self-constructed recombinant adenovirus (Ad-Asmrp) carrying antisense MRP was used to transfect human hepatocarcinoma cell lines in vitro.The median lethal dose (IC50) of transfected cells to doxorubicin (ADM) and daunorubicin (DNR) was measured and the resistance factor (RF value) was calculated. The changes of DNA uptake and P190 expression on cell membrane surface were dynamically measured by flow cytometry at different time points after transfection, and the changes of MRP mRNA levels were reflected by RT-PCR. Results In vitro experiments, the IC50 of ADM and DNR in human hepatoma resistant cells transfected with recombinant adenovirus carrying MRP antisense RNA was 0.487 μ g/ml and 0.328 μ g/ml, respectively, and the RF values were 97.4 and 109.3, respectively, and the RF values decreased by 36.8 and 35.4, respectively. At 24 hours after transfection, the MRP mRNA level began to decrease and showed a continuous decreasing trend (P<0.01), and the expression of P190 protein continued to decrease after 48 hours (P<0.01), and the two trends were consistent. The intracellular DNR concentration was significantly increased by flow cytometry (P<0.01) at 48 hours after transfection. Conclusion MRP antisense RNA mediated by recombinant adenovirus can effectively block the expression of MRP gene, increase the sensitivity of human hepatoma resistant cell lines to chemotherapy drugs in vitro experiments, and increase the sensitivity of human hepatoma resistant cell lines to chemotherapy drugs

Objective to investigate the abnormal development of cardiac cone in Cx43 gene-deficient mice. Methods C57/BL6 mice were selected from embryonic day (E) 11.5 d to 1 d after birth as the research objects. According to the genotypes, they were divided into homozygous (Cx43- / -), heterozygous (Cx43+ / -) and wild type (Cx43+ / +). The genotypes were identified by polymerase chain reaction method. The expressions of rhabdomuscle actin α-SCA, smooth muscle actin α-SMA and AP-2 α in neural crest cells were determined by immunohistochemistry; The expression of AP-2 α mRNA was detected by in situ hybridization. Results Cx43- / -died within 24 h after birth. The abnormal bulge of the cone of the right ventricular outflow tract was revealed by gross anatomy. HE staining showed a large number of abnormal trabecular tissue proliferation processes in the right ventricular outflow tract wall, and the right ventricular outflow tract cavity was obviously narrowed. There was no obvious abnormality of Cx43+ / -. The expression of α-SCA in the central region of the proximal outflow tract of Cx43- / -was obviously lagged. Compared with Cx43+ / +, the expression of α-SMA in the right ventricular outflow tract of Cx43+ / -and Cx43- / -mice was stronger than that of Cx43+ / +, mainly located in the abnormal proliferation site of the right cone stem. The expression of AP-2 α protein and its mRNA in the E13.5 outflow tract septum of Cx43- / -mice was increased, and the expression position was abnormal. Conclusion Cx43 KO mice were characterized by obstructive malformation caused by abnormal proliferation of the right ventricular outflow tract.

Objective to investigate the analgesic effect of intrathecal transplantation of immortalized rat astrocytes (IAST/hPPE) on chronic neuralgia in rats.Methods 40 adult male SD rats were randomly divided into four groups: Naive group, SNI group, SNI + IAST group and SNI + IAST/hPPE group; Except for the Naive group, the right lower extremity preserved nerve injury model (SNI) was established; One week later, immortalized rat astrocyte (IAST) or IAST/hPPE cell lines co-cultured with 5' bromo-2-deoxyuridine (BrdU) were intrathecally transplanted into the spinal cord at L4-6 level. The difference of 50% foot contraction threshold (PWT) on the left and right sides of rats in each group and the effect of intraperitoneal injection of naloxone on their analgesic effect were measured every week. The levels of leucine enkephalin (L-EK) in spinal cord and the expression of BrdU in transplanted cells were detected by immunohistochemistry and radioimmunoassay. Results the abnormal pain in the right foot of rats appeared 1 week after SNI. Compared with the Naive group, the difference of 50% PWT on the left and right sides of the other three groups was significantly increased (P<0.01); In SNI + IAST/hPPE group, the symptoms of dysalgesia were significantly reduced, while in SNI group and SNI + IAST group, there was no significant change. The difference of left and right 50% PWT in SNI + IAST/hPPE group was significantly lower than that in SNI group and SNI + IAST group (P<0.01); The analgesic effect of SNI + IAST/hPPE group was antagonized by naloxone; Compared with the Naive group, the content of L-EK per mg of spinal cord in the other three groups was significantly higher (P<0.01), SNI + IAST/hPPE group (108.1 pg/mg ± 12.5 pg/mg) was significantly higher than SNI group and SNI + IAST group (P<0.01), while the difference between SNI group and SNI + IAST group was not statistically significant (25.4 pg/mg ± 1.9 pg/mg vs 28.0 pg/mg ± 2.1 pg/mg, P>0.05); Conclusion Intrathecal transplantation of IAST/hPPE cells can inhibit the allodynalgesia in rats with chronic neuralgia, which is related to the effect of enkephalin secreted by transplanted cells on opioid receptors.

Objective to investigate the cloning, tissue distribution and expression of novel human glioma-related LNX genes in different grades of astrocytoma, and the molecular mechanism of their involvement in glioma signaling pathway. Methods Fetal brain cDNA library was constructed and sequenced to obtain the novel full-length LNX gene; Human multi-tissue library (MTC) was used as a template to study the distribution of LNX in normal human tissues; The expression of LNX in cerebral astrocytoma was detected by microarray containing 13 939 human gene expression profiles; Northern hybridization to verify the expression of LNX in different grades of glioma; Results the novel LNX gene cloned from human fetal brain library was about 3.7kb long, contained an open reading frame of 1899bp, encoded a 632 amino acid protein with a molecular weight of about 70kDa, accession number AF237782; LNX was highly expressed in brain, kidney and pancreas, as well as in heart, placenta, lung, liver, spleen, thymus and prostate; However, the expression of LNX in glioma tissues was decreased, which was closely related to tumorigenesis; The interaction between LNX and Numb and SKIP protein in the Notch signaling pathway was detected by yeast two-hybrid screening. The interaction between LNX and SKIP protein can affect the subcellular localization of Numb and the binding site of Numb in the Notch signaling pathway. Conclusion The expression profiling microarray can quickly and efficiently lock the genes closely related to glioma and find tumor markers. LNX is closely related to glioma, and it can participate in the development of glioma through Notch signaling pathway, which can be a potential therapeutic target for glioma.

Objective to select appropriate inhalation anesthetic drugs and concentrations for short incubation period somatosensory evoked potential (SSEP) monitoring under the monitoring of bispectral index of brain electricity (BIS). Methods Sixty patients undergoing elective neurosurgery were randomly divided into enflurane, isoflurane and desflurane groups (20 cases in each group). Different concentrations of anesthetics were inhaled according to the minimum alveolar effective concentration (MAC) of 0.3, 0.5, 0.75, 1.0 and 1.5 respectively. The N20 amplitude and incubation period of SSEP were recorded, and BIS monitoring was performed at the same time. The effects of three inhaled anesthetics on SSEP and BIS were observed at different inhalation concentrations. Results With the increase of inhalation concentration, the N20 amplitude of SSEP decreased gradually and the incubation period of SSEP gradually prolonged in all three groups. At 1.5 MAC, the N20 waveforms of 3 subjects in the isoflurane and desflurane groups disappeared, and the waveforms of 6 subjects in the enflurane group disappeared. BIS monitoring found that at 1.0 MAC, the BIS values of each group were below 60, while at 0.75 MAC, the BIS values of some patients in the enflurane group (BIS values: 45~64) and desflurane group (44~61) were still higher than 60, while the BIS values in the isoflurane group were below 60 (39~58). There was no good correlation between the amplitude and incubation period of cortical SSEP and BIS values. Conclusion The effects of volatile inhalation anesthetics on BIS and cortical SSEP are dose-related. Under the appropriate anesthesia depth, 0.75 MAC of isoflurane is more suitable for intraoperative cortical SSEP monitoring. Although the effects of volatile inhalation anesthetics on EEG and SSEP are related to their effects on cerebral cortex, respectively, the correlation between them is poor.

Objective to investigate the relationship between the change trend of flash visual evoked potential (fVEP) N2 wave and the change of intracranial pressure before and after mannitol application. Methods Fifty-two patients with hypertension were selected and divided into mannitol alone group and sodium glycerol chloride combined group, 32 cases and 20 cases, respectively. The N2 wave incubation period and amplitude of fVEP were measured by NIP-200 noninvasive intracranial pressure monitor before and after the 1st, 4th and 10th mannitol application, and compared. Results The N2 wave incubation period in the mannitol alone group began to shorten at 30 minutes after mannitol application, and shortened to the shortest at 2 hours after application. The incubation period of N2 wave rose at 4 hours, but was still shorter than that before mannitol application. The amplitude of N2 wave did not change significantly. With the increase of mannitol application times, the shortening degree of N2 wave incubation period decreased gradually. The incubation period of N2 wave in the group with sodium glycerol chloride combined with mannitol application did not change significantly before and after mannitol application. Conclusion The incubation period of N2 wave of fVEP changed before and after mannitol application, suggesting that the change of incubation period of N2 wave of fVEP can reflect the change trend of intracranial pressure.

Objective to investigate the clinical significance of the changes of plasma adrenomedullin (ADM) and UⅡ (UⅡ) in patients with congenital heart disease (CHD) complicated with pulmonary hypertension (PH) before and after operation. Methods 52 patients were divided into 3 groups according to pulmonary artery systolic blood pressure: no PH group (<30 mm Hg), 18 cases in mild PH group (30~49 mm Hg) and 17 cases in moderate to severe PH group (≥50 mm Hg). The levels of ADM and UⅡ in the three groups were measured before, immediately after and 7 days after operation, and the changes before and after operation were compared. Results (1) There was a positive correlation between pulmonary artery pressure (PAP) and plasma ADM concentration (r =0.8012, P<0.01; Immediate postoperative r =0.6325, P<0.01; 7d after operation r =0.7126, P<(2) There was no correlation between the concentration of UⅡ and PAP in the three groups (all P>0.05). (3) The preoperative ADM concentration in the group without PH was 33 pg/ml ± 5 pg/ml, 29 pg/ml ± 4 pg/ml immediately after operation, and 20 pg/ml ± 3 pg/ml 7 days after operation; The concentration of ADM in the mild PH group was 44 pg/ml ± 8 pg/ml before operation, 40 pg/ml ± 6 pg/ml immediately after operation, and 34 pg/ml ± 4 pg/ml 7 days after operation; The concentrations of ADM were 60 pg/ml ± 10 pg/ml before operation, 58 pg/ml ± 8 pg/ml immediately after operation, and 38 pg/ml ± 4 pg/ml on 7 days after operation in the moderate to severe PH group. The ADM concentrations in each group showed a decreasing trend after operation, but the difference was statistically significant only on 7 days after operation and before operation (no PH group q =5.41, P<0.01; q =4.76 in mild PH group, P<0.01; q =6.32 in moderate to severe PH group, P<(4) The concentration of UⅡ in the group without PH was 2.2 pmol/L±0.5 pmol/L before surgery, 2.2 pmol/L±0.44 pmol/L immediately after surgery, and 2.2 pmol/L±0.6 pmol/L 7 days after surgery; In the mild PH group, the concentration of UⅡ was 2.7 pmol/L±0.6 pmol/L before operation, 2.6 pmol/L±0.6 pmol/L immediately after operation, and 2.6 pmol/L±0.5 pmol/L 7 days after operation; The concentration of UⅡ in the moderate to severe PH group was 2.9 pmol/L±0.6 pmol/L before operation, 2.6 pmol/L±0.7 pmol/L immediately after operation, and 2.8 pmol/L±0.4 pmol/L 7 days after operation. There was no statistically significant difference in the concentration of UⅡ in the three groups before and after operation (all P>0.05). Conclusion (1) ADM plays an important role in PH formation and vascular remodeling. (2) There is no correlation between UⅡ and PAP, but it cannot be excluded that UⅡ plays an important role in PH formation and vascular remodeling. (3) Plasma ADM level can be used as one of the indexes to judge the severity of PH.

Objective to report the clinical and genetic characteristics of Waardenburg syndrome type Ⅱ in Chinese. Methods the family was followed up by the information provided by probands, and the detailed medical history investigation and general and specialist examination of all patients and some normal persons in the family were carried out by filling in questionnaire questionnaire, and the genetic map of the family was drawn. Results the clinical manifestations of each family varied greatly, and the clinical symptoms mainly included sensorineural deafness, iris heterochrome, facial freckles and early gray hair. The deafness could be unilateral or bilateral, prelingual deafness or postlingual deafness. The genetic map of the family showed that it was autosomal dominant inheritance with incomplete manifestation, and the deafness had a certain genetic tendency to appear early. According to the complete clinical data, the family was diagnosed as Waardenburg syndrome type Ⅱ. Conclusion The phenotypic characteristics of deafness and pigment abnormalities in this Chinese family have certain particularities. The complete clinical data of this family provided a basis for the formulation and improvement of the diagnostic criteria of Waardenburg syndrome type II in Chinese. It is also a supplement to the international clinical diagnostic criteria for WS.

Objective to investigate the application of cross-breakpoint fluorescence polymerase chain reaction (PCR) technique in the preimplantation genetic diagnosis (PGD) of α thalassemia (α thalassemia). Methods Single lymphocytes from Southeast Asian carriers of α thalassemia were obtained, and a stable cross-breakpoint fluorescence PCR technique was established for detecting α thalassemia. Results the average amplification efficiency of single lymphocytes was 90.0% (72/80), and the average allele disconnection (ADO) rate was 8.3% (6/72). After 4 cycles of PGO, 38 embryos were detected in 4 couples, and 38 blastomeres were obtained, of which 34 blastomeres were successfully amplified. The amplification efficiency was 89.5% (34/38), and the ADO rate was 5.9% (2/34). By PCR analysis, 11 normal embryos, 8 heterozygous embryos and 15 thalassemia major embryos were obtained. Eleven embryos were transplanted, and 2 clinical pregnancies were obtained. At 17 weeks of gestation, it was confirmed that they were completely normal embryos and heterozygous embryos by umbilical cord blood puncture, respectively. Two healthy male infants were born. Conclusion the application of single cell fluorescence PCR technique can be used to diagnose alpha thalassemia before implantation and achieve the goal of eugenics.

A 53-year-old male was admitted to the hospital on 24 November 2004 due to "multiple joint swelling and pain for 8 months, cough and shortness of breath for 2 months". In April 2004, the patient suffered from pain and swelling in the proximal interphalangeal joints of both hands and the first toe joints of both feet without inducement, accompanied by morning stiffness for half a day.The erythrocyte sedimentation rate (ESR) was 26 mm/h, rheumatoid factor (RF) was positive, and the diagnosis was rheumatoid arthritis. 　　

diabetic peripheral neuropathy (DPN) is the most common chronic complication of diabetes mellitus, and its incidence rate is as high as 60% ~90% [1]. DPN is mainly manifested as numbness, pain and paresthesia in the glove-like distribution area of the extremities, which seriously affects the quality of life of patients. The pathogenesis of this disease is not completely understood and there is no method to prevent it. It is also the main cause of foot ulcer, infection and amputation [2]. At present, besides strict control of blood sugar, there is still a lack of specific clinical treatment methods with satisfactory curative effect. Since 2004, peripheral nerve release has been used in our hospital to treat 57 patients with DPN, and the effect is obvious. The report is as follows.

World's authoritative magazine<Nature>and the prestigious American Institute of Scientific Information (ISI) published, influential in the fields of scientometrics and bibliographic analysis<Scientific Observation, published in July this year<Asia challenges European leadership in science>and<The two articles show that the Eastern Hemisphere accounts for an increasing share of scientific achievements, and Asian countries are catching up with Europe and the United States.

A 25-year-old female was admitted to the hospital on February 22, 2005 due to "51 d of menopause, vaginal bleeding with lower abdominal distension for 6 d". The outpatient was admitted to the hospital with "ectopic pregnancy". Usually, her menstruation was irregular, often delayed by 1 week. The last menstruation was on January 2, 2005, with moderate amount, clean for 5 d and normal color. Before 6 d, she had irregular vaginal bleeding, small amount, dark red color, and slight paroxysmal distension and pain in the lower abdomen. The urine examination of admission was positive for human chorionic gonadotropin (hCG), blood β-hCG 241 U/L, and serum progesterone 10 ng/ml. Transabdominal B-ultrasound showed that there was no gestational sac in the uterine cavity, and a mixed echo mass was seen in the left appendage, 3.9 cm ×2.4 cm in size, and "double ring sign" seemed to be visible inside.

A 56-year-old male was admitted to the intensive medical ward (ICU) of Peking Union Medical College Hospital on March 14, 2003 due to headache, diplopia for 5 d, left blepharoptosis with vomiting for 3 d, and disturbance of consciousness for 7 h. The patient had headache and diplopia without trigger on March 9th, and had difficulty lifting the left eyelid on March 11th, accompanied by nausea and vomiting. On March 13th, the patient came to our emergency department because of the aggravation of headache. CT and MRI of the head showed subarachnoid hemorrhage (SAH). The next day, he developed disturbance of consciousness and respiratory failure. Check FⅧ:C: 16.2%.