2007年 · 第120卷第13期

The surgical procedure of C7 root transfer from the healthy side was first performed successfully on August 26, 1986.1,2 Since then, this procedure has proved one of the major treatments for brachial plexus root avulsions.Nowadays, attitudes of doctors towards this operation have changed from doubt and surprise to deep conviction and acceptance.3,4 The latest progresses in experimental and clinical studies on contralateral C7 root transfer are reviewed in this article。

Abstract：Background Macrophage stimulating protein (MSP) is produced by human bone marrow endothelial cells. In this study,we sought to observe its effects on inducing the expansion of early hematopoietic progenitor cells which were cultured in a liquid culture system in the presence of the combination of stem cell factor (SCF), interleukin 3 (IL-3), interleukin 6 (IL-6), granulocyte macrophage-colony stimulating factor (GM-CSF), erythropoietin (EPO) (Cys) and MSP or of Cys and bone marrow endothelial cell conditioned medium (EC-CM).Methods Human bone marrow CD34+ cells were separated and cultured in a liquid culture system for 6 days.Granulocyte-macrophage colony forming unit (CFU-GM) and colony forming unit-granulocyte, erythrocyte, macrophage,megakaryocyte (CFU-GEMM) were employed to assay the effects of different treatment on the proliferation of hematopoeitic stem/progenitor cells. The nitroblue tetrazolium (NBT) reductive test and hoechest 33258 staining were employed to reflect the differentiation and apoptosis of the cells respectively.Results MSP inhibited the proliferation of CFU-GM and CFU-GEMM in semi-solid culture and the inhibitory effect on CFU-GEMM was stronger than on CFU-GM. MSP inhibited the differentiation of early hematopoietic progenitor cells induced by hematopoietic stimulators. Bone marrow (BM) CFU-GEMM was 2.3-fold or 1.7-fold increase or significantly decreased in either Cys+EC-CM, Cys+MSP or Cys compared with 0 hour control in liquid culture system after 6 days.Conclusion MSP, a hematopoietic inhibitor, inhibits the differentiation of early hematopoietic progenitor cells induced by hematopoietic stimulators and makes the early hematopoietic progenitor cells expand in a liquid culture system。

Abstract：Background Gene therapy has been a hot spot in repair of bone defects in recent years. This study aimed to construct a recombinant plasmid pcDNA3.1-VEGF165, and to observe the effect of vascular endothelial growth factor 165 (VEGF165)gene therapy on bone defects in rabbits.Methods Total RNA was extracted from rabbit bone tissues. VEGF165 cDNA fragment was prepared by reverse transcription and the gene was cloned by polymerase chain reaction (PCR). Plasmid pMD18-T/VEGF165 combined with pcDNA3.1 was cloned to reconstruct pcDNA3.1-VEGF165 plasmid. Thirty New Zealand white rabbits weighing (2.50±0.13)kg were used to establish models of bone defects (1 cm in length) of the bilateral radii. The bone defects were repaired with absorbable gelatin sponge. After the operation, physiological sodium chloride solution was injected into the injured site in one of the forelegs of the rabbits as the control group, and pcDNA3.1-VEGF165 plasmid (0.2 ml, 200 ng)was injected into the opposite foreleg as the experiment groups. At weeks 1, 2, 4, 6, 8, and 12 after the treatments, the bones were examined by X-ray, and the specimens of the bone defects were collected, stained with HE, and observed under a light microscope. The expression of VEGF165 mRNA was examined by real-time quantitative polymerase chain reaction (RQ-PCR).Results The pcDNA3.1-VEGF165 plasmid with a correct sequence was constructed successfully. Postoperative X-ray found no difference between the two groups at week 1. In the experiment group, callus and synostosis were observed after 2 weeks, and osteosis structure was normal at week 12; these phenomena occurred much later in the control group.In the experiment group, HE staining showed a large amount of newly formed blood vessels after 2 weeks, a number of bone trabeculae with osteoblasts proliferation at 4 weeks, and fresh bone cortex and reformed medullary cavity at 12 weeks; whereas in the control group these structures formed in later phases. The VEGF165 mRNA in the experiment group was expressed at a low level at week 1, reached the peak at weeks 3, and then decreased to a normal level after 6 weeks.Conclusions Local use of pcDNA3.1-VEGF165 plasmid at bone defects can upregulate the expression of VEGF165 and accelerate the formation of capillaries and the repair of bone defects. Angiogenesis and osteogenesis can be promoted by a combination of pcDNA3.1-VEGF165 and gelatin sponge。

Abstract：Background Peutz-Jeghers syndrome (PJS) is an autosomal dominantly inherited disease. STK11/LKB1 gene germline mutations have been identified as responsible for PJS. In our study, we investigated the molecular basis of PJS and evaluated correlation between the STK11 mutations and the Chinese population.Methods We collected three pedigrees of PJS and screened the 9 exons and their flanking intronic sequences of STK11/LKB1 gene in the probands and normal individuals in the families using polymerase chain reaction (PCR) and direct sequencing.Results Sequencing of the STK11 gene in the probands of 3 families revealed two novel mutations (c180C→G and c998-1002delGCAGC) in exon 1 and exon 8, respectively. The mutation of c180C→G resulted in a premature termination codon. The other mutation, a deletion of five nucleotides (998-1002delGCAGC) in exon 8, predicted to generate a translational frameshift and a termination at codon 1070.Conclusions The growing number of mutations in PJS pedigrees suggests the molecular basis of PJS. STK11 gene mutation can be detected in most patients with PJS。

Abstract：Background Transplanting a vascularized autologous submandibular gland (SMG) is considered an effective method to treat severe keratoconjunctivitis sicca. But the operation may fail due to the anatomic variances in the blood vessels of SMG. The present study aimed to investigate the submandibular glands at the microanatomy level.Methods The microanatomy of blood vessels including arteries and veins of submandibular gland was investigated using 30 adult corpses and 60 submandibular glands were anatomized under a surgical microscope. The lengths and diameters of the arterial and venous glandular branches were measured using sliding caliper.Results The submandibular gland was mainly supplied by the facial artery and submental artery, partly by the lingual artery and external jugular artery. The venous drainage of the submandibualr gland occurred through the anterior facial vein, the venae comitantes of facial artery, the vein close to the Whaston's duct (the hilum vein), and seldom drained to external jugular vein and other veins.Conclusions The anatomy of SMG is a complicated structure. Determining the main blood vessels of the submandibular gland is very important to achieve a successful vascularized autologous SMG transplant。

Abstract：Background It has been confirmed that the concentration of cefepime in cerebrospinal fluid (CSF) could reach the 10% of its concentration in plasma, exceeding the inhibitory concentration to 90% of organisms (MIC90) for common bacteria.However, the blood-brain barrier (BBB) penetration ability of cefepime is still unclear. The aim of this study was to measure the CSF concentration of cefepime in patients after neurosurgical operations, and to determine the penetration of the drug through an incomplete BBB.Methods Eight patients who received ventricular drainage (VD group) and 5 who underwent lumbar puncture drainage (LPD group) were enrolled into this study. Cefepime (2 g) was injected intravenously in 30 minutes after the neurosurgeries. The concentrations of cefepime in the CSF and plasma were measured by high-pressure liquid chromatography (HPLC) at different time points.Results The CSF concentrations of cefepime at different time points in the VD group were significantly higher than those in the LPD group (P＜0.05). In the VD group, the concentration of cefepime in CSF reached the peak ((22.54±14.06)μg/ml) at 1 to 2 hours after the injection, while in the LPD group at 4 hours ((5.61±3.73) μg/ml). In both groups, the peak was higher than the MIC90 of most common bacteria in intensive care unit. The ratio of CSF to plasma cefepime concentrations ranged from 0.30 to 2.14 in the VD group and 0.03 to 1.14 in the LPD group.Conclusion After neurosurgeries, CSF concentration of cefepime can reach a therapeutic level. Thus, the drug could be used to prevent and treat postoperative intracranial infection。

Abstract：Background Tissue Doppler imaging (TDI) has provided an objective means to quantify global and regional left ventricular (LV) and right ventricular (RV) function with improved accuracy and greater reproducibility than conventional echocardiography. This study was conducted to assess RV myocardial systolic activation by TDI in subjects with pulmonary arterial hypertension (PAH).Methods A total of 30 patients with PAH and 30 healthy volunteers, all comparable in age and sex, underwent standard Doppler echo and TDI. Using pulsed Doppler echocardiography combined with TDI, the following regional parameters were evaluated in three different myocardial segments (RV basal lateral wall, basal septal, and LV basal lateral) on apical 4-chamber view: systolic (Sm), early- and late-diastolic (Em and Am) peak velocities. RV myocardial systolic activation delay was defined as the difference in time to peak TDI systolic velocities between the RV basal lateral wall and basal septal. In addition, RV end-diastolic and end-systolic areas were measured to calculate RV fractional area change from the same apical 4-chamber view.Results Compared with the control group, patients with PAH showed increased RA and RV end-diastolic diameter (RA:(4.5±1.2)cm vs (3.0±0.8)cm, P＜0.05 and RV: (4.8±1.9)cm vs (3.4±0.5)cm, P＜0.05) and reduced RV fractional area change; (35±14)% vs (56±9)%, P＜0.05. These PAH patients showed lower myocardial peak velocities and a significant activation delay compared with controls (P＜0.05). Moreover, a strong correlation between RV myocardial systolic activation delay and RV fractional area change was shown in patients with pulmonary arterial hypertension (r = -0.82).Conclusions In PAH, RV myocardial systolic activation was markedly delayed, which was directly related to the RV fractional area change. RV myocardial systolic activation delay assessed by TDI could offer a unique approach to predict RV dysfunction。

Abstract：Background Folic acid is very important for embryonic development and dihydrofolate reductase is one of the key enzymes in the process of folic acid performing its biological function. Therefore, the dysfunction of dihydrofolate reductase can inhibit the function of folic acid and finally cause the developmental malformations. In this study, we observed the abnormal cardiac phenotypes in dihydrofolate reductase (DHFR) gene knock-down zebrafish embryos,investigated the effect of DHFR on the expression of heart and neural crest derivatives expressed transcript 2 (HAND2)and explored the possible mechanism of DHFR knock-down inducing zebrafish cardiac malformations.Methods Morpholino oligonucleotides were microinjected into fertilized eggs to knock down the functions of DHFR or HAND2. Full length of HAND2 mRNA which was transcribed in vitro was microinjected into fertilized eggs to overexpress HAND2. The cardiac morphologies, the heart rates and the ventricular shortening fraction were observed and recorded under the microscope at 48 hours post fertilization. Whole-mount in situ hybridization and real-time PCR were performed to detect HAND2 expression.Results DHFR or HAND2 knock-down caused the cardiac malformation in zebrafish. The expression of HAND2 was obviously reduced in DHFR knock-down embryos (P＜0.05). Microinjecting HAND2 mRNA into fertilized eggs can induce HAND2 overexpression. HAND2 overexpression rescued the cardiac malformation phenotypes of DHFR knock-down embryos.Conclusions DHFR plays a crucial role in cardiac development. The down-regulation of HAND2 caused by DHFR knock-down is the possible mechanism of DHFR knock-down inducing the cardiac malformation。

Abstract：Background Numerous studies have reported a relationship between hepatitis C virus (HCV) genotype and the response to interferon therapy. Despite high sensitivity and specificity, genotyping methods can be performed only on HCV RNA positive samples. Serotyping might be a rapid and cost effective method for determining HCV genotypes,especially in patients with previously undetectable HCV RNA. In this study, an enzyme linked immunosorbent assay (ELISA) method for HCV serotyping with the genotype specific, synthetic peptides derived from HCV nonstructural 5a (NS5A) region was developed.Methods Based on 45 sequences, representing HCV genotypes 1-6 from Genebank, we synthesised 305 overlapping 30-mer peptides within NS5A region at positions 2182-2343 of HCV. All peptides for antigenic reactivity were tested by enzyme immunoassay with 69 human sera with antiHCV positive representing genotype 1-6. Forty hepatitis C patient sera were serotyped using serotype specific, synthetic peptides and genotyped by sequencing analysis.Results The correspondence of amino acids in HCV NS5A region with amino acids in positions 2182-2343 was very low among different genotype peptides. The highly conserved sequences were residues 2182-2211(R1), 2272-2301 (R7)and 2302-2331 (R9): the highly variable 2212-2241 (R3) and 2257-2286 (R6). Using 305 peptides, antigenic regions were located in R3, R7 and R9. Eighteen peptides from highly conserved region representing genotypes 1 to 6 showed broad immunoreactivity with sera containing antibody to all HCV genotypes. Immunoreactivity of the peptides from highly variable region was stronger with similar genotype sera. Twelve unique peptides showed highly, genotype specific,reactivity with types 1 and 3 sera. Type 2 genotype specific peptides had cross reaction with type 3 serum. No type 4, 5 or 6 specific peptides were selected. The serotyping results showed high agreement with sequencing analysis.Conclusions The major antigenic regions in HCV NS5A region were at 2212-2241(R3), 2272-2301(R7) and 2302-2331(R9). Eighteen peptides from highly conserved region show genotype independent, immunoreactivity, useful for antiHCV antibody test. Twelve peptides from highly variable region show genotype 1 and 3 dependent immunoreactivity, useful for determining HCV serotype, especially for patients with previously undetectable HCV RNA。

Abstract：Background The public vaccination program of hepatitis B virus (HBV) was launched during 1984 in Taiwan, China.However, the long-lasting protective efficacy of HBV vaccination among adolescents older than 15 years of age was seldom recorded.Methods A seroepidemiological survey was conducted among 4575 first-year university students in Taiwan, China during 2000 to 2003, including the serological data of HBV by testing HBV surface antigen (HBsAg), surface antibody (anti-HBs), HBV core antibody (anti-HBc) and demographic information.Results HBsAg carrier rate among male university students born before the initiation of the HBV vaccination program decreased from 12.8% to 4.8% among those born after the vaccination program (P＜0.001, X2 test for linear trend).Similarly, HBsAg carrier rate among female university students born before the initiation of the HBV vaccination program decreased from 8.1% to 2.7% among those born after the vaccination program (P＜0.001, X2 test for linear trend). Both male and female students in eastern Taiwan had the highest HBsAg carrier rate compared with the other places. Using multiple logistic regression analysis, compared with students born after July 1984, the adjusted OR of HBsAg carrier rate decreased from 3.10 for students born before June 1981 to 1.56 for students born from July 1983 to June 1984 (95%CI 1.96-4.91, P＜0.001; 95%C/1.06-2.28, P=0.024; respectively).Conclusions Public vaccination provides long-lasting protection again HBV infection among the university students in Taiwan, China older than 18 years of age. There is a geographic variation of HBV infection among young adults in Taiwan,China。

Abstract：Background Obesity is now an epidemic in most parts of the world. In this cross sectional study, we report the most recent data on obesity in Hong Kong Chinese working population and compare the changes over 10 years.Methods Between July 2000 and March 2002, 5882 adult subjects from the working class in Hong Kong were recruited (2716 men (46.2%) and 3166 women (53.8%)). They were randomly selected using computer generated codes according to the distribution of occupational groups. Results of this study were compared with the data collected from a prevalence survey for cardiovascular risk factors in a Hong Kong Chinese working population conducted in 1990 (1513 subjects, 910 men (60.1%) and 603 women (39.9%)).Results Standardized percentages of overweight, obesity, and central obesity, in Hong Kong Chinese working population were 59.7%, 35.0%, 26.7% in men and 32.0%, 21.7%, 26.7% in women. Compared to the data collected in 1990, the percentage of obesity increased by 5% in men and reduced by 6% in women. The percentage of central obesity doubled in men (from 12.2% to 26.7%) but remained stable in women.Conclusions There is a doubling of the percentage of central obesity in Hong Kong Chinese working men over previous decade. Education and proper lifestyle modification program to tackle this social health issue are urgently indicated。

Background Human group O red blood cells have great benefit in specialized transfusion areas such as armed conflict and natural calamity. The group B antigen differs structurally from group O antigen only by the addition of one terminal α-linked galactose residue. In this study we aimed to remove the terminal galactose from group B red blood cell to get group O red blood cell.Methods α-galactosidase cDNA was cloned by RT-PCR from Catimor coffee beans grown on Hainan Island of China.The vector for α-galactosidase cDNA expression was constructed and transferred into Pichia pastoris cells by electroporation. The transgenic cells were cloned by fermentation and the recombinant α-galactosidase was purified by ion exchange chromatography. After studying the biochemical characters of α-galactosidase, we have used it in converting human erythrocytes from group B to group O.Results The purity of recombinant α-galactosidase was higher than 96%, which was thought to be suitable for the use of blood conversion. Enzymatically converted human group O red blood cells (ECHORBC) exhibited membrane integrity,metabolic integrity, normal cell deformation and morphology. There were no coagulation between ECHORBC and any group of human blood. The ECHORBC will keep normal structure and function for a period of 21 days at 4℃ in monoammoniumphosphate nutrient solution. Experiments with Rhesus monkeys and gibbons showed that transfusion of enzymatically converted erythrocytes was safe.Conclusion ECHORBC can be easily obtained from group B red blood cell by α-galactosidase digestion. This study suggests that ECHORBC could be transfused to patients safely and efficiently.

Abstract：Objective The results of studies on association between -148C/T polymorphism in promoter region of β-fibrinogen gene and susceptibility to cerebral infarction in Chinese population are controversial. In this study, we summarize the results of published works in this field by a meta-analysis.Data sources Genetic association studies evaluating the β-fibrinogen gene -148C/T polymorphisms and cerebral infarction involving Chinese population published before December 2005 were collected from PubMed, EMBASE and CNKI.Study selection Case control studies involving unrelated, Han subjects aged from 18 to 80 years, and the internationally recognized diagnostic standard of cerebral infarction and genotype frequencies in control group consistent with Hardy-Weinberg equilibrium were used. Publication bias was tested by funnel plot and the odds ratios of all studies were combined dependent on the result of heterogeneity test among the individual studies. The software Review Manager (Version 4.2) was used for meta-analysis.Results Eleven studies including 1223 patients and 1433 controls met the selection criteria. There was no heterogeneity among the odds ratios (ORs) of individual studies (x2=17.82, P=0.06). The combined OR of susceptibility to cerebral infarction in -148T allele carriers compared to the wild homozygote was 1.32 (95%C/1.12 to 1.55, P=0.0008).In the patients with cerebral infarction, the average plasma fibrinogen level of allele T carrier was 0.42 g/L (95%CI 0.29 to 0.54, P＜0.001), higher than that of -148C/C homozygous ones.Conclusions β-fibrinogen gene -148C/T polymorphism might contribute to susceptibility to cerebral infarction in Han Chinese. To reach a definitive conclusion, further gene to gene and gene to environment interactions studies on β-fibrinogen polymorphisms and cerebral infarction with large sample size are required。

Podocytes in renal glomerulus express unusual intermediate filament proteins (IFs) for visceral epithelial cells. IFs cytoskeleton is mainly composed of vimentin, nestin and desmin. Tissue injury is often accompanied by changes in gene expression of IFs.1 Enhanced desmin staining in podocytes are observed in a variety of rat experimental models of podocyte injury including puromycin aminonucleoside (PAN)nephrosis.2-4 It has not been elucidated whether expression of vimentin and nestin is up-regulated in podocyte injury. To further gain insight into expression of IFs in podocytes, we investigated the time course of vimentin, nestin and desmin in PAN nephrosis。

Multiple toes transplantation is an effective method for restoring the prehension of the hands.1-5 Generally, the thumb, index and middle finger are reconstructed by transferring a free big toe skin-nail flap with the second and third toes from a same foot. The great saphenous vein, superficial veins on the dorsum of the foot, the dorsalis pedis artery, and the first dorsal metatarsal artery are resected to be used as the donor pedicle. The arterial pedicle is freed up to the dorsal digital artery of the big and second toes. Both the second and third toes are supplied by the dorsal digital artery of the second toe on the tibial side. By using unilateral toe donation to reconstruct a digitless hand, we can preserve the normal form and function of the contralateral foot.However, because of the small length of the digital arteries between the big and second toes, the first web space is limited. Here we present our experiences on two cases of hand reconstruction with unilateral three toes transplantation. Good function and motion range of the thumb were achieved in both the patients。