[Multimodal imaging features of acute macular retinopathy in patients with COVID-19].

Objective: To investigate the multimodal imaging characteristics of acute macular retinopathy (AMR) and/or parafoveal acute middle maculopathy (PAMM) in patients with coronavirus disease 2019 (COVID-19). Methods: It was a cross-sectional study. Eight patients (15 eyes) diagnosed with AMN and/or PAMM, who presented for their initial visit at Kaifeng Eye Hospital between December 17 and December 31, 2022 and were also confirmed positive for COVID-19, were enrolled as the observation group. The patients were classified into four types based on swept-source optical coherence tomography (SS-OCT) findings. Fifteen healthy volunteers (15 eyes) without ocular or systemic diseases were recruited as the healthy control group, and one eye was randomly selected for analysis. All participants underwent detailed ophthalmic examinations, including best-corrected visual acuity (BCVA), slit-lamp biomicroscopy, fundus photography (FP), intraocular pressure measurement, fundus infrared imaging, OCT and OCT angiography (OCTA). The foveal avascular zone (FAZ) area of the macular center was measured. General information and multimodal imaging findings were collected and analyzed. The superficial capillary plexus vessel density (SCP-VD) and deep capillary plexus vessel density (DCP-VD) were measured in circular areas with diameters of 1.0 mm, >1.0 mm and ≤3.0 mm, and>3.0 mm and ≤6.0 mm centered on the foveal center, recorded as SCP-VD1.0, 3.0, 6.0 and DCP-VD1.0, 3.0, 6.0. Statistical analyses were performed using t-tests, Mann-Whitney U tests, and chi-square tests. Results: The observation group consisted of 6 males (11 eyes) and 2 females (4 eyes) with a mean age of (26.87±11.56) years. The healthy control group included 11 males (11 eyes) and 4 females (4 eyes) with a mean age of (28.75±12.30) years. There were no statistically significant differences in age and gender distribution between the two groups (all P>0.05). All patients in the observation group experienced high fever (≥39.0 ℃) and developed ocular symptoms during the febrile period or within 24 hours after fever resolution. Among all patients, there were 5 cases (7 eyes) of Type Ⅰ, 1 case (1 eye) of Type Ⅱ, 3 cases (4 eyes) of Type Ⅲ, and 2 cases (3 eyes) of Type Ⅳ. In Type Ⅲ and Ⅳ, 3 cases (4 eyes) exhibited weakly reflective cystic spaces in the outer plexiform or outer nuclear layers, and fundus photography revealed multiple gray or reddish-brown lesions in the macular region. One case (1 eye) showed retinal superficial hemorrhage. Cotton wool spots were observed in 2 cases (4 eyes). Fundus infrared imaging showed that Type Ⅰ manifested as weak reflectivity lesions in the parafoveal central zone, with the tip pointing towards the fovea. Type Ⅱ showed no apparent abnormalities in the macular region, while Type Ⅲ and Ⅳ displayed map-like weak reflective lesions spanning the foveal center. OCTA findings demonstrated that SCP-VD1.0 in the observation group was 6.93% (4.77%, 6.93%), significantly lower than the healthy control group's 10.66% (8.05%, 10.55%) (U=174.00, P=0.016). SCP-VD3.0 in the observation group was 37.14% (32.15%, 43.48%), also lower than the healthy control group's 43.06% (38.95%, 46.55%) (U=174.00, P=0.016). DCP-VD3.0 in the observation group was 48.20% (46.11%, 50.33%), lower than the healthy control group's 51.10% (50.04%, 53.02%) (U=188.00, P=0.009). DCP-VD6.0 in the observation group was 49.27% (47.26%, 51.67%), lower than the healthy control group's 52.43% (50.07%, 53.82%) (U=70.00, P=0.004). There were no significant differences in SCP-VD6.0 and DCP-VD1.0 between the two groups (both P>0.05). Conclusions: Acute macular retinopathy in patients with COVID-19 can involve all retinal layers and present as segmental hyper-reflectivity on SS-OCT. Fundus infrared imaging reveals weak reflectivity in the affected area, fundus photography shows multiple gray or reddish-brown lesions in the macular region, and OCTA demonstrates a decrease in SCP-VD and DCP-VD.

Transcriptomic differential lncRNA expression is involved in neuropathic pain in rat dorsal root ganglion after spared sciatic nerve injury

Dorsal root ganglia (DRG) neurons regenerate spontaneously after traumatic or surgical injury. Long noncoding RNAs (lncRNAs) are involved in various biological regulation processes. Conditions of lncRNAs in DRG neuron injury deserve to be further investigated. Transcriptomic analysis was performed by high-throughput Illumina HiSeq2500 sequencing to profile the differential genes in L4-L6 DRGs following rat sciatic nerve tying. A total of 1,228 genes were up-regulated and 1,415 down-regulated. By comparing to rat lncRNA database, 86 known and 26 novel lncRNA genes were found to be differential. The 86 known lncRNA genes modulated 866 target genes subject to gene ontology (GO) and KEGG enrichment analysis. The genes involved in the neurotransmitter status of neurons were downregulated and those involved in a neuronal regeneration were upregulated. Known lncRNA gene rno-Cntnap2 was downregulated. There were 13 credible GO terms for the rno-Cntnap2 gene, which had a putative function in cell component of voltage-gated potassium channel complex on the cell surface for neurites. In 26 novel lncRNA genes, 4 were related to 21 mRNA genes. A novel lncRNA gene AC111653.1 improved rno-Hypm synthesizing huntingtin during sciatic nerve regeneration. Real time qPCR results attested the down-regulation of rno-Cntnap lncRNA gene and the upregulation of AC111653.1 lncRNA gene. A total of 26 novel lncRNAs were found. Known lncRNA gene rno-Cntnap2 and novel lncRNA AC111653.1 were involved in neuropathic pain of DRGs after spared sciatic nerve injury. They contributed to peripheral nerve regeneration via the putative mechanisms.

Whole genome sequencing identified new somatic mutations for chronic myelomonocytic leukemia.

OBJECTIVE: We aimed to gain new insight into the molecular alterations of Chronic Myelomonocytic Leukemia (CMML). PATIENTS AND METHODS: We performed whole-genome sequencing (WGS) and subsequent Sanger sequencing validation analysis in three individuals with CMML. Genomic DNA samples from bone marrow and matching buccal mucosa samples were sequenced. RESULTS: For all six samples, a total of 806.43 Gb data were generated, achieving a minimum mean depth of 30.76. A total of 22 somatic variants were found to be protein-altering, including 1 exonic frame shift indel, 18 missense SNVs, 2 stop gain SNVs, and 1 stop loss SNV. We focused on the five novel variants which have not been reported in known databases and successfully validated three missense SNVs in AKAP4, COL2A1, and MAML1, respectively. CONCLUSIONS: WGS analyzes provided us a new insight into the molecular events governing the pathogenesis of CMML. The somatic variants we reported here may provide new targets for further therapeutic studies.

Establishment of cytochrome P450 3A4 and glutathione S-transferase A1-transfected human hepatoma cell line and functional analysis.

This study aimed to enhance the drug metabolism function of the human hepatoma cell line C3A and to explore the related significance for patients with severe liver disease. The important liver phase I and phase II drug metabolism enzymes, cytochrome P450 3A4 (CYP 3A4) and glutathione S-transferase A1 (GST A1), were constructed into a double expression vector and then transfected into C3A cells. Furthermore, in order to increase the expression of CYP 3A4 and GST A1, they were optimized according to human optimal codons. Another double-expression vector, pBudCE4.1-optimized CYP 3A4-optimized GST A1, was constructed and then transfected into C3A to establish a stable cell line. The drug metabolism function of C3A was evaluated. Sequence determination and analysis results showed that the recombinant plasmid pBudCE4.1-CYP 3A4-GST A1 met the application standard and its transfection was successful. The expression and activity of CYP 3A4 and GST A1 in unoptimized C3A cells were higher than those in blank C3A cells. Unoptimized C3A had a better drug metabolism function. Although some C3A cells transfected with pBudCE4.1-optimized CYP 3A4-optimized GST A1 survived, they grew slowly, and were therefore not applicable in clinical practice. Unoptimized C3A is superior to blank C3A in drug metabolism, and could be applied in the bioartificial liver support system as a new material.

Early versus delayed laparoscopic cholecystectomy for acute cholecystitis.

AIM: Laparoscopic cholecystectomy, currently the gold standard treatment for cholelithiasis, has been extended to treating acute cholecystitis as well. However, operation timing remains controversial. The aim of this retrospective study was to compare our data on the timing of surgery for early and delayed laparoscopic cholecystectomy for acute cholecystitis. METHODS: From January 1, 2006 to December 31, 2010, 508 laparoscopic cholecystectomy procedures were performed, 149 of which for acute cholecystitis: 122 operations were defined as early (performed within 72 hours of symptom onset) and 27 as delayed (72 hours to 9 days from symptom onset). RESULTS: There were no statistically significant differences in operating time, conversion or complications rates between early and delayed procedures. The total length of hospital stay was longer for patients who had undergone a delayed procedure. The success rates were similar irrespective of the surgeon's level of experience. CONCLUSION: Patients operated on for acute cholelithiasis between 72 hours and up to 9 days after symptom onset may benefit similarly as from an earlier operation. Delayed laparoscopic cholecystectomy for acute cholelithiasis is a feasible and safe procedure that compares favorably with early laparoscopic cholecystectomy.

Transillumination-assisted orotracheal intubation: a comparison of the Bonfils fibrescope and the lightwand (Trachlight).

BACKGROUND: Because the Bonfils fibrescope has a semi-rigid optical stylet and is similar in shape to a lightwand, we aimed to evaluate and compare the efficacy of transillumination-assisted orotracheal intubation with the Bonfils fibrescope and the Trachlight(TM) lightwand in patients with normal airways. METHODS: As a preliminary investigation to form a basis for later studies, therefore, we performed a randomized, single-blind study of 300 patients with normal airways to compare the efficiency of Trachlight and transillumination-assisted Bonfils orotracheal intubation in these patients. In both groups, orotracheal intubation was performed using a transillumination technique. The first attempt and overall success rates of tracheal intubation, the times required, and any untoward effects were recorded. RESULTS: Although the overall success rates were similar for Bonfils and Trachlight intubations (97.3% and 98.7%, respectively), tracheal intubation was successful on the first attempt in 87.3% of patients with the Bonfils fibrescope compared with 95.3% of patients with the Trachlight (P < 0.05). The mean intubation time for the first attempt was 15 ± 5 s with the Bonfils fibrescope and 9 ± 2 s with the Trachlight (P < 0.001). Patients intubated using the Bonfils fibrescope also experienced significantly more sore throat and hoarseness than those intubated using the Trachlight. CONCLUSIONS: For patients with normal airways, the Trachlight is superior for orotracheal intubation with respect to reliability, rapidity, and safety compared with the Bonfils fibrescope used with the transillumination technique.

Intraoral fixation of endotracheal tubes using a suture in edentulous patients undergoing maxillofacial surgery.

Securing an endotracheal tube in completely edentulous patients undergoing maxillofacial surgery can pose difficulties. In this report, a readily available and easy method of securing the endotracheal tube to gums of the teeth using the suture in such a circumstance is described. This technique has been used successfully in more than 100 patients at our institutions. Our experience suggests that it can provide reliable tube fixation and does not hinder surgical access.

Molecular and functional characterization of the cystic fibrosis transmembrane conductance regulator from the Australian common brushtail possum, Trichosurus vulpecula.

Unlike eutherian mammals, the colon of the Australian common brushtail possum, Trichosurus vulpecula, a metatherian mammal, is incapable of electrogenic Cl(-) secretion and has elevated levels of electrogenic Na(+) absorption, while the ileum secretes HCO (3) (-) rather than Cl(-). In eutherian mammals, the cystic fibrosis transmembrane conductance regulator (CFTR) is essential for both Cl(-) and HCO (3) (-) secretion and the regulation of Na(+) absorption. Therefore, we have sequenced possum (p)CFTR, described its distribution and characterized the properties of cloned pCFTR expressed in Fischer rat thyroid (FRT) cells. pCFTR (GenBank accession No. AY916796) has a 1,478 amino acid open reading frame, which has >90% identity with CFTR from other marsupials and >80% identity with non-rodent eutherian mammals. In pCFTR, there is a high level of conservation of the transmembrane and nucleotide binding domains although, with the exception of other marsupials, there is considerable divergence from other species in the R domain. FRT cells transfected with pCFTR express mature CFTR protein which functions as a small Cl(-) channel activated by cAMP-dependent phosphorylation. In whole-cell recordings it has a linear, time and voltage-independent conductance, with a selectivity sequence P(Br) > P(Cl) > P(I) > P(HCO)(3) >> P(Gluconate). pCFTR transcript is present in a range of epithelia, including the ileum and the colon. The presence of pCFTR in the ileum and its measured HCO (3) (-) permeability suggest that it may be involved in ileal HCO (3) (-) secretion. Why the possum colon does not secrete Cl(-) and has elevated electrogenic Na(+) absorption, despite the apparent expression of CFTR, remains to be determined.

Sustained and stable hematopoietic donor-recipient mixed chimerism after unrelated cord blood transplantation for adult patients with severe aplastic anemia.

We evaluated the engraftment of donor cells from unrelated cord blood into adult patients with severe aplastic anemia (SAA) and the outcome of allo-CBSCT (cord blood stem cell transplantation). Nine patients were conditioned with decreased dosage of immunosuppressive agents of CTX (60 mg/kg) and ALG (120 mg/kg). The prophylaxis of GVHD consisted of standard CsA and MTX. Patients have a media age of 25.3 yr (range: 15-37), and a median weight of 57.2 kg (range: 52.5-60) at the time of transplantation. Cord blood searches were all conducted at Guangzhou Cord Blood Bank. The engraftment state of the donor cells into recipients was confirmed by microsatellite DNA fingerprinting and fluorescent quantitative PCR analysis. Engrafted evidence has been found in seven patients involved by biomolecular analyses showing donor-recipient mixed chimerism post-transplant which was stable and persistent. After a median follow up of 32.2 months (range: 4-69), seven patients were alive and disease free. This study shows that durable donor-recipient stable mixed chimerism can be achieved by unrelated CBSCT in patients with SAA. Umbilical cord blood could be employed as a source of hematopoietic stem cell for adult transplantation.

Umbilical cord blood transplant for adult patients with severe aplastic anemia using anti-lymphocyte globulin and cyclophosphamide as conditioning therapy.

Allo-CBSCT (cord blood stem cell transplant) has been applied in six adult patients with severe aplastic anemia (SAA). Anti-lymphocyte globulin (ALG) 40 mg kg(-1) d(-1) x 3 days combined with cyclophosphamide (CTX) 20 mg kg(-1) d(-1) x 3 days constituted a lower intensive conditioning regimen. The prophylaxis of GVHD consisted of standard CsA and MTX. Patients are all male having a mean age of 26.5 years (range 22-38), and a median weight of 55.6 kg (range 52-60 kg). Cord blood searches were all conducted at Guangzhou Cord Blood Bank. Three of six patients in our study received one unit of cord blood in a procedure, whereas for another three patients, two units of cord blood (double units) were infused at the same time in a transplant protocol. The nine units of umbilical cord blood (UCB) infused contained 1.6-10.7 x 10(7) nucleated cells/kg body weight of the recipient after thawing. HLA antigens were identical in one unit, 1 antigen mismatched in seven, 2 antigens mismatched in 1. As of February 2003, after a median follow up of 20 months (range 7-50), four patients are alive and disease free. Five patients engrafted with molecular biology analyses showing donor-recipient mixed chimerism post transplant which is stable and persistent. One patient died of severe infection in the third month from transplant and another patient died in the early stage post transplant of serious aspergillus infection without evidence of engraftment.

Role of postsynaptic density protein-95 in the maintenance of peripheral nerve injury-induced neuropathic pain in rats.

Our previous work has demonstrated that postsynaptic density protein-95, a molecular scaffolding protein that binds and clusters N-methyl-D-aspartate receptors at neuronal synapses, plays an important role in the development of peripheral nerve injury-induced neuropathic pain. The current study further investigated the possible involvement of postsynaptic density protein-95 in the maintenance of neuropathic pain. Mechanical and thermal hyperalgesia were induced within 3 days and maintained for 15 days or longer after unilateral injury to the fifth lumbar spinal nerve. The rats injected intrathecally with postsynaptic density protein-95 antisense oligodeoxynucleotide every 24 h for 4 days from day 7 to day 10 post-surgery exhibited not only a marked decrease in spinal cord postsynaptic density protein-95 protein expression but also a significant reduction in mechanical and thermal hyperalgesia on day 11 post-surgery. The rats injected with sense oligodeoxynucleotide did not display these changes. However, in the rats without nerve injury, postsynaptic density protein-95 antisense oligodeoxynucleotide given intrathecally every 24 h for 4 days did not affect responses to mechanical and thermal stimulation. In addition, postsynaptic density protein-95 antisense oligodeoxynucleotide did not change locomotor activity of experimental animals. Our results indicate that the deficiency of postsynaptic density protein-95 protein in the spinal cord significantly attenuates nerve injury-induced mechanical and thermal hyperalgesia during both the development and maintenance of chronic neuropathic pain. These results suggest that postsynaptic density protein-95 might be involved in the central mechanisms of chronic neuropathic pain and provide a novel target for development of new pain therapies.

Frequent expression of new cancer/testis gene D40/AF15q14 in lung cancers of smokers.

We found a significant correlation between lung cancer in smokers and the expression of a human gene, D40, predominantly expressed in testis and cancers. In an attempt to clone a novel human gene, we screened a cDNA library derived from a human B cell line and obtained a cDNA clone that we refer to as D40. A search for public databases for sequence homologies showed that the D40 gene is identical to AF15q14. D40 mRNA is predominantly expressed in normal testis tissue. However, this gene is also expressed in various human tumour cell lines and primary tumours derived from various organs and tissues, such as lung cancer. We examined the relationship between D40 expression and clinico-pathological characteristics of tumours in primary lung cancer. D40 expression did not significantly correlate with either histological type or pathological tumour stage. However, D40 expression was observed more frequently in poorly differentiated tumours than in well or moderately differentiated ones. Furthermore, the incidence of D40 expression was significantly higher in tumours from patients who smoke than in those from non-smokers. D40/AF15q14 is the first gene in the cancer/testis family for which expression is related to the smoking habits of cancer patients.

[A study on pathological changes and the potential role of growth factors in the airway wall remodeling of COPD rat models].

OBJECTIVE: To study the pathological features of the smooth muscles and collagen in small airways of the COPD rat models and their roles in the airway obstruction, to evaluate the relationship between TGF-beta(1), EGF and bFGF and the airway wall remodeling. METHOD: Rat COPD model (model group) was established by intratracheal instillation of lipopolysaccharide (LPS 200 microgram/200 microL) twice and exposure to cigarette smoke daily. Drug intervention groups received either daily inhalation of budesonide, ipratropine or heparin respectively, starting on the 8 th day or TGF-beta(1) monoclonal antibody (TB21) 0.5 mg twice (6 th and 19 th day) via the tail veinous injection. At the end of four weeks, the thickness of the smooth muscles and collagen in bronchi and pulmonary arteriole wall were measured by means of image analyzer (CMIAS). Expression and localization of the 3 growth factors were observed in trachea, bronchi and lung tissues by immunohistochemistry and in situ hybridization. The levels of PC III, Ln and HA in the serum and BALF were determined by the RIA method. RESULTS: Significant thickening of the smooth muscles and collagen were found in the bronchi and pulmonary arterioles of the model group in comparison with those of the control group. There was significant decrease in the thickness of the collagen and smooth muscles in the small airways in TB21 group and heparin group. Statistically negative relationships were shown between the thickness of either smooth muscles or collagen in the small airways and FEV(0.3) (all P < 0.05). The levels of PC III, Ln and HA in COPD rat models were higher than those of control groups to varying extent. Expressions of TGF-beta(1), EGF and bFGF in the epithelial cells of bronchi, endothelial cells of pulmonary arterioles and in the macrophages of the model group were significantly higher than those of control group. The above mentioned parameters were reduced in different extent in drug intervention groups, in particular, the smooth muscles thickness in heparin group and the collagen thickness in TB21 group were significantly decreased compared to the model group. CONCLUSION: Thickening of smooth muscles and collagen in the bronchi constitutes the fundamental pathology of airway remodeling in the rat COPD model. The excessive depositions of ECM are important characteristics of COPD. TGF-beta(1), EGF and bFGF may play an important role in the airway wall as well as pulmonary arteriole remodeling. The intervention against TGF-beta(1) and long term inhalation of heparin may be of use in the inhibition of airway remodeling in COPD.

[Antigen sandwiched ELISA for detection of total HIV antibodies].

OBJECTIVE: To establish a sensitive antigen sandwiched ELISA(AS) for detection of the total antibodies to HIV-1/2. METHODS: Based on the gene sequence of HIV-1/2 type and its coded amino acid structure, 5 polypeptides were synthesized as coated antigens using solid-phase method. These polypeptides were labelled with horseradish peroxidase. And the total antibodies of HIV-1/2 were detected with the same method. RESULTS: These reagents were detected by three batches of HIV panel from The National Institute for the Control of Pharmaceutical and Biological Products (NICPBP). The results indicated that the corresponding rate was 100%. The variant coefficient rate was less than 10%. A comparison of antigen sandwiched ELISA with indirect ELISA in detection of a panel(20 positive sera and 20 negative sera) from the NICPBP showed that the general coincident rate of indirect ELISA was 92.5% and the sandwiched system was 100%. The HIV-AS diagnostic reagent kits have passed the quality examination of NICPBP. A comparison of antigen sandwiched ELISA with Yapei reagents in detection of 90 normal sera and 88 positive sera for HIV-1/2 showed that the coincident rate was 100%. The reagents were stable at 37 degrees C for 4 days. This indicated that our reagents were highly specific, sensitive and stable. CONCLUSION: Our antigen sandwiched ELISA reagent for total antibodies of HIV has a merit of sensitivity specificity and stability. It can be clinically used for detection of HIV-1/2 infection and in blood bank for the screening of blood donors.

Umbilical cord blood transplantation from unrelated HLA-matched donor in an adult with severe aplastic anemia.

A 23-year-old male suffering from severe aplastic anemia (SAA) weighing 60 kg was successfully treated by unrelated allo-CBSCT (cord blood stem cell transplantation). A six-loci HLA-identical umbilical cord blood (UCB) was infused after conditioning with low-dose cyclophosphamide (CTX) and antilymphocyte globulin (ALG). The prophylaxis of GVHD consisted of CsA and MTX. The infused cord blood provided 1.89 x 10(7) nucleated cells per kg, CD34-positive cells: 0.89%. Neutrophils >0.5 x 10(9)/l were reached 10 days after transplant, and platelets greater than 50.0 x 10(9)/l at day 26. RBC and platelet transfusion independence were reached on days 15 and 18. The patient developed grade 1 skin GVHD 10 months after engraftment of the donor cells. Microsatellite DNA fingerprinting indicated a stable and persistent donor-recipient mixed chimerism, whilst the circulating red cells remain of host origin.

Serological and histological findings in infection and transmission of GBV-C/HGV to macaques.

Seven healthy macaques were inoculated with the GBV-C/HGV-RNA serum from a non-A-E hepatitis patient. The serology and pathology of the liver in the animals were observed. The results indicated that all inoculated animals were infected with a GBV-C/HGV-RNA viremia and had mildly abnormal alanine transaminase levels during the infectious period. The histology, immuno-histochemistry, and in situ hybridization in the liver tissues of the inoculated animals also showed that there was a very mild hepatitis with the positive antigenic expression and the genome of GBV-C/HGV-NS5 in hepatocytes. The pathological changes in the infected animals appeared to become normal whether or not GBV-C/HGV-RNA viremia persisted. There is a possibility that the mild virulence of the GBV-C/HGV to the host became harmless with time after inoculation. Infection and the transmission of the GBV-C/HGV virus in the macaques provides an appropriate animal model and new information about GBV-C/HGV infection in both humans and animals. It is possible that this virus is a mild and self-limited pathogenic agent to the hepatic cells of primates.

[The potential role of growth factor in the airway wall remodeling of a chronic obstructive pulmonary disease rat model and the effects of drugs on them].

OBJECTIVE: To evaluate the expression and distribution of transforming growth factor-beta1 (TGF-beta1), epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) in the lung tissue of chronic obstructive pulmonary (COPD) rat models and the relationship between these growth factors and the airway wall remodeling. The effects of drugs on them were also investigated. METHODS: The COPD rat model (model group) was established by intratracheal instillation of lipopolysaccharide twice and daily exposure to cigarette smoking. Drug intervention groups received daily inhalation of heparin since the second week and TGF-beta1 monoclonal antibody (TB21) 0.5 mg twice through the tail veins. At the end of four weeks, the thickness of the smooth muscle and collagen in bronchi and pulmonary arterioles were measured by computer image analyzer, also the protein and gene relative content of these growth factors as well as the effects of drugs on them were observed. RESULTS: There was a significant increase in the smooth muscle and collagen thickness in the bronchi and pulmonary arterioles of the model group in comparison with that of the control group (P < 0.01), the relative contents for TGF-beta1, EGF and bFGF in the epithelial cells of the bronchi, endothelial cells of the pulmonary arterioles and alveolar macrophages of the model group were significantly higher than those of control group (P < 0.001 approximately 0.05). The relative content for TGF-beta1 in TB21 group was significantly lower than that of model group (P < 0.01). These were statistical positive relationships between the smooth muscle e thickness of bronchi and the relative contents for TGF-beta1, EGF and bFGF in the epithelial cells, between the collagen thickness of the bronchi and the relative content for TGF-beta1, between the smooth muscle thickness of the pulmonary arterioles and the relative content for TGF-beta1 and EGF in the endothelial cells (P < 0.05 approximately 0.01). CONCLUSION: TGF-beta1, EGF and bFGF may play an important role in the airway wall and pulmonary arteriole structure remodeling in COPD, the intervention against TGF-beta1 and long term inhalation of heparin mat be helpful for the inhibition of airway wall remodeling in human COPD and worth of further observation.

[Umbilical cord blood stem cells transplantation from an unrelated donor into an adult with severe aplastic anemia].

OBJECTIVE: To evaluate the availability of the treatment of adult severe aplastic anemia with unrelated allo-cord blood stem cell transplantation (CBSCT). METHODS: HLA-matched unrelated cord blood transplantation has been successfully performed for an adult severe aplastic anemia patient. A unit of cord blood provided by Guangzhou Cord Blood Bank containing 1.89 x 10(7)/kg mononucleated cells, 1.8 x 10(4)/kg CFU-GM and of CD(34) positive cells was 0.009. The patient was conditioned with CTX (60 mg/kg) and anti-lymphocyte globulin (ALG, 120 mg/kg). GVHD prophylaxis consisted of both MTX and CsA. The CsA had been given for 8 months. RESULTS: The lowest ANC was 0.6 x 10(9)/L post-transplantation. The patient achieved an ANC of greater than 0.5 x 10(9)/L at 10 days, and the platelet of greater than 50.0 x 10(9)/L at day 20 after transplantation. He developed Grade 1 GVHD in the tenth month after grafting. Microsatellite DNA fingerprinting indicated a stable donor-recipient mixed chimerism, whilst the circulating red cells remained host origin. CONCLUSION: It is the first report in China on successful treatment of adult severe aplastic anemia by unrelated allo-CBSCT.

Molecular analysis of the GCF gene identifies revisions to the cDNA and amino acid sequences(1).

GC factor (GCF) was reported as a transcriptional regulator that binds to a specific GC-rich sequence in the epidermal growth factor receptor (EGFR) gene promoter and represses its expression. In this paper, we present the data on three revisions of the cDNA sequence that lead to significant changes of the amino acid sequences of the published GCF. Firstly, 5'-rapid amplification of cDNA end (5'-RACE) analysis revealed that the 308 nucleotides of 5'-end of the previously published GCF cDNA does not exist at the 5'-end of the RACE product. Simultaneously, the correct 5'-end cDNA sequence of 31 nucleotides was identified. Secondly, the 'T' at the position 787 of the published GCF cDNA was not observed. Finally, a new sequence of 114 nucleotides was identified between the positions 851 and 852 of the published cDNA sequence. The revisions result in a GCF cDNA of 2661 nucleotides that encodes a protein of 781 amino acids, replacing the highly basic region of the amino-terminus of the published GCF with a new sequence of 147 amino acids. In this era of massive gene cloning and sequencing, this study is a warning to the biological research of recent years.

[Revisions of the cDNA and primary protein structure of human transcription factor GCF].

GC factor (GCF) was reported as a transcriptional regulator that binds to specific GC-rich sequences in the epidermal growth factor receptor (EGFR) gene promotor, repressing its transcription (Kageyama R. and Pastan I. Cell, 59: 815-825, 1989). In this paper, the author presents revisions of the cDNA and the amino acid sequences of the GCF. 1) 5' rapid amplification of cDNA end (5'RACE) for analysis of RNA of a cancer cell line, A431, was performed, which revealed that the 5' end of GCF cDNA was fused to a 308 bp fragment of other cDNA; simultaneously, the real 5' end cDNA sequence with 31 bp was identified. RNase protection assay presented a main protected band, which was consistent with the result of the RACE analysis. 2) T at the position 787 of the previously reported GCF cDNA was absent from RT-PCR on A431 total RNA. 3) A new sequence with 114 bp was observed on A431 RNA between the positions 851 and 852 of the already reported cDNA by RT-PCR. These observations were confirmed by RT-PCR analyses of RNAs prepared from several other human cell lines, including a non-transformed one (HFL), and white blood cells derived from a normal person. 4) Sequence of genomic GCF DNA was consistent with the new cDNA sequence but not with the previously reported one. 5) The remaining sequence of GCF cDNA was found to be identical to that of the previously reported GCF, based on the results of RT-PCR analyses of RNA prepared from human white blood cells. 6) By the corrections, the GCF cDNA consisted of 2661 bp nucleotides. This revised GCF cDNA (the wild type) encodes a protein of 781 amino acids, including two new sequence regions of 186 amino acids on the N-terminal side of this protein. The revisions eliminated the highly basic region of the amino-terminus of the previously reported GCF, while the other three fourth amino acid sequences of the GCF protein that contained leucine-zipper-like domain had not changed. The revised GCF had no highly homologous protein in the database except the previously reported GCF. 7) The author has developed a specific antibody to human GCF protein. This antibody specifically recognized a protein with a molecular weight of approximately 100 kDa present in the extracts from human cell lines, as confirmed by immunoprecipitation followed by Western blotting. 8) Indirect immunofluorescence of A431 and HeLa cells using the anti-GCF antibody showed that the GCF protein was localized in the nucleus, suggesting that the revised GCF is a nucleoprotein.