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Extracorporeal carbon dioxide removal (ECCO 2 R) is a respiratory support technique based on extra-pulmonary gas exchange, which can effectively remove carbon dioxide generated in-vivo, reducing the requirements of respiratory support from mechanical ventilation. With improvements in extracorporeal life support technologies and increasing clinical experience, ECCO 2 R has potential value in clinical application with acute respiratory distress syndrome (ARDS). This review article discusses the principles of ECCO 2 R, its relevant indications for ARDS, clinical evidence, existing issues, and future directions, aiming to provide more references for the application in ARDS.

Objective: To explore the application value of flexible endoscopy and rigid endoscopy in the clinical examination of chronic sinus tract wounds with different shapes. Methods: A retrospective observational study was conducted. From January 1 to December 23, 2019, a total of 46 patients with chronic sinus tract wounds, who met the inclusion criteria were admitted to the Wound Healing Center of Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, including 23 males and 23 females, aged 18-81 (48±21) years. On admission, computer tomography (CT) imaging and three-dimensional reconstruction were performed to examine the shapes of wound sinus tract and classify the wounds, with the lengths of wound sinus tract by CT imaging examination (hereinafter referred to as reference lengths) recorded. The lengths of wound sinus tract were examined and measured by rigid endoscopy and flexible endoscopy. The wounds with and without obviously curved sinus tract were classified into curve group and linear group respectively, and the deviation rates between the lengths of wound sinus tract measured by flexible endoscopy or rigid endoscopy and the reference lengths (hereinafter referred to as deviation rates of lengths) in each group were calculated. The difference between the deviation rates of lengths examined by flexible endoscopy and rigid endoscopy and the differences between the above two and the deviation rate of reference lengths (0) in each group were compared. Data were statistically analyzed with paired sample t test and Wilcoxon signed rank sum test. Results: CT imaging and three-dimensional reconstruction showed that there were 4 types of wound sinus tract, including tubular (36/46), lamellar (4/46), club-mallet (4/46), and irregular (2/46) shape. Tubular wounds were further divided into type I (23/36), type L (4/36), and type Y (9/36). Wounds with type I tubular, lamellar, and club-mallet sinus tract were classified into linear group (31/46), while those with type Y tubular, type L tubular, and irregular sinus tract were classified into curve group (15/46). In linear group, the deviation rates of lengths examined and measured by rigid endoscopy and flexible endoscopy were 0. In curve group, the deviation rate of lengths examined and measured by flexible endoscopy was 0 (0, 0.58%), which was significantly lower than 41.18% (31.68%, 48.41%) examined and measured by rigid endoscopy, Z =-3.408, P <0.01; the deviation rate of lengths examined and measured by rigid endoscopy (40±19)% was significantly higher than the deviation rate of reference lengths ( t =8.343, P <0.01), while the deviation rate of the lengths examined and measured by flexible endoscopy was similar to the deviation rate of reference lengths ( Z =-1.342, P >0.05). Conclusions: Compared with rigid endoscopy, flexible endoscopy can observe the internal characteristics of chronic sinus tract wounds in a wider range in the clinical examination of this kind of wound, especially for the exploration of curved chronic sinus tract wounds. The promotion of this method will be conducive to the diagnosis and treatment of chronic sinus tract wounds.

According to a document issued by the General Office of National Health Commission, "one person, one diagnosis, and one room" is required in the process of outpatient consultation. However, the patient will need to go to another room for dressing change after the doctor checks the wound if sticking to the conventional layout of current wound repair specialist outpatient clinic in hospitals and following the regulation of "separation of diagnosis and treatment". To allow a patient walking back and forth with the exposed wounds to different clinics or going to another clinic for dressing change with the original dressing reapplied to the wound is against the regulation of nosocomial infection control and the principle of sterility. To ensure that the layout of the outpatient clinic in the wound repair outpatient department not only conforms to the principle of "one person, one diagnosis, and one room", but also meets the characteristics of the diagnosis and treatment process of chronic wounds, this paper proposes the layout of "large space and small partition" in the wound repair clinic.

Statistics show that 76.74% (4 688) of 6 109 patients with chronic wounds are over 50 years of age; the proportion of patients with underlying diseases in all age groups above 50 years ranges from 78.25% to 100.00%. Among the underlying diseases of chronic wound patients, the top four diseases are diabetes mellitus, cardiovascular and cerebrovascular diseases, hypertension, and respiratory diseases. The above underlying diseases and ages of patients are the susceptibility factors of coronavirus disease 2019 released by National Health Commission of China. It is an unavoidable fact that the patients with chronic wounds have to go to the hospital for treatment prescribed by the physician. At the same time, we found that quite a few patients preferred going far afield in choosing hospital for treatment due to various reasons. During the prevention and control of coronavirus epidemic, this " go far afield" style of seeking medical treatment may increase the exposure risk during travelling. Accordingly, we convened 36 wound care clinics in different regions in Shanghai to implement the " Five Measures" encouraging the patients with chronic wounds to seek medical treatment nearby. The principle of this operation is that when seeking medical treatment, patients with chronic wounds should try their best to reduce the travel distance as much as possible to minimize the exposure risk during the epidemic period, which will in turn support the campaign of epidemic prevention and control.

The " exploration of treatment technology of chronic wound with sinus tract supported by endoscope and related auxiliary methods" study had been conducted by Wound Healing Center of Ruijin Hospital, Shanghai Jiao Tong University School of Medicine. The feasibility and effectiveness of this technique have been confirmed, and good clinic results have been achieved. In order to further promote the new technology and its related research, the theoretical knowledge and technical experience accumulated in the early stage are summarized as diagnosis and treatment standard for treatment with endoscopy technique in chronic wounds with sinus tract, including four parts: the applicable subjects, the diagnosis and treatment procedures and classification criteria, the healing criteria, and the risk assessment and prevention measures. The purpose of this standard is to facilitate the application of standardized endoscopy technique, to make the most of its technological advantages, prevent risks, and provide a reference for the official version of the diagnosis and treatment standard.

Objective: To investigate whether adipose-derived stem cells (ASCs) from allogeneic diabetic rats can promote wound healing in diabetic rats or not and the mechanism. Methods: (1) Fifty-six male Wistar rats aged 12-16 weeks were divided into diabetic group and healthy group according to the random number table (the same grouping method below), with 28 rats in each group. Rats in healthy group were not treated with any treatment. Rats in diabetic group were injected with 10 g/L streptozotocin 60 mg/kg intraperitoneally in one time to establish the diabetic model. Four rats in diabetic group and 4 rats in healthy group were selected according to the random number table, and the adipose tissue in the inguinal region was taken to culture and purify ASCs, so as to obtain healthy rat-derived ASCs (hereinafter referred to as nASCs) and diabetic rat-derived ASCs (hereinafter referred to as dASCs). The third passage of nASCs ( n =3) and dASCs ( n =3) were taken, and the positive expression rates of cell surface differentiation antigens CD105, CD31, CD34, and CD44 were detected with flow cytometer for defining ASCs purity. (2) The rest 24 rats in healthy group and 24 rats in diabetic group were used to make three round full-thickness skin defect wounds with a diameter of 12 mm on the back of each rat. Immediately after injury, phosphate buffer saline (PBS), nASCs of 2×10(7)/mL, and dASCs of 2×10(7)/mL each in the volume of 0.5 mL were subcutaneously injected into three wounds and their margins of each rat, respectively. On post injury day (PID) 1, 3, 7, and 12, 6 rats in each group were selected according to the random number table to calculate the wound area, and the wound tissue was stained with hematoxylin-eosin to observe the histological morphology of the wound. (3) Human ASCs (hASCs) were subcultured, and the 4th to 7th passage of cells were used for the subsequent experiments. The hASCs were divided into 7 groups, with 12 samples in each group. Cells in blank control group were cultured with mesenchymal stem cell culture medium, and cells in simple advanced glycation end products (AGEs) group, simple protein group, simple high glucose group, simple high osmotic pressure group, AGEs-high glucose combination group, and protein-high osmotic pressure combination group were cultured with mesenchymal stem cell culture medium containing a final mass concentration of 100 mg/L AGEs, 100 mg/L bovine serum albumin (BSA), 28 mmol/L D-glucose, 28 mmol/L mannitol, 100 mg/L AGEs+ 28 mmol/L D-glucose, 100 mg/L BSA+ 28 mmol/L mannitol, respectively. Cell proliferation was detected by cell counting kit 8 at post culture hour (PCH) 2 and on post culture day (PCD) 2, 4 and 6. (4) The hASCs were divided into blank control group, simple AGE group, simple high glucose group, and AGE-high glucose combination group, with 12 samples in each group, which were treated the same as corresponding groups in experiment (3). On PCD 0, 2, 4, and 6, the positive expression rates of cell surface differentiation antigens CD105, CD44, and CD45 were detected by flow cytometer to estimate their homeostasis. (5) The hASCs were divided into AGE-high glucose combination group and protein-high osmotic pressure combination group, with 9 samples in each group, which were treated the same as corresponding groups in experiment (3). On PCD 2, 4, and 6, the expression of intracellular protein was detected by cyanine 3-streptavidin double-antibody sandwich technique. Data were processed with analysis of variance for factorial design, least significant difference test, and Bonferroni correction. Results: (1) The positive expression rates of CD44 in nASCs and dASCs were both higher than 96%, the positive expression rates of CD31 and CD34 were low, and the positive expression rates of CD105 were about 40%, which basically met the purity requirements. (2) The areas of wounds treated by three methods in rats of healthy group and diabetic group were similar on PID 1 ( P >0.05). In healthy group, compared with (0.682 1±0.078 9), (0.314 3±0.113 7), and (0.064 3±0.002 1) cm(2) of the PBS-treated wounds in rats, the area of nASCs-treated wounds in rats decreased significantly on PID 3, 7, and 12 [(0.464 1±0.092 6), (0.223 9±0.072 7), and (0.034 3±0.012 5) cm(2), P <0.05], the area of dASCs-treated wounds in rats decreased significantly on PID 3 and 12 [(0.514 1±0.124 1) and (0.043 7±0.032 8) cm(2), P <0.05] but was not obviously changed on PID 7 [(0.274 2±0.062 5) cm(2), P >0.05]. Compared with those of the dASCs-treated wounds of rats within the same group, the area of the nASCs-treated wounds of rats in healthy group decreased significantly on PID 3 and 7 ( P <0.05) but was not obviously changed on PID 12 ( P >0.05). In diabetic group, compared with (0.853 5±0.204 8), (0.670 5±0.164 8), and (0.131 4±0.074 4) cm(2) of the PBS-treated wounds in rats, the area of nASCs-treated wounds in rats decreased significantly on PID 3, 7, and 12 [(0.633 4±0.132 5), (0.331 8±0.023 5), and (0.074 2±0.003 8) cm(2), P <0.05], the area of dASCs-treated wounds in rats decreased significantly on PID 3 [(0.773 6±0.182 2) cm(2), P <0.05] but was not obviously changed on PID 7 and 12 [(0.510 6±0.192 2) and (0.114 4±0.003 1) cm(2), P >0.05]. Compared with the dASCs-treated wounds of rats within the same group, the area of the nASCs-treated wounds of rats in diabetic group was not obviously changed on PID 3 and 7 ( P >0.05) but decreased significantly on PID 12 ( P <0.05). There was no obvious difference in histological morphology of the wounds treated with three methods in rats of each group on PID 1. On PID 3, a small amount of microvessels were formed in the wounds treated with nASCs and dASCs of rats in both groups, but microvessel formation was almost undetected in the PBS-treated wounds. On PID 7, more small blood vessels and fibroblasts (Fbs) were observed in the wounds treated with nASCs and dASCs of rats in both groups, but the small blood vessels and Fbs were slightly less in the PBS-treated wounds. On PID 12, the wounds treated with nASCs and dASCs of rats in the two groups were covered by epithelial tissue, the granulation tissue in the PBS-treated wounds of rats in healthy group was not obvious, and the PBS-treated wounds of rats in diabetic group were not completely epithelialized. (3) Compared with those of blank control group, the cell number of hASCs in simple AGEs group decreased significantly on PCD 2, 4, and 6 ( P <0.05), which increased significantly on PCD 2 and 4 in simple high glucose group ( P <0.05), and that in AGEs-high glucose combination group decreased significantly on PCD 4 and 6 ( P <0.05). (4) Compared with that on PCD 4 within the same group, the positive expression rate of CD105 in hASCs decreased significantly in blank control group, simple AGEs group, and AGEs-high glucose combination group on PCD 6 ( P <0.05). The positive expression rate of CD44 was higher than 95%, and that of CD45 was less than 2% in hASCs of each group at each time point. (5) Detection values of 7 proteins were located in the confidence interval. The expression levels of basic fibroblast growth factor and tissue inhibitor of metalloproteinase-1 in hASCs of AGEs-high glucose combination group and protein-high osmotic pressure combination group showed increasing trend with the prolongation of culture time. The expression level of human monocyte chemoattractant protein 1 (MCP-1) in hASCs of AGEs-high glucose combination group showed increasing trend with the prolongation of culture time, while the expression level of growth-regulated oncogene (GRO) on PCD 6 was significantly higher than that on PCD 4 within the same group ( P <0.05); the expression levels of MCP-1 and GRO in hASCs of protein-high osmotic pressure combination group showed decreasing trend with the prolongation of culture time. The expression level of follistatin in hASCs of protein-high osmotic pressure combination group decreased obviously on PCD 4, while that in hASCs of AGEs-high glucose combination group was significantly lower on PCD 6 than that on PCD 4 ( P <0.05). The expression level of vascular endothelial growth factor (VEGF) in hASCs of protein-high osmotic pressure combination group decreased gradually with the prolongation of culture time, while that in hASCs of AGEs-high glucose combination group on PCD 4 decreased significantly as compared with that on PCD 2 ( P <0.05). The expression level of urokinase-type plasminogen activator receptor in hASCs of protein-high osmotic pressure combination group on PCD 6 was significantly higher than that on PCD 4 within the same group ( P <0.05) and that of AGEs-high glucose combination group on PCD 6 ( P <0.05). Conclusions: Both nASCs and dASCs can promote wound healing in rats with simple defect injury, but dASCs have no significant effect on wound healing in rats with diabetes mellitus, which may be related to the inhibition of ASCs proliferation and the influence of high glucose and AGEs intervention on their homeostasis and secretory function.

Objective: To explore the advantages of endoscopy combined with contrast fistulography in the clinical diagnosis and treatment of chronic wound with sinus tract adjacent to body cavity. Methods: Thirty-two patients (14 males and 18 females, aged 17 to 87 years) of chronic wounds with sinus tracts adjacent to body cavity, who underwent endoscopy combined with contrast fistulography (CT or magnetic resonance imaging) for the diagnosis and treatment in the Outpatient Department of Wound Healing Center of our hospital from October 2017 to March 2019, were enrolled in the study. Their diagnosis and treatment results were retrospectively analyzed. The following data were calculated. (1) The incidence rates of sinus wound involving body cavity or fistula. (2) The detection rates of sinus wound involving body cavity detected by routine examination and by endoscopy combined with contrast fistulography. (3) The detection rate of pathological features at deep part of wound by routine examination and by endoscopy combined with contrast fistulography. (4) The proportion of patients who benefited from routine examination and from endoscopy combined with contrast fistulography. Data were processed with paired chi-square test and Fisher's exact probability test. Results: (1) The incidence rate of sinus wound involving body cavity was 43.75% (14/32); the incidence rate of fistula was 0. (2) The detection rate of sinus wound involving body cavity detected by endoscopy combined with contrast fistulography was 43.75% (14/32), which was obviously higher than that by routine examination [12.50% (4/32), χ (2)=32.0, P <0.01]. (3) The detection rate of pathological features at deep part of wound by endoscopy combined with contrast fistulography was 37.50% (12/32), which was obviously higher than that by routine examination (0, P <0.01). (4) The proportion of patients who benefited from endoscopy combined with contrast fistulography was 71.43% (20/28), which was obviously higher than that from routine examination [12.50% (4/32), χ (2)=21.6, P <0.01]. Conclusions: Compared with routine examination, endoscopy combined with contrast fistulography is more accurate in detecting chronic wound with sinus tract adjacent to body cavity. The diagnosis and treatment of chronic wound with sinus tract adjacent to the body cavity can benefit from this joint examination.

The correct thoughts and principles of diagnosis and treatment of chronic refractory wounds need to be formulated. Through the relevant domestic and international consensus and based on clinical experience, the Thoughts and principles of diagnosis and treatment of chronic refractory wounds in China is proposed. It is considered that in the diagnosis and treatment of chronic refractory wounds, in the case of fully understanding the patient's medical history, the following thoughts and principles should be complied in order. (1) Pay attention to the cleanliness of the wound after being cleaned. (2) Reasonably perform debridement to avoid being " excessive" or " not thorough". (3) Reasonably perform examination, diagnosis, and differential diagnosis of pathogenic factors. (4) Treat according to etiology. (5) Find comorbidities and prevent adverse outcomes. (6) Select the correct wound treatment method reasonably and timely. When the conservative wound care treatment is considered, pay attention to embodying the concept of etiological treatment, treat the wound according to the principles of safety, phase, selectivity, and effectiveness, and make a reasonable choice of continuing conservative treatment or surgical treatment in time after completing the preparation of the wound bed. When surgical treatment is considered, pay attention to the selection of reasonable surgical method and donor site, pay attention to the healing rate of surgical wound site and the outcome of donor site, and give reasonable protection to the wound site after surgery. (7) Carry out rehabilitation treatment after wound healing and related health education.

Objective: To explore the application value of endoscope in probing the chronic wound with sinus tract in clinic. Methods: Twenty-eight chronic wounds with sinus tracts from 27 patients conforming to the inclusion criteria admitted to Outpatient Department of Wound Healing Center of Ruijin Hospital from December 2017 to March 2018 were investigated in a prospective and self-controlled trial. After being cleaned, the diameter of the opening of sinus tract was measured with a rule. A probe was used to measure the depth of a sinus tract according to the touch from the probe extremity in operation, and to measure the depth of a sinus tract that could be observed with naked eyes with the help of a pair of hemostatic forceps. Five minutes later, a probe was inserted deeply into the sinus tract to measure the depth under the endoscopic view combined with touch from the probe extremity in operation. Afterwards, the sinus tract was observed with endoscope, and the depth of the tract which could be observed under the endoscopic view was measured using a probe inserted deeply into the sinus tract. After completion of the above exploration, the sinus tract was infused with contrast agent Omnipaque 350 and scanned by computed tomography (CT) later to obtain its depth. The following indicators were calculated: the ratio of the depth of the sinus tract measured by CT to the diameter of the opening of the sinus tract (hereinafter referred to as the depth/diameter ratio of the sinus tract), the deviation rate comparing the depth of the sinus tract measured by conventional method (measured by probe only) and by endoscope (measured by probe under the endoscope view) with the depth of the sinus tract measured by CT (hereinafter referred to as the deviation rate of the measured depth of the sinus tract), the deviation rate comparing the depth of the sinus tract that could be observed measured by conventional method and by endoscope with the depth of the sinus tract measured by CT (hereinafter referred to as the deviation rate of the depth of the sinus tract that could be observed). Data were processed with paired t test. Pearson correlation analysis was applied to analyze the correlation between the depth/diameter ratio of the sinus tract and the deviation rate of the measured depth of the sinus tract and the deviation rate of the depth of the sinus tract that could be observed by conventional method and by endoscope. Results: The depth/diameter ratio of the sinus tract of this group of wounds was 1-32 (8±7). The deviation rate of the measured depth of the sinus tract and the deviation rate of the depth of the sinus tract that could be observed by conventional method were (19±14)% and (79±18)%, respectively, both obviously larger than (9±9)% and (25±25)% by endoscope ( t =3.837, 13.626, P <0.01). Positive correlation existed between the depth/diameter ratio of the sinus tract and the deviation rate of the measured depth of the sinus tract by conventional method, and between the depth/diameter ratio of the sinus tract and the deviation rate of the depth of the sinus tract that could be observed by conventional method and by endoscope ( r =0.514, 0.585, 0.651, P <0.01). However, there was no obvious correlation between the depth/diameter ratio of the sinus tract and the deviation rate of the measured depth of the sinus tract by endoscope ( r =0.113, P >0.05). Conclusions: Compared with the conventional method, application of endoscope is able to get more accurate data of chronic wounds with sinus tracts and observe the wounds with wider range.

Objective: To investigate the effects of denatured collagen type Ⅰ on differentiation of human fibroblasts into myofibroblasts. Methods: A small amount of normal skin donated by burn patients undergoing scar surgery was collected. Human fibroblasts were obtained by method of explant culture and then sub-cultured. The fourth passage of cells were used in the following experiments. (1) Fibroblasts were divided into normal collagen group and denatured collagen group according to the random number table, with 10 wells in each group. Fibroblasts in normal collagen group were cultured on normal collagen type Ⅰ coated coverslips. Fibroblasts in denatured collagen group were cultured on denatured type Ⅰ collagen coated coverslips. Expression of proliferating cell nuclear antigen (PCNA) was detected by immunohistochemical method, and the percentage of PCNA positive cells was calculated. (2) Another batch of fibroblasts were grouped and treated as in (1), with 12 wells in each group. Proliferation activity of cells was determined with methyl-thiazolyl-tetrazolium colorimetry method. (3) Another batch of fibroblasts were grouped and treated as in (1), and the microfilament morphology of cells was observed by rhodamine-phalloidin staining. (4) Another batch of fibroblasts were grouped and treated as in (1). Expression of α smooth muscle actin (α-SMA) of cells was detected by immunohistochemical method, and expression of OB-cadherin of cells was detected by immunofluorescence method. (5) Another batch of fibroblasts were divided into normal collagen, denatured collagen, and common coverslips groups according to the random number table, with 6 wells in each group. Fibroblasts in normal collagen and denatured collagen groups were treated as in (1), while fibroblasts in common coverslips group were cultured on coverslips without collagen coating. Expressions of α-SMA and OB-cadherin of cells were determined with Western blotting. (6) Another batch of fibroblasts were grouped and treated as in (5), and then the mRNA expressions of collagen type Ⅰ, collagen type Ⅲ, and α-SMA of cells were determined with real-time fluorescent quantitative reverse transcription polymerase chain reaction. Data were processed with t test, one way analysis of variance, and least-significant difference test. Results: (1) The percentage of PCNA positive cells in denatured collagen group was (83±9)%, significantly higher than (29±9)% in normal collagen group ( t =13.53, P <0.01). (2) The proliferation activity of fibroblasts in denatured collagen group was 0.32±0.06, significantly higher than 0.25±0.05 in normal collagen group ( t =3.06, P <0.01). (3) The microfilament of fibroblasts in normal collagen group was arranged vertically and in parallel way, paralleling the long axis of cells. The microfilament of fibroblasts in denatured collagen group was denser and thicker. (4) Most fibroblasts in normal collagen group showed long shuttle-like shape typically. Morphology of fibroblasts in denatured collagen group changed, and cells were obviously spreading. Expressions of α-SMA and OB-cadherin of fibroblasts in denatured collagen group were stronger than those in normal collagen group. (5) Expressions of α-SMA of fibroblasts in denatured collagen, normal collagen, and common coverslips groups were respectively 1.69±0.41, 0.89±0.27, and 1.46±0.42. Expression of α-SMA of fibroblasts in denatured collagen group was significantly higher than that in normal collagen group ( P <0.01). Expressions of OB-cadherin of fibroblasts in denatured collagen, normal collagen, and common coverslips groups were respectively 5.17±0.28, 2.21±0.10, and 4.01±0.56. Expression of OB-cadherin of fibroblasts in denatured group was significantly higher than that in normal collagen group ( P <0.01). (6) There was no significant difference in mRNA expression of collagen type Ⅰ of fibroblasts in denatured collagen, normal collagen, and common coverslips groups ( F =2.71, P >0.05). The mRNA expressions of collagen type Ⅲ and α-SMA of fibroblasts in normal collagen group were significantly lower than those in denatured collagen group ( P <0.01). Conclusions: Denatured collagen type Ⅰ may influence the activity of fibroblasts, thus inducing fibroblasts differentiating into myofibroblasts.

Objective: To simulate the environmental factors during the process of formation and evolution of hypertrophic scar, so as to explore the effects of moderate and severe hypoxia and low concentration of serum protein on the function of human hypertrophic scar fibroblasts (HSFs). Methods: Human HSFs were routinely cultured. Cells of the 3rd to the 6th passage were divided into 10.0% oxygen+ 10.0% fetal calf serum (FCS), 5.0% oxygen+ 5.0% FCS, and 0.5% oxygen+ 0.5% FCS groups according to the random number table. After being cultured with DMEM nutrient solution with no FCS for 24 h, the cells were cultured with the corresponding volume fraction of oxygen and FCS. Cell proliferation activity was determined with methyl-thiazole-tetrazolium assay (denoted as actual cell number). Content of total collagen was detected with Sirius red staining method (denoted as absorbance value). Protein expression levels of hypoxia-inducible factor 1α (HIF-1α), vascular endothelial growth factor (VEGF), transforming growth factor β 1 (TGF-β 1 ), B-cell lymphoma 2 (Bcl-2), and P53 were determined with Western blotting (denoted as ratio of gray value). Cell apoptosis rate was detected by in situ end labeling method. The sample numbers of each group in the above experiments were all 3. Data were processed with Kruskal-Wallis test and Dunnett test. Results: (1) Compared with 11 000±1 306 in 10.0% oxygen+ 10.0% FCS group, the cell proliferation activity was higher in 5.0% oxygen+ 5.0% FCS group (13 290±1 500, P <0.05), but lower in 0.5% oxygen+ 0.5% FCS group (6 999±765, P <0.05). (2) Compared with 0.039 6±0.004 2 in 10.0% oxygen+ 10.0% FCS group, the content of total collagen of cells was higher in 5.0% oxygen+ 5.0% FCS group (0.051 6±0.005 1, P <0.05), but lower in 0.5% oxygen+ 0.5% FCS group (0.015 6±0.002 4, P <0.05). (3) Compared with those in 10.0% oxygen+ 10.0% FCS group, the protein expression levels of HIF-1α, VEGF, TGF-β 1 , and Bcl-2 were increased (with P values below 0.05), with no obvious difference in protein expression level of P53 in 5.0% oxygen+ 5.0% FCS group ( P >0.05), whereas the protein expression levels of HIF-1α, VEGF, TGF-β 1 , and Bcl-2 were decreased (with P values below 0.05), while the protein expression level of P53 was increased in 0.5% oxygen+ 0.5% FCS group ( P <0.05). (4) Compared with (1.2±0.9)% in 10.0% oxygen+ 10.0% FCS group, the cell apoptosis rate in 5.0% oxygen+ 5.0% FCS group showed no significant difference [(2.6±0.9)%, P >0.05], while it was significantly increased in 0.5% oxygen+ 0.5% FCS group [(13.3±4.1)%, P <0.05]. Conclusions: Severe hypoxia and low concentration of serum protein can inhibit proliferation activity and production of total collagen of human HSFs and induce their apoptosis.

A prospective randomized comparison of blood concentration as well as manifestations of ototoxicity and nephrotoxicity in groups with conventional and individualized administration of gentamicin was carried out in a total of 106 hospitalized patients. Therapeutic serum concentrations (peak 4-9 mg/L, though less than 1 mg/L) were achieved in 50% of the patients of conventional group and in all the patients of individualized group. Incidence of auditory and renal toxic reactions was high in the conventional group, being 27% and 30% respectively, while in the individualized group, the corresponding figures were 2.8% and 0%. Therefore, individualized administration of gentamicin based on data from therapeutic drug monitoring can markedly decrease the toxic effect and increase the therapeutic efficacy. It is worthwhile to use this kind of administration clinically.